Genes & Developmentgenesdev.cshlp.org/.../gad.270421.115.DC1/SuppMaterial.docx · Web...
Transcript of Genes & Developmentgenesdev.cshlp.org/.../gad.270421.115.DC1/SuppMaterial.docx · Web...
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SUPPLEMENTAL FIGURE LEGENDS
Supplemental Figure 1. D. melanogaster cell lines express circular RNAs from the PlexA
and laccase2 loci. (A) 20 micrograms of total RNA from DL1 and S2 cells was subjected to
Northern blot analysis and probed for PlexA expression. The locations of the probes are noted to
the right, with Probe 3 spanning the backsplicing junction. (B) RNA from DL1 cells was reverse
transcribed using random hexamers. PCR amplification was then performed using forward (5’-
CCTGCGCTATCTGCTCCTGA) and reverse (5’- GCTAGGATTGAGGATGGAGCTCC)
primers with genomic DNA (gDNA) or cDNA as a template. Products were then subjected to
Sanger sequencing. The backsplicing junction is denoted by the change in color from orange to
blue. (C) 20 micrograms of total RNA from DL1 and S2 cells was subjected to Northern blot
analysis and probed for Laccase2 expression. The locations of the Laccase2 Probe 1 and Probe 3
sequences are noted in Figure 1F. Probes to exon 3 and exon 4 confirmed the identity of the
Laccase2 mRNA transcript. -actin was used as a loading control.
Supplemental Figure 2. Intronic repeats facilitate Laccase2 circular RNA production in D.
melanogaster. (A) Sequence alignment of the upstream and downstream DNAREP1_DM
repeats, demonstrating that three distinct regions of the repeats are highly complementary to one
another. Numbering scheme is the same as that used in Figure 2A. (B) 8 micrograms of total
RNA from D. melanogaster (w1118 strain) and D. yakuba was subjected to Northern blot analysis
and probed for Laccase2 expression. 18S rRNA was used as a loading control. (C) As in Figure
1G, plasmids containing the laccase2 region in the sense or antisense orientations were
transfected into DL1 (left) or S2 cells (right). Northern blots were then performed using a probe
that spans the backsplicing junction.
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Supplemental Figure 3. Multiple hnRNPs and SR proteins regulate the expression of the
Laccase2 circular RNA. (A) Drosophila DL1 cells were bathed in the indicated dsRNAs for 3
days, and Northern blots performed to analyze the expression of the endogenous laccase2 locus.
For each gene, two independent, non-overlapping dsRNAs were tested (3 dsRNAs were used for
Mbl). Representative blots are shown. Circular RNA levels were quantified using ImageQuant
from three independent experiments and normalized to the No dsRNA samples. Data at the
bottom are shown as mean±SD. (B) qPCR confirmed that the key dsRNAs used in Figure 3B
each efficiently depleted their target gene, whereas -gal dsRNA had no significant effect. Data
from three independent experiments were normalized to -actin (Act42a) and are shown as
mean±SD. (C) Quantification of Northern blots from Figure 3D that examined Mbl circular
RNA expression in SL2 cells that had been treated with the indicated dsRNAs for 4 days.
Circular RNA levels were quantified using ImageQuant from three independent experiments and
normalized to the No dsRNA samples. Data are shown as mean±SD.
Supplemental Figure 4. hnRNP and SR proteins regulate the expression of other
Drosophila circular RNAs. Drosophila DL1 cells were bathed in the indicated dsRNAs for 3
days, and qPCR was used to examine the expression of nine endogenous circular RNAs. Primers
that span the backsplicing junction (and thus only detect the circular RNA, not the linear mRNA)
were used. Although the Pan circular RNA was largely unaffected by any of the dsRNAs, the
expression of the other circular RNAs was significantly affected by one or more of the tested
factors.
Supplemental Figure 5. The laccase2 introns support circularization of diverse exons in S2
cells. (A) The plasmids used in Figures 6D-F were transfected into S2 cells and Northern blots
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performed. As in DL1 cells, circular RNAs composed of the ZKSCAN1, HIPK3, or ciRS7 exons
were all efficiently generated. (B) Portions of the cGFP ORF were sequentially cloned into the
multicloning site of the Laccase2 MCS Vector, such that the complete ORF is present in a split
orientation (Top). The regions targeted by Northern oligonucleotide probes are denoted in red.
Plasmids were then transfected into DL1 cells and Northern blots performed (Bottom). A
properly backspliced split GFP circular RNA was efficiently produced.
Supplemental Figure 6. Efficient circular RNA production from plasmids in human cell
lines. (A) The 1945-nt region of the Laccase2 pre-mRNA was inserted in the sense or antisense
orientation into pcDNA3.1(+) and transfected into HeLa cells. Consistent with the results in
Drosophila DL1 and S2 cells, the Laccase2 sense plasmid efficiently generated a circular RNA
in HeLa cells. * represents a non-specific band that is also present in Mock treated cells. (B)
Exon 2 of human HIPK3 was inserted into the multicloning sites of the designated plasmids.
Plasmids were then transfected into HeLa cells (which naturally express the HIPK3 circular
RNA from the endogenous loci) and Northern blots performed. Whereas the CircRNA Mini
vector was unable to generate this circle, the presence of long flanking repeats (in either the
ZKSCAN1 MCS or Laccase2 MCS vectors) greatly improved circularization efficiency. Of
particular note, the Laccase2 MCS backbone generated only a minimal amount of linear RNA.
(C) Segments of human ZKSCAN1 exons 2 and 3 (numbering scheme from (Liang and Wilusz
2014)) were inserted into the multicloning site of the ZKSCAN1 MCS Vector. Plasmids were
then transfected into HeLa cells and Northern blots performed. Exons ≥500-nt circularized most
efficiently (e.g., 548-1047), although some circularization was detected using a 300-nt exon
(548-847). (D) The ZKSCAN1 MCS plasmids described in Figure 7C were transfected into
HeLa cells. Total RNA was then isolated and treated with RNase R. Whereas the linear
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MALAT1 transcript was efficiently degraded by RNase R (bottom), the ZKSCAN1 circular
RNAs derived from the plasmids were all resistant to degradation (top). (E) The ZKSCAN1
circular RNAs derived from the ZKSCAN1 MCS plasmids localized to the cytoplasm. A probe
to endogenous MALAT1 was used as a control for fractionation efficiency. (F) The indicated
regions of ZKSCAN1 were inserted into the ZKSCAN1 MCS vector (Figure 7B, bottom)
between the EcoRV-SacII or EcoRV-PacI restriction sites. Plasmids were then transfected into
HeLa cells and Northern blots performed. As expected, the EcoRV-PacI circular RNAs were 27-
nt longer than the EcoRV-SacII transcripts due to the extra restriction sites present at the 3’ end
of the exon. Nevertheless, each exon circularized at a similar efficiency regardless of the cloning
strategy.
Supplemental Figure 7. Circular RNAs consisting of an ORF and IRES likely can be
translated. (A) Portions of the cGFP ORF were sequentially cloned into the multicloning site of
the ZKSCAN1 MCS Vector. The 5’ end of the ORF (amino acids 1-173) was first inserted to
generate the “GFP 5’ Half Only” Vector (top). The 3’ end of the ORF (amino acids 174 to the
stop codon) was then inserted and Quikchange used to change the restriction sites to the wildtype
GFP sequence. As the 5’ and 3’ halves of the ORF were inserted in a split orientation,
backsplicing is required to generate the full-length ORF. The EMCV IRES was then inserted in
the sense or antisense orientation upstream of the cGFP start codon (bottom). (B) The indicated
plasmids were transfected into HeLa cells and Northern blots performed. Although some linear
RNAs were generated, the predominant transcripts produced from the plasmids were the circular
RNAs. (C) cGFP expression was detected by fluorescence microscopy. Few GFP positive cells
were observed when no IRES was present (WT Split GFP construct) or when the IRES was
present in the antisense orientation. In contrast, the WT Split GFP + Sense IRES plasmid yielded
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many GFP positive cells. pIRES2-EGFP served as a control for IRES activity. Representative
images are shown.
Supplemental Figure 8. Simple repeats facilitate circular RNA production from the
Drosophila Sema-2b gene. (A) Exon/intron structure of the D. melanogaster Semaphorin-2b
(Sema-2b) locus, highlighting a 980-nt region that includes exons 8 and 9. A circular RNA is
formed when the 5’ splice site at the end of exon 9 is joined to the 3’ splice site at the beginning
of exon 8 (purple). Repetitive elements in the designated orientations are shown (along with their
positions in the 980-nt cloned insert). The regions targeted by Northern oligonucleotide probes
are denoted in red. (B) The 980-nt region of the Sema-2b pre-mRNA was cloned downstream of
the metallothionein promoter to generate the “Sema-2b nt 1-980” plasmid. The plasmid was then
transfected into DL1 cells, and CuSO4 was added for 14 hr where indicated. Northern blots were
subsequently performed with three different oligonucleotide probes, confirming that the Sema-2b
circular RNA is produced. -actin was used as a loading control. (C) The plasmid-derived Sema-
2b circular RNA was resistant to RNase R digestion. (D) Sema-2b expression plasmids
containing deletions at their 5’ or 3’ ends were transfected into DL1 cells, and Northern blots
performed. Whereas the 1-930 plasmid contains both flanking repeat elements and generated the
circular RNA, efficient circularization was lost when either of the flanking simple repeat
sequences were deleted.
Supplemental Figure 9. The biogenesis of the endogenous Laccase2 circular RNA and a
plasmid-derived Laccase2 circular RNA are regulated differently. To determine whether the
various hnRNP and SR proteins may regulate Laccase2 circular RNA biogenesis or the stability
of the mature circular RNA, we generated a stable DL1 cell line expressing the Hy_pMT
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Laccase2 MCS-Laccase2 608-1097 plasmid (Figures 6A-B) using selection with Hygromycin.
As described in the main text, this plasmid lacks the flanking laccase2 exons as well as most of
the flanking introns. Compared to the endogenous Laccase2 circular RNA (490-nt), the mature
plasmid-derived Laccase2 circular RNA is 24-nt longer due to the presence of the KpnI and
XmaI/SacI sites at the ends of the exon (Top). The DL1 stable cell line was then treated with the
indicated dsRNAs for 3 days, and CuSO4 was added for the last 14 hr to induce transcription
from the plasmid’s metallothionein promoter. By using Northern probes against the unique
backspliced junctions, we were able to distinguish between the endogenous and plasmid-derived
Laccase2 circular RNAs. Whereas expression of the endogenous Laccase2 circular RNA
increased upon depletion of Hrb27C, SF2, SRp54, or B52, expression of the plasmid-derived
circular RNA either did not change (Hrb27C, SF2) or decreased (SRp54, B52). This is consistent
with a model in which these factors regulate Laccase2 backsplicing rather than the stability of
the mature circular RNA. Depletion of Hrb87F caused the expression of both the endogenous
and plasmid-derived circular RNAs to decrease, suggesting Hrb87F may regulate backsplicing
and/or circular RNA stability.
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SUPPLEMENTAL METHODS
All Drosophila expression plasmids were generated from the Hy_pMT EGFP SV40 pA Sense plasmid, which is a modified form of the pMK33/pMtHy plasmid. In brief, the metallothionein promoter (marked in blue) drives expression of the EGFP ORF (marked in green) that terminates in the SV40 polyadenylation signal (marked in pink). HygroB and Amp selectable markers are also present. The full plasmid sequence is as follows:
GATCAATTCGTTGCAGGACAGGATGTGGTGCCCGATGTGACTAGCTCTTTGCTGCAGGCCGTCCTATCCTCTGGTTCCGATAAGAGACCCAGAACTCCGGCCCCCCACCGCCCACCGCCACCCCCATACATATGTGGTACGCAAGTAAGAGTGCCTGCGCATGCCCCATGTGCCCCACCAAGAGCTTTGCATCCCATACAAGTCCCCAAAGTGGAGAACCGAACCAATTCTTCGCGGGCAGAACAAAAGCTTCTGCACACGTCTCCACTCGAATTTGGAGCCGGCCGGCGTGTGCAAAAGAGGTGAATCGAACGAAAGACCCGTGTGTAAAGCCGCGTTTCCAAAATGTATAAAACCGAGAGCATCTGGCCAATGTGCATCAGTTGTGGTCAGCAGCAAAATCAAGTGAATCATCTCAGTGCAACTAAAGGGGGGATCTCGAGGTCGACGGTATCGATAAGCTTGATATCACCATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCCTGACCTACGGCGTGCAGTGCTTCAGCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAACGTCTATATCATGGCCGACAAGCAGAAGAACGGCATCAAGGTGAACTTCAAGATCCGCCACAACATCGAGGACGGCAGCGTGCAGCTCGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCACCCAGTCCGCCCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTGTACAAGTAAGCGGCCGCAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTAACTAGTGGATCCACTAGGGGCCGCCGACGCGAGGCTGGATGGCCTTCCCCATTATGATTCTTCTCGCTTCCGGCGGCATCGGGATGCCCGCGTTGCAGGCCATGCTGTCCAGGCAGGTAGATGACGACCATCAGGGACAGCTTCAAGGATCGCTCGCGGCTCTTACCAGCCTAACTTCGATCATTGGACCGCTGATCGTCACGGCGATTTATGCCGCCTCGGCGAGCACATGGAACGGGTTGGCATGGATTGTAGGCGCCGCCCTATACCTTGTCTGCCTCCCCGCGTTGCGTCGCGGTGCATGGAGCCGGGCCACCTCGACCTGAATGGAAGCCGGCGGCACCTCGCTAACGGATTCACCACTCCAAGAATTGGAGCCAATCAATTCTTGCGGAGAACTGTGAATGCGCAAACCAACCCTTGGCAGAACATATCCATCGCGTCCGCCATCTCCAGCAGCCGCACGCGGCGCATCTCGGGCAGCGTTGGGTCCTGGCCACGGGTGCGCATGATCGTGCTCCTGTCGTTGAGGACCCGGCTAGGCTGGCGGGGTTGCCTTACTGGTTAGCAGAATGAATCACCGATACGCGAGCGAACGTGAAGCGACTGCTGCTGCAAAACGTCTGCGACCTGAGCAACAACATGAATGGTCTTCGGTTTCCGTGTTTCGTAAAGTCTGGAAACGCGGAAGTCAGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTGCAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAACACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTCTTCAAGAATTCACATTTGTACGAATTTTTTTTTTATCAAAAGTTCGAGTTTTTCACCAATTTCCTCATCAACCGAGCAAGGCAAACGGCTTTGAATAATATGGTGTTATATATACATATATCAAATCGCTGCTGACTGCGTGATTGATGGCCCCAAGATTACATATTATCGAATCAGGATTCAGAAGGAGATCAATGTCAAATGCGGACAGGAACATGAAAGACGCCTGTTATGCGCAATTAAAAATTTGGGTTTAATTGCTGTGGAAACTGTTGTTGGCGGCATCTTAAGTTCCTGTTTAACAACATCAACTACTTATGTACGTAGAAGCGTTTAAGCCATTTGCATACAGATGAGAACTGGCTTTTGTGCTAATCAGTCAAGATGACTCCGATGATGATGACTCATTACCTGACCAGTTTTCGCTGCTTTCTTTTCAACAACTACTTGTATATGTATTGTATCCAATAGCAATACATTGAATTTCCATGGTCTAGTCACGTATTATCATTTAATTGACACCAAGTCGTGTTATTGTTGAGCTATCGAGTTCAGCTCAAACATTTCTTATTCCCATGAATAAGCCGGCAAAAATATGCAATCTATGAAAGTTAATATAAGCAAACCTTACTTTGACTCAATACCAATGCACTTTGTGTCGATAGGTTCACGCAATTGAGGCGATTATTCCGATAACCCAAGCGATTGACTGTTCCCGTTTCGATTCCAATTGAAATTTGGAAATGTACAATAGTTTTGCTATATGCTGTCAAGTACGCTCTTATCTTCTCTGGGTTTTCTTCAGAGTTTCGAAACGCTTCTTCTTTTTTTTGTTTTTTTTTTTTTGGAATCTCGTATTTTGGAAGGGGCTCCCCTCTGGAATTTGTTACACTGTCGTTATCATTGCGAACAAGCGGCCCGAAGCTATCAGCGACTTTAACATTTACAATGCACTTTTTTACGACCAATTAAATGTACATTTTCCTTTCTTCGCCCGTTGATAAGCGAACGCGATGTGGCGCAGGCAATGTGTTGCTCTTGCGACACAAACGCAATCAAAATGGATTCAATTTCGCTTTTTCCCAGTGAAACGAAGAACGAACCGACCATCATGATATGCTCCTCTGCATGTTGCGTATTGAATCAATGACAATTTCAATTAAGCCGCCCGTTCGTCATGCGTTTTCGTGCGCTTCGAAATGCTGATAACGCTGCTGTCCTCCAACTGCTTTGCATGTGGACACAATTCCATTTATTTAATTCTTTTATTTGGATCGGTTAAATTAAAAAGCGCCTTGTTACGCATTTAACGTTGTTTCCGGTGCGTGGTGGTTTCATGCTTCTGGGAACGGCAAATGGGTTTAGGATTGGGAACCCCTCATCATCTGTTGGAATATACTATTCAACCTACAAAAGTAACGTTAAACAACACTACTTTATATTTGATATGAATGGCCACACCTTTTATGCCATAAAACATATTGTAAGAGAATACCACTCTTTTTATTCCTTCTTTCCTTCTTGTACGTTTTTTGCTGTAAGTAGGTCGTGGTGCTGGTGTTGCAGTTGAAATAACTTAAAATATAAATCATAAAACTCAAACATAAA
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CTTGACTATTTATTTATTTATTAAGAAAGGAAATATAAATTATAAATTACAACAGGTTATGGACCTGCAGCCAAGCTTGGCGCTCGTCCGGGGGCAATGAGATATGAAAAAGCCTGAACTCACCGCGACGTCTGTCGAGAAGTTTCTGATCGAAAAGTTCGACAGCGTCTCCGACCTGATGCAGCTCTCGGAGGGCGAAGAATCTCGTGCTTTCAGCTTCGATGTAGGAGGGCGTGGATATGTCCTGCGGGTAAATAGCTGCGCCGATGGTTTCTACAAAGATCGTTATGTTTATCGGCACTTTGCATCGGCCGCGCTCCCGATTCCGGAAGTGCTTGACATTGGGGAATTCAGCGAGAGCCTGACCTATTGCATCTCCCGCCGTGCACAGGGTGTCACGTTGCAAGACCTGCCTGAAACCGAACTGCCCGCTGTTCTGCAGCCGGTCGCGGAGGCCATGGATGCGATCGCTGCGGCCGATCTTAGCCAGACGAGCGGGTTCGGCCCATTCGGACCGCAAGGAATCGGTCAATACACTACATGGCGTGATTTCATATGCGCGATTGCTGATCCCCATGTGTATCACTGGCAAACTGTGATGGACGACACCGTCAGTGCGTCCGTCGCGCAGGCTCTCGATGAGCTGATGCTTTGGGCCGAGGACTGCCCCGAAGTCCGGCACCTCGTGCACGCGGATTTCGGCTCCAACAATGTCCTGACGGACAATGGCCGCATAACAGCGGTCATTGACTGGAGCGAGGCGATGTTCGGGGATTCCCAATACGAGGTCGCCAACATCTTCTTCTGGAGGCCGTGGTTGGCTTGTATGGAGCAGCAGACGCGCTACTTCGAGCGGAGGCATCCGGAGCTTGCAGGATCGCCGCGGCTCCGGGCGTATATGCTCCGCATTGGTCTTGACCAACTCTATCAGAGCTTGGTTGACGGCAATTTCGATGATGCAGCTTGGGCGCAGGGTCGATGCGACGCAATCGTCCGATCCGGAGCCGGGACTGTCGGGCGTACACAAATCGCCCGCAGAAGCGCGGCCGTCTGGACCGATGGCTGTGTAGAAGTACTCGCCGATAGTGGAAACCGACGCCCCAGCACTCGTCCGAGGGCAAAGGAATAGAGTAGATGCCGACCGAACAAGAGCTGATTTCGAGAACGCCTCAGCCAGCAACTCGCGCGAGCCTAGCAAGGCAAATGCGAGAGAACGGCCTTACGCTTGGTGGCACAGTTCTCGTCCACAGTTCGCTAAGCTCGCTCGGCTGGGTCGCGGGAGGGCCGGTCGCAGTGATTCAGGCCCTTCTGGATTGTGTTGGTCCCCAGGGCACGATTGTCATGCCCACGCACTCGGGTGATCTGACTGATCCCGCAGATTGGAGATCGCCGCCCGTGCCTGCCGATTGGGTGCAGATCTTTGTGAAGGAACCTTACTTCTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTAAGGTAAATATAAAATTTTTAAGTGTATAATGTGTTAAACTACTGATTCTAATTGTTTGTGTATTTTAGATTCCAACCTATGGAACTGATGAATGGGAGCAGTGGTGGAATGCCTTTAATGAGGAAAACCTGTTTTGCTCAGAAGAAATGCCATCTAGTGATGATGAGGCTACTGCTGACTCTCAACATTCTACTCCTCCAAAAAAGAAGAGAAAGGTAGAAGACCCCAAGGACTTTCCTTCAGAATTGCTAAGTTTTTTGAGTCATGCTGTGTTTAGTAATAGAACTCTTGCTTGCTTTGCTATTTACACCACAAAGGAAAAAGCTGCACTGCTATACAAGAAAATTATGGAAAAATATTCTGTAACCTTTATAAGTAGGCATAACAGTTATAATCATAACATACTGTTTTTTCTTACTCCACACAGGCATAGAGTGTCTGCTATTAATAACTATGCTCAAAAATTGTGTACCTTTAGCTTTTTAATTTGTAAAGGGGTTAATAAGGAATATTTGATGTATAGTGCCTTGACTAGAGATCATAATCAGCCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTG
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The following plasmids were generated by inserting the indicated sequences into the XhoI and NotI sites of Hy_pMT EGFP SV40 pA Sense, thereby replacing the EGFP ORF with the Laccase2 insert.
Hy_pMT Laccase2 Sense (Exon in yellow)Used in Figures 1G, 2B, 2C, 2E, 5BUsed in Supplemental Figure 2CCATTGAGAAATGACTGAGTTCCGGTGCTCTCAAGTCATTGATCTTTGTCGACTTTTATTTGGTCTCTGTAATAACGACTTCAAAAACATTAAATTCTGTTGCGAAGCCAGTAAGCTACAAAAAGAAAaaacaagagagaatgctatagtcgtatagtatagtttcccgactatctgatacccattacttatctagggggaatgcgaacccaaaattttatcagttttctcggatatcgatagatattggggaataaatttaaataaataaattttgggcgggtttagggcgtggcaaaaagttttttggcaaatcgctagaaatttacaagacttataaaattatgaaaaaatacaacaaaattttaaacacgtgggcgtgacagttttggacggttttagggcgttagagtaggcgaggacagggttacatcgactaggctttgatcctgatcaagaatatatatactttataccgcttccttctacatgttacctatttttcaacgaatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagattcgttgaaaagtatgtaacaggaagaataaagcatttccgaccatgtaaagtatatatattcttaataaggatcaatagccgagtcgatctcgccatgtccgtctgtcttattattttattaccgccgagacatcaggaactataaaagctagaaggatgagttttagcatacagattctagagacaaggacgcagagcaagtttgttgatccatgctgccacgctttaactttctcaaattgcccaaaactgccatgcccacatttttgaactattttcgaaattttttcataattgtattactcgtgtaaatttccatcaatttgccaaaaaactttttgtcacgcgttaacgccctaaagccgccaatttggtcacgcccacactattgaacaattatcaaattttttctcattttattccccaatatctatcgatatccccgattatgaaattattaaatttcgcgttcgcattcacactagctgagtaacgagtatctgatagttggggaaatcgactTATTTTTTATATACAATGAAAATGAATTTAATCATATGAATATCGATTATAGCTTTTTATTTAATATGAATATTTATTTGGGCTTAAGGTGTAACCTcctcgacataagactcacatggcgcaggcacattgaagacaaaaatactcaTTGTCGGGTCTCGCACCCTCCAGCAGCACCTAAAATTATGTCTTCAATTATTGCCAACATTGGAGACACAATTAGTCTGTGGCACCTCAG
Hy_pMT Laccase2 AntisenseUsed in Figure 1GUsed in Supplemental Figure 2CCTGAGGTGCCACAGACTAATTGTGTCTCCAATGTTGGCAATAATTGAAGACATAATTTTAGGTGCTGCTGGAGGGTGCGAGACCCGACAAtgagtatttttgtcttcaatgtgcctgcgccatgtgagtcttatgtcgaggAGGTTACACCTTAAGCCCAAATAAATATTCATATTAAATAAAAAGCTATAATCGATATTCATATGATTAAATTCATTTTCATTGTATATAAAAAATAagtcgatttccccaactatcagatactcgttactcagctagtgtgaatgcgaacgcgaaatttaataatttcataatcggggatatcgatagatattggggaataaaatgagaaaaaatttgataattgttcaatagtgtgggcgtgaccaaattggcggctttagggcgttaacgcgtgacaaaaagttttttggcaaattgatggaaatttacacgagtaatacaattatgaaaaaatttcgaaaatagttcaaaaatgtgggcatggcagttttgggcaatttgagaaagttaaagcgtggcagcatggatcaacaaacttgctctgcgtccttgtctctagaatctgtatgctaaaactcatccttctagcttttatagttcctgatgtctcggcggtaataaaataataagacagacggacatggcgagatcgactcggctattgatccttattaagaatatatatactttacatggtcggaaatgctttattcttcctgttacatacttttcaacgaatctaCAATAGTATACAATAGTATAGAAACTGCCTATTAAAAAATCGAATATTTCTAGTAAAAAATTTTGGAATTTTGAATACTTACGCTCCCAATACTGTGTAGTATTCCAACGTAAAATGATAGTAGCAAATTCTCGGAGGCTCGCCTTCGCGACAAGCGCGTGCACATTCATCAGGAGCAGATAGCGCAGGATTGCGCCGCAGCTCGGCAGTTGCACTAGTAGAGAAATCTAAGTGGCGGAATGGAGATTTTGGGTTGGGTTCCACGACGGTTCCTGTGATCTTATTAGCTGGATAACGTGCAGAGGTCGGTAGACTCCCGCGTGAGTTTAGGTAACCGCTTCCAGCGGTTAGGGCTGGCTTAGTACCAGCTAGGATTGAGGATGGAGCTCCAGACCCAAGGCTATTGCCATTTAATGGAGCGGGCCCAGAGTGGCTTCCACCTATAGACGGGTGAGTCTGCACAAGGCCGTGTGTTGCCGAAAAAAATGGTGAATCGAATGGAGTTGAACTACTAGGAGTGGCCACATGTTGCTGCCAGAAGGACCCCATTGGTGTAAGATCTGGTTGATTTTCTGCataaaataaaaaaaatatatataacaataaaaCTTAGTCTCGATTTAGCTTATTAttatacccataaatcgtacagtaaaaaggtatactagattcgttgaaaaataggtaacatgtagaaggaagcggtataaagtatatatattcttgatcaggatcaaagcctagtcgatgtaaccctgtcctcgcctactctaacgccctaaaaccgtccaaaactgtcacgcccacgtgtttaaaattttgttgtattttttcataattttataagtcttgtaaatttctagcgatttgccaaaaaactttttgccacgccctaaacccgcccaaaatttatttatttaaatttattccccaatatctatcgatatccgagaaaactgataaaattttgggttcgcattccccctagataagtaatgggtatcagatagtcgggaaactatactatacgactatagcattctctcttgtttTTTCTTTTTGTAGCTTACTGGCTTCGCAACAGAATTTAATGTTTTTGAAGTCGTTATTACAGAGACCAAATAAAAGTCGACAAAGATCAATGACTTGAGAGCACCGGAACTCAGTCATTTCTCAATG
Hy_pMT Laccase2 200-1945Used in Figure 2BaatgcgaacccaaaattttatcagttttctcggatatcgatagatattggggaataaatttaaataaataaattttgggcgggtttagggcgtggcaaaaagttttttggcaaatcgctagaaatttacaagacttataaaattatgaaaaaatacaacaaaattttaaacacgtgggcgtgacagttttggacggttttagggcgttagagtaggcgaggacagggttacatcgactaggctttgatcctgatcaagaatatatatactttataccgcttccttctacatgttacctatttttcaacgaatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCC
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GAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagattcgttgaaaagtatgtaacaggaagaataaagcatttccgaccatgtaaagtatatatattcttaataaggatcaatagccgagtcgatctcgccatgtccgtctgtcttattattttattaccgccgagacatcaggaactataaaagctagaaggatgagttttagcatacagattctagagacaaggacgcagagcaagtttgttgatccatgctgccacgctttaactttctcaaattgcccaaaactgccatgcccacatttttgaactattttcgaaattttttcataattgtattactcgtgtaaatttccatcaatttgccaaaaaactttttgtcacgcgttaacgccctaaagccgccaatttggtcacgcccacactattgaacaattatcaaattttttctcattttattccccaatatctatcgatatccccgattatgaaattattaaatttcgcgttcgcattcacactagctgagtaacgagtatctgatagttggggaaatcgactTATTTTTTATATACAATGAAAATGAATTTAATCATATGAATATCGATTATAGCTTTTTATTTAATATGAATATTTATTTGGGCTTAAGGTGTAACCTcctcgacataagactcacatggcgcaggcacattgaagacaaaaatactcaTTGTCGGGTCTCGCACCCTCCAGCAGCACCTAAAATTATGTCTTCAATTATTGCCAACATTGGAGACACAATTAGTCTGTGGCACCTCAG
Hy_pMT Laccase2 300-1945Used in Figure 2BagttttttggcaaatcgctagaaatttacaagacttataaaattatgaaaaaatacaacaaaattttaaacacgtgggcgtgacagttttggacggttttagggcgttagagtaggcgaggacagggttacatcgactaggctttgatcctgatcaagaatatatatactttataccgcttccttctacatgttacctatttttcaacgaatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagattcgttgaaaagtatgtaacaggaagaataaagcatttccgaccatgtaaagtatatatattcttaataaggatcaatagccgagtcgatctcgccatgtccgtctgtcttattattttattaccgccgagacatcaggaactataaaagctagaaggatgagttttagcatacagattctagagacaaggacgcagagcaagtttgttgatccatgctgccacgctttaactttctcaaattgcccaaaactgccatgcccacatttttgaactattttcgaaattttttcataattgtattactcgtgtaaatttccatcaatttgccaaaaaactttttgtcacgcgttaacgccctaaagccgccaatttggtcacgcccacactattgaacaattatcaaattttttctcattttattccccaatatctatcgatatccccgattatgaaattattaaatttcgcgttcgcattcacactagctgagtaacgagtatctgatagttggggaaatcgactTATTTTTTATATACAATGAAAATGAATTTAATCATATGAATATCGATTATAGCTTTTTATTTAATATGAATATTTATTTGGGCTTAAGGTGTAACCTcctcgacataagactcacatggcgcaggcacattgaagacaaaaatactcaTTGTCGGGTCTCGCACCCTCCAGCAGCACCTAAAATTATGTCTTCAATTATTGCCAACATTGGAGACACAATTAGTCTGTGGCACCTCAG
Hy_pMT Laccase2 400-1945Used in Figure 2BagggcgttagagtaggcgaggacagggttacatcgactaggctttgatcctgatcaagaatatatatactttataccgcttccttctacatgttacctatttttcaacgaatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagattcgttgaaaagtatgtaacaggaagaataaagcatttccgaccatgtaaagtatatatattcttaataaggatcaatagccgagtcgatctcgccatgtccgtctgtcttattattttattaccgccgagacatcaggaactataaaagctagaaggatgagttttagcatacagattctagagacaaggacgcagagcaagtttgttgatccatgctgccacgctttaactttctcaaattgcccaaaactgccatgcccacatttttgaactattttcgaaattttttcataattgtattactcgtgtaaatttccatcaatttgccaaaaaactttttgtcacgcgttaacgccctaaagccgccaatttggtcacgcccacactattgaacaattatcaaattttttctcattttattccccaatatctatcgatatccccgattatgaaattattaaatttcgcgttcgcattcacactagctgagtaacgagtatctgatagttggggaaatcgactTATTTTTTATATACAATGAAAATGAATTTAATCATATGAATATCGATTATAGCTTTTTATTTAATATGAATATTTATTTGGGCTTAAGGTGTAACCTcctcgacataagactcacatggcgcaggcacattgaagacaaaaatactcaTTGTCGGGTCTCGCACCCTCCAGCAGCACCTAAAATTATGTCTTCAATTATTGCCAACATTGGAGACACAATTAGTCTGTGGCACCTCAG
Hy_pMT Laccase2 450-1945Used in Figure 2BtgatcaagaatatatatactttataccgcttccttctacatgttacctatttttcaacgaatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagattcgttgaaaagtatgtaacaggaagaataaagcatttccgaccatgtaaagtatatatattcttaataaggatcaatagccgagtcgatctcgccatgtccgtctgtcttattattttattaccgccgagacatcaggaactataaaagctagaaggatgagttttagcatacagattctagagacaaggacgcagagcaagtttgttgatccatgctgccacgctttaactttctcaaattgcccaaaactgccatgcccacatttttgaactattttcgaaattttttcataattgtattactcgtgtaaatttccatcaatttgccaaaaaactttttgtcacgcgttaacgccctaaagccgccaatttggtcacgcccacactattgaacaattatcaaattttttctcattttattccccaatatctatcgatatccccgattatgaaattattaaatttcgcgttcgcattcacactagctgagtaacgagtatctgatagttggggaaatcgactTATTTTTTATATACAATGAAAATGAATTTAATCATATGAATATCGATTATAGCTTTTTATTTAATATGAATATTTATTTGGGCTTAAGGTGTAACCTcct
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cgacataagactcacatggcgcaggcacattgaagacaaaaatactcaTTGTCGGGTCTCGCACCCTCCAGCAGCACCTAAAATTATGTCTTCAATTATTGCCAACATTGGAGACACAATTAGTCTGTGGCACCTCAG
Hy_pMT Laccase2 500-1945Used in Figure 2BttttcaacgaatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagattcgttgaaaagtatgtaacaggaagaataaagcatttccgaccatgtaaagtatatatattcttaataaggatcaatagccgagtcgatctcgccatgtccgtctgtcttattattttattaccgccgagacatcaggaactataaaagctagaaggatgagttttagcatacagattctagagacaaggacgcagagcaagtttgttgatccatgctgccacgctttaactttctcaaattgcccaaaactgccatgcccacatttttgaactattttcgaaattttttcataattgtattactcgtgtaaatttccatcaatttgccaaaaaactttttgtcacgcgttaacgccctaaagccgccaatttggtcacgcccacactattgaacaattatcaaattttttctcattttattccccaatatctatcgatatccccgattatgaaattattaaatttcgcgttcgcattcacactagctgagtaacgagtatctgatagttggggaaatcgactTATTTTTTATATACAATGAAAATGAATTTAATCATATGAATATCGATTATAGCTTTTTATTTAATATGAATATTTATTTGGGCTTAAGGTGTAACCTcctcgacataagactcacatggcgcaggcacattgaagacaaaaatactcaTTGTCGGGTCTCGCACCCTCCAGCAGCACCTAAAATTATGTCTTCAATTATTGCCAACATTGGAGACACAATTAGTCTGTGGCACCTCAG
Hy_pMT Laccase2 1-1545Used in Figure 2CCATTGAGAAATGACTGAGTTCCGGTGCTCTCAAGTCATTGATCTTTGTCGACTTTTATTTGGTCTCTGTAATAACGACTTCAAAAACATTAAATTCTGTTGCGAAGCCAGTAAGCTACAAAAAGAAAaaacaagagagaatgctatagtcgtatagtatagtttcccgactatctgatacccattacttatctagggggaatgcgaacccaaaattttatcagttttctcggatatcgatagatattggggaataaatttaaataaataaattttgggcgggtttagggcgtggcaaaaagttttttggcaaatcgctagaaatttacaagacttataaaattatgaaaaaatacaacaaaattttaaacacgtgggcgtgacagttttggacggttttagggcgttagagtaggcgaggacagggttacatcgactaggctttgatcctgatcaagaatatatatactttataccgcttccttctacatgttacctatttttcaacgaatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagattcgttgaaaagtatgtaacaggaagaataaagcatttccgaccatgtaaagtatatatattcttaataaggatcaatagccgagtcgatctcgccatgtccgtctgtcttattattttattaccgccgagacatcaggaactataaaagctagaaggatgagttttagcatacagattctagagacaaggacgcagagcaagtttgttgatccatgctgccacgctttaactttctcaaattgcccaaaactgccatgcccacatttttgaactattttcgaaattttttcataattgtattactcgtgtaaatttccatcaatttgccaaaaaactttttgtcacgcgttaacgccctaa
Hy_pMT Laccase2 1-1445Used in Figure 2CCATTGAGAAATGACTGAGTTCCGGTGCTCTCAAGTCATTGATCTTTGTCGACTTTTATTTGGTCTCTGTAATAACGACTTCAAAAACATTAAATTCTGTTGCGAAGCCAGTAAGCTACAAAAAGAAAaaacaagagagaatgctatagtcgtatagtatagtttcccgactatctgatacccattacttatctagggggaatgcgaacccaaaattttatcagttttctcggatatcgatagatattggggaataaatttaaataaataaattttgggcgggtttagggcgtggcaaaaagttttttggcaaatcgctagaaatttacaagacttataaaattatgaaaaaatacaacaaaattttaaacacgtgggcgtgacagttttggacggttttagggcgttagagtaggcgaggacagggttacatcgactaggctttgatcctgatcaagaatatatatactttataccgcttccttctacatgttacctatttttcaacgaatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagattcgttgaaaagtatgtaacaggaagaataaagcatttccgaccatgtaaagtatatatattcttaataaggatcaatagccgagtcgatctcgccatgtccgtctgtcttattattttattaccgccgagacatcaggaactataaaagctagaaggatgagttttagcatacagattctagagacaaggacgcagagcaagtttgttgatccatgctgccacgctttaactttctcaaattgcccaaaactgccatgccc
Hy_pMT Laccase2 1-1345Used in Figure 2CCATTGAGAAATGACTGAGTTCCGGTGCTCTCAAGTCATTGATCTTTGTCGACTTTTATTTGGTCTCTGTAATAACGACTTCAAAAACATTAAATTCTGTTGCGAAGCCAGTAAGCTACAAAAAGAAAaaacaagagagaatgctatagtcgtatagtatagtttcccgactatctgatacccattacttatctagggggaatgcgaacccaaaattttatcagttttctcggatatcgatagatattggggaataaatttaaataaataaattttgggcgggtttagggcgtggcaaaaagttttttggcaaatcgctagaaatttacaagacttataaaattatgaaaaaatacaacaaaattttaaacacgtgggcgtgacagttttggacggttttagggcgttagagtaggcgaggacagggttacatcgactaggctttgatcctgatcaagaatatatatactttataccgcttccttc
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tacatgttacctatttttcaacgaatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagattcgttgaaaagtatgtaacaggaagaataaagcatttccgaccatgtaaagtatatatattcttaataaggatcaatagccgagtcgatctcgccatgtccgtctgtcttattattttattaccgccgagacatcaggaactataaaagctagaaggatga
Hy_pMT Laccase2 1-1245Used in Figure 2CCATTGAGAAATGACTGAGTTCCGGTGCTCTCAAGTCATTGATCTTTGTCGACTTTTATTTGGTCTCTGTAATAACGACTTCAAAAACATTAAATTCTGTTGCGAAGCCAGTAAGCTACAAAAAGAAAaaacaagagagaatgctatagtcgtatagtatagtttcccgactatctgatacccattacttatctagggggaatgcgaacccaaaattttatcagttttctcggatatcgatagatattggggaataaatttaaataaataaattttgggcgggtttagggcgtggcaaaaagttttttggcaaatcgctagaaatttacaagacttataaaattatgaaaaaatacaacaaaattttaaacacgtgggcgtgacagttttggacggttttagggcgttagagtaggcgaggacagggttacatcgactaggctttgatcctgatcaagaatatatatactttataccgcttccttctacatgttacctatttttcaacgaatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagattcgttgaaaagtatgtaacaggaagaataaagcatttccgaccatgtaaagtatatatatt
Hy_pMT Laccase2 1-1235Used in Figure 2CCATTGAGAAATGACTGAGTTCCGGTGCTCTCAAGTCATTGATCTTTGTCGACTTTTATTTGGTCTCTGTAATAACGACTTCAAAAACATTAAATTCTGTTGCGAAGCCAGTAAGCTACAAAAAGAAAaaacaagagagaatgctatagtcgtatagtatagtttcccgactatctgatacccattacttatctagggggaatgcgaacccaaaattttatcagttttctcggatatcgatagatattggggaataaatttaaataaataaattttgggcgggtttagggcgtggcaaaaagttttttggcaaatcgctagaaatttacaagacttataaaattatgaaaaaatacaacaaaattttaaacacgtgggcgtgacagttttggacggttttagggcgttagagtaggcgaggacagggttacatcgactaggctttgatcctgatcaagaatatatatactttataccgcttccttctacatgttacctatttttcaacgaatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagattcgttgaaaagtatgtaacaggaagaataaagcatttccgaccatgtaaag
Hy_pMT Laccase2 1-1225Used in Figure 2CCATTGAGAAATGACTGAGTTCCGGTGCTCTCAAGTCATTGATCTTTGTCGACTTTTATTTGGTCTCTGTAATAACGACTTCAAAAACATTAAATTCTGTTGCGAAGCCAGTAAGCTACAAAAAGAAAaaacaagagagaatgctatagtcgtatagtatagtttcccgactatctgatacccattacttatctagggggaatgcgaacccaaaattttatcagttttctcggatatcgatagatattggggaataaatttaaataaataaattttgggcgggtttagggcgtggcaaaaagttttttggcaaatcgctagaaatttacaagacttataaaattatgaaaaaatacaacaaaattttaaacacgtgggcgtgacagttttggacggttttagggcgttagagtaggcgaggacagggttacatcgactaggctttgatcctgatcaagaatatatatactttataccgcttccttctacatgttacctatttttcaacgaatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagattcgttgaaaagtatgtaacaggaagaataaagcatttccga
Hy_pMT Laccase2 1-1219Used in Figure 2CCATTGAGAAATGACTGAGTTCCGGTGCTCTCAAGTCATTGATCTTTGTCGACTTTTATTTGGTCTCTGTAATAACGACTTCAAAAACATTAAATTCTGTTGCGAAGCCAGTAAGCTACAAAAAGAAAaaacaagagagaatgctatagtcgtatagtatagtttcccgactatctgatacccattacttatctagggggaatgcgaacccaaaattttatcagttttctcggatatcgatagatattggggaataaatttaaataaataaattttgggcgggtttagggcgtggcaaaaagttttttggcaaatcgctagaaatttacaagacttataaaattatgaaaaaatacaacaaaattttaaacacgtgggcgtgacagttttggacggttttagggcgttagagtaggcgaggacagggttacatcgactaggctttgatcctgatcaagaatatatatactttataccgcttccttc
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tacatgttacctatttttcaacgaatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagattcgttgaaaagtatgtaacaggaagaataaagcat
Hy_pMT Laccase2 450-1245 Used in Figure 2EtgatcaagaatatatatactttataccgcttccttctacatgttacctatttttcaacgaatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagattcgttgaaaagtatgtaacaggaagaataaagcatttccgaccatgtaaagtatatatatt
Hy_pMT Laccase2 450-1245 Mut 5’ RepeatUsed in Figure 2EtgatcaagaatatatatactttataccgcttccttctacatgttacctatttttGTTGCTatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagattcgttgaaaagtatgtaacaggaagaataaagcatttccgaccatgtaaagtatatatatt
Hy_pMT Laccase2 450-1245 Mut 3’ RepeatUsed in Figure 2E tgatcaagaatatatatactttataccgcttccttctacatgttacctatttttcaacgaatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagatAGCAACaaaagtatgtaacaggaagaataaagcatttccgaccatgtaaagtatatatatt
Hy_pMT Laccase2 450-1245 Mut 5’+3’ RepeatUsed in Figure 2EtgatcaagaatatatatactttataccgcttccttctacatgttacctatttttGTTGCTatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagatAGCAACaaaagtatgtaacaggaagaataaagcatttccgaccatgtaaagtatatatatt
Hy_pMT Laccase2 MCS Exon Vector (Exon in yellow)Used in Figures 6B, 6C, 6D, 6E, 6F Used in Supplemental Figure 5ACATTGAGAAATGACTGAGTTCCGGTGCTCTCAAGTCATTGATCTTTGTCGACTTTTATTTGGTCTCTGTAATAACGACTTCAAAAACATTAAATTCTGTTGCGAAGCCAGTAAGCTACAAAAAGAAAaaacaagagagaatgctatagtcgtatagtatagtttcccgactatctgatacccattacttatctagggggaatgcgaacccaaaattttatcagttttctcggatatcgatagatattggggaataaatttaaataaataaattttgggcgggtttagggcg
Page 14
tggcaaaaagttttttggcaaatcgctagaaatttacaagacttataaaattatgaaaaaatacaacaaaattttaaacacgtgggcgtgacagttttggacggttttagggcgttagagtaggcgaggacagggttacatcgactaggctttgatcctgatcaagaatatatatactttataccgcttccttctacatgttacctatttttcaacgaatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAAGGTACCACCGGTTTAATTAAGGTAACCATGCATGCTAGCCCCGGGGAGCTCAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagattcgttgaaaagtatgtaacaggaagaataaagcatttccgaccatgtaaagtatatatattcttaataaggatcaatagccgagtcgatctcgccatgtccgtctgtcttattattttattaccgccgagacatcaggaactataaaagctagaaggatgagttttagcatacagattctagagacaaggacgcagagcaagtttgttgatccatgctgccacgctttaactttctcaaattgcccaaaactgccatgcccacatttttgaactattttcgaaattttttcataattgtattactcgtgtaaatttccatcaatttgccaaaaaactttttgtcacgcgttaacgccctaaagccgccaatttggtcacgcccacactattgaacaattatcaaattttttctcattttattccccaatatctatcgatatccccgattatgaaattattaaatttcgcgttcgcattcacactagctgagtaacgagtatctgatagttggggaaatcgactTATTTTTTATATACAATGAAAATGAATTTAATCATATGAATATCGATTATAGCTTTTTATTTAATATGAATATTTATTTGGGCTTAAGGTGTAACCTcctcgacataagactcacatggcgcaggcacattgaagacaaaaatactcaTTGTCGGGTCTCGCACCCTCCAGCAGCACCTAAAATTATGTCTTCAATTATTGCCAACATTGGAGACACAATTAGTCTGTGGCACCTCAG
Hy_pMT Laccase2 MCS-WT Split GFP (Exon in yellow)Used in Supplemental Figure 5BgaattcCATTGAGAAATGACTGAGTTCCGGTGCTCTCAAGTCATTGATCTTTGTCGACTTTTATTTGGTCTCTGTAATAACGACTTCAAAAACATTAAATTCTGTTGCGAAGCCAGTAAGCTACAAAAAGAAAaaacaagagagaatgctatagtcgtatagtatagtttcccgactatctgatacccattacttatctagggggaatgcgaacccaaaattttatcagttttctcggatatcgatagatattggggaataaatttaaataaataaattttgggcgggtttagggcgtggcaaaaagttttttggcaaatcgctagaaatttacaagacttataaaattatgaaaaaatacaacaaaattttaaacacgtgggcgtgacagttttggacggttttagggcgttagagtaggcgaggacagggttacatcgactaggctttgatcctgatcaagaatatatatactttataccgcttccttctacatgttacctatttttcaacgaatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGtttAaACgttatatatattttttttattttatGCAGGATggcagcgtgcagctggccgaccactaccagcagaatacccccatcggcgatggccctgtgctgctgcccgataaccactacctgtccacccagagcgccctgtccaaggaccccaacgagaagcgcgatcacatgatCTACTTCGGCttcgtgaccgccgccggcatcacccacggcatggacgagctgtacaagtgaATCGATACCGGTTTAATTAACACGTGGGTAACCGTTCCGAGTCGTTAACCACCatggccagcaagggcgCCgagctgttcaccggcAtCgtgcccatcctgAtCgagctgAaTggcgatgtgaatggccacaagttcagcgtgagcggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcctgtgccctggcccaccctggtgaccaccctgagctacggcgtgcagtgcttctcacgctaccccgatcacatgaagcagcacgacttcttcaagagcgccatgcctgagggctacatccaggagcgcaccatcttcttcgaggatgacggcaactacaagtcgcgcgccgaggtgaagttcgagggcgataccctggtgaatcgcatcgagctgaccggcaccgatttcaaggaggatggcaacatcctgggcaataagatggagtacaactacaacgcccacaatgtgtacatcatgaccgacaaggccaagaatggcatcaaggtgaacttcaagatccgccacaACATCGAGGTAAGTATTCAAAATTCCAAAATTTTTTCCTAGGAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagattcgttgaaaagtatgtaacaggaagaataaagcatttccgaccatgtaaagtatatatattcttaataaggatcaatagccgagtcgatctcgccatgtccgtctgtcttattattttattaccgccgagacatcaggaactataaaagctagaaggatgagttttagcatacagattctagagacaaggacgcagagcaagtttgttgatccatgctgccacgctttaactttctcaaattgcccaaaactgccatgcccacatttttgaactattttcgaaattttttcataattgtattactcgtgtaaatttccatcaatttgccaaaaaactttttgtcacgcgttaacgccctaaagccgccaatttggtcacgcccacactattgaacaattatcaaattttttctcattttattccccaatatctatcgatatccccgattatgaaattattaaatttcgcgttcgcattcacactagctgagtaacgagtatctgatagttggggaaatcgactTATTTTTTATATACAATGAAAATGAATTTAATCATATGAATATCGATTATAGCTTTTTATTTAATATGAATATTTATTTGGGCTTAAGGTGTAACCTcctcgacataagactcacatggcgcaggcacattgaagacaaaaatactcaTTGTCGGGTCTCGCACCCTCCAGCAGCACCTAAAATTATGTCTTCAATTATTGCCAACATTGGAGACACAATTAGTCTGTGGCACCTCAG
Hy_pMT Sema-2b nt 1-980 (Exons in yellow)Used in Supplemental Figures 8B, 8C, 8DGAGCACAAGGCACAAGcacacacacatacacacacacacatgcacacatacacaTGAGCAGATGAGTTCACTCAGAGAGGGAGGCACGTGCGTACACTTTATGCGTAATTGTGTGCGTGCGTGCCCTGGCTTACTGGGCGTATGCGTGATGTTCGAGCCTTATCAATAGTTGCCTCCTTGCTAACAATGTCTGTGTCATTTCTCTCTTTCCCCCTTCCCGTTAACCAAAAATGCAGTACGCCGGCACAAATGCCGAGTTCACCAAAGCGGATTCCGTCATATTCCGCTCGGACCTGTACAACCTGACGAATGGACGCAAGGAGGCCAACTTCAAGAGAACCGTGAAGTACGACTCCAAGCTGCTGGACAGTAAGTAGTGGCCATAAATCATATATTGCTAAGACAACTTTTAATGATTACATTTCTATTCATGGAAATGTGTAAAATCCAAATAATATGTAACAAATCACACATCAATCGCTTTTCTTTTCAGAGCCCAACTTCGTCGGCTCCTTTGAGATTGGTGAATTTGTGTACTTCTTCTTCCGCGAACATGCCGTTGAGTACATAAATTGCGGAAAAGCCGTCTACTCCCGAGTGGCTCGGGTGTGCAAAAACGACCGCGGTGGAAAATACATGATCAGCCAGAATTGGGCCACGTATCTGAAGGCGCGGATGAACTGCAGCATCTCCAGCGAATTTCCCTTCTACTTCAATGAGATACAATCCGTCTACAAGATGCCAACGGACGATACCAAGTTCTATGCCACGTTCACAACGAATACGAATGGTCTGATTGGATCGGCAGTGTGCAGCTACGACATCCGGGACATCAATGCGGCCTTCGACGGTGAGTATTCACTGTAGATATAGGTCGgtgtctgtgtgagtgtgtgtgtgtgtgtgtTGGCGGGAGGGTGTGGGATGCTCGTGTGATTTATTACAGTGTCCATAATTCTTCGGGGGGATTCCTTTTCTCC
Hy_pMT Sema-2b nt 50-980Used in Supplemental Figure 8DacacaTGAGCAGATGAGTTCACTCAGAGAGGGAGGCACGTGCGTACACTTTATGCGTAATTGTGTGCGTGCGTGCCCTGGCTTACTGGGCGTATGCGTGATGTTCGAGCCTTATCAATAGTTGCCTCCTTGCTAACAATGTCTGTGTCATTTCTCTCTTTCCCCCTTCCCGTTAACCAAAAATGCAGTACGCCGGCACAAATGCCGAGTTCACCAAAGCGGATTCCGTCATATTCCGCTCGGACCTGTACAACCTGACGAATGGACGCAAGGAGGCCAACTTCAAGAGAACCGTGAAGTACGACTCCAAGCTGCTGGACAGTAAGTAGTGGCCATAAATCATATATTGCTAAGACAACTTTTAATGATTACATTTCTATTCATGGAAATGTGTAAAATCCAAATAATATGTAACAAATCACACATCAATCGCTTTTCTTTTCAGAGCCCAACTTCGTCGGCTCCTTTGAGATTGGTGAATTTGTGTACTTCTTCTTCCGCGAACATGCCGTTGAGTACATAAATTGCGGAAAAGCCGTCTACTCCCGAGTGGCTCGGGTGTGCAAAAACGACCGCGGTGGAAAATACATGATCAGCCAGAATTGGGCCACGTATCTGAAGGCGCGGATGAACTGCAGCATCTCCAGCGAATTTCCCTTCTACTTCAATGAGATACAATCCGTCTACAAGATGCCAACGGACGATACCAAGTTCTATGCCACGTTCACAACGAATACGAATGGTCTGATTGGATCGGCAGTGTGCAGCTACGACATC
Page 15
CGGGACATCAATGCGGCCTTCGACGGTGAGTATTCACTGTAGATATAGGTCGgtgtctgtgtgagtgtgtgtgtgtgtgtgtTGGCGGGAGGGTGTGGGATGCTCGTGTGATTTATTACAGTGTCCATAATTCTTCGGGGGGATTCCTTTTCTCC
Hy_pMT Sema-2b nt 100-980Used in Supplemental Figure 8DTATGCGTAATTGTGTGCGTGCGTGCCCTGGCTTACTGGGCGTATGCGTGATGTTCGAGCCTTATCAATAGTTGCCTCCTTGCTAACAATGTCTGTGTCATTTCTCTCTTTCCCCCTTCCCGTTAACCAAAAATGCAGTACGCCGGCACAAATGCCGAGTTCACCAAAGCGGATTCCGTCATATTCCGCTCGGACCTGTACAACCTGACGAATGGACGCAAGGAGGCCAACTTCAAGAGAACCGTGAAGTACGACTCCAAGCTGCTGGACAGTAAGTAGTGGCCATAAATCATATATTGCTAAGACAACTTTTAATGATTACATTTCTATTCATGGAAATGTGTAAAATCCAAATAATATGTAACAAATCACACATCAATCGCTTTTCTTTTCAGAGCCCAACTTCGTCGGCTCCTTTGAGATTGGTGAATTTGTGTACTTCTTCTTCCGCGAACATGCCGTTGAGTACATAAATTGCGGAAAAGCCGTCTACTCCCGAGTGGCTCGGGTGTGCAAAAACGACCGCGGTGGAAAATACATGATCAGCCAGAATTGGGCCACGTATCTGAAGGCGCGGATGAACTGCAGCATCTCCAGCGAATTTCCCTTCTACTTCAATGAGATACAATCCGTCTACAAGATGCCAACGGACGATACCAAGTTCTATGCCACGTTCACAACGAATACGAATGGTCTGATTGGATCGGCAGTGTGCAGCTACGACATCCGGGACATCAATGCGGCCTTCGACGGTGAGTATTCACTGTAGATATAGGTCGgtgtctgtgtgagtgtgtgtgtgtgtgtgtTGGCGGGAGGGTGTGGGATGCTCGTGTGATTTATTACAGTGTCCATAATTCTTCGGGGGGATTCCTTTTCTCC
Hy_pMT Sema-2b nt 150-980Used in Supplemental Figure 8DTGTTCGAGCCTTATCAATAGTTGCCTCCTTGCTAACAATGTCTGTGTCATTTCTCTCTTTCCCCCTTCCCGTTAACCAAAAATGCAGTACGCCGGCACAAATGCCGAGTTCACCAAAGCGGATTCCGTCATATTCCGCTCGGACCTGTACAACCTGACGAATGGACGCAAGGAGGCCAACTTCAAGAGAACCGTGAAGTACGACTCCAAGCTGCTGGACAGTAAGTAGTGGCCATAAATCATATATTGCTAAGACAACTTTTAATGATTACATTTCTATTCATGGAAATGTGTAAAATCCAAATAATATGTAACAAATCACACATCAATCGCTTTTCTTTTCAGAGCCCAACTTCGTCGGCTCCTTTGAGATTGGTGAATTTGTGTACTTCTTCTTCCGCGAACATGCCGTTGAGTACATAAATTGCGGAAAAGCCGTCTACTCCCGAGTGGCTCGGGTGTGCAAAAACGACCGCGGTGGAAAATACATGATCAGCCAGAATTGGGCCACGTATCTGAAGGCGCGGATGAACTGCAGCATCTCCAGCGAATTTCCCTTCTACTTCAATGAGATACAATCCGTCTACAAGATGCCAACGGACGATACCAAGTTCTATGCCACGTTCACAACGAATACGAATGGTCTGATTGGATCGGCAGTGTGCAGCTACGACATCCGGGACATCAATGCGGCCTTCGACGGTGAGTATTCACTGTAGATATAGGTCGgtgtctgtgtgagtgtgtgtgtgtgtgtgtTGGCGGGAGGGTGTGGGATGCTCGTGTGATTTATTACAGTGTCCATAATTCTTCGGGGGGATTCCTTTTCTCC
Hy_pMT Sema-2b nt 1-930Used in Supplemental Figure 8DGAGCACAAGGCACAAGcacacacacatacacacacacacatgcacacatacacaTGAGCAGATGAGTTCACTCAGAGAGGGAGGCACGTGCGTACACTTTATGCGTAATTGTGTGCGTGCGTGCCCTGGCTTACTGGGCGTATGCGTGATGTTCGAGCCTTATCAATAGTTGCCTCCTTGCTAACAATGTCTGTGTCATTTCTCTCTTTCCCCCTTCCCGTTAACCAAAAATGCAGTACGCCGGCACAAATGCCGAGTTCACCAAAGCGGATTCCGTCATATTCCGCTCGGACCTGTACAACCTGACGAATGGACGCAAGGAGGCCAACTTCAAGAGAACCGTGAAGTACGACTCCAAGCTGCTGGACAGTAAGTAGTGGCCATAAATCATATATTGCTAAGACAACTTTTAATGATTACATTTCTATTCATGGAAATGTGTAAAATCCAAATAATATGTAACAAATCACACATCAATCGCTTTTCTTTTCAGAGCCCAACTTCGTCGGCTCCTTTGAGATTGGTGAATTTGTGTACTTCTTCTTCCGCGAACATGCCGTTGAGTACATAAATTGCGGAAAAGCCGTCTACTCCCGAGTGGCTCGGGTGTGCAAAAACGACCGCGGTGGAAAATACATGATCAGCCAGAATTGGGCCACGTATCTGAAGGCGCGGATGAACTGCAGCATCTCCAGCGAATTTCCCTTCTACTTCAATGAGATACAATCCGTCTACAAGATGCCAACGGACGATACCAAGTTCTATGCCACGTTCACAACGAATACGAATGGTCTGATTGGATCGGCAGTGTGCAGCTACGACATCCGGGACATCAATGCGGCCTTCGACGGTGAGTATTCACTGTAGATATAGGTCGgtgtctgtgtgagtgtgtgtgtgtgtgtgtTGGCGGGAGGGTGTGGGATGCTC
Hy_pMT Sema-2b nt 1-880Used in Supplemental Figure 8DGAGCACAAGGCACAAGcacacacacatacacacacacacatgcacacatacacaTGAGCAGATGAGTTCACTCAGAGAGGGAGGCACGTGCGTACACTTTATGCGTAATTGTGTGCGTGCGTGCCCTGGCTTACTGGGCGTATGCGTGATGTTCGAGCCTTATCAATAGTTGCCTCCTTGCTAACAATGTCTGTGTCATTTCTCTCTTTCCCCCTTCCCGTTAACCAAAAATGCAGTACGCCGGCACAAATGCCGAGTTCACCAAAGCGGATTCCGTCATATTCCGCTCGGACCTGTACAACCTGACGAATGGACGCAAGGAGGCCAACTTCAAGAGAACCGTGAAGTACGACTCCAAGCTGCTGGACAGTAAGTAGTGGCCATAAATCATATATTGCTAAGACAACTTTTAATGATTACATTTCTATTCATGGAAATGTGTAAAATCCAAATAATATGTAACAAATCACACATCAATCGCTTTTCTTTTCAGAGCCCAACTTCGTCGGCTCCTTTGAGATTGGTGAATTTGTGTACTTCTTCTTCCGCGAACATGCCGTTGAGTACATAAATTGCGGAAAAGCCGTCTACTCCCGAGTGGCTCGGGTGTGCAAAAACGACCGCGGTGGAAAATACATGATCAGCCAGAATTGGGCCACGTATCTGAAGGCGCGGATGAACTGCAGCATCTCCAGCGAATTTCCCTTCTACTTCAATGAGATACAATCCGTCTACAAGATGCCAACGGACGATACCAAGTTCTATGCCACGTTCACAACGAATACGAATGGTCTGATTGGATCGGCAGTGTGCAGCTACGACATCCGGGACATCAATGCGGCCTTCGACGGTGAGTATTCACTGTAGATATAGGTCGgtg
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The following plasmids were generated by inserting the indicated sequences between the NotI and SpeI sites of the Hy_pMT Laccase2 Sense Vector.
Hy_pMT Laccase2 1-1945 dH3 SenseUsed in Figure 5BGCTGACACGGCATTAACTTGCAGATAAAGCGCTAGCGTACTCTATAATCGGTCCTTTTCAGGACCACAAACCAGATTCAATGAGATAAAATTTTCTGTTGCCGACTATTT
Hy_pMT Laccase2 1-1945 dH3 AntisenseUsed in Figure 5BAAATAGTCGGCAACAGAAAATTTTATCTCATTGAATCTGGTTTGTGGTCCTGAAAAGGACCGATTATAGAGTACGCTAGCGCTTTATCTGCAAGTTAATGCCGTGTCAGC
Hy_pMT Laccase2 1-1945 mMALAT1_3’ mascRNAUsed in Figure 5BGATTCGTCAGTAGGGTTGTAAAGGTTTTTCTTTTCCTGAGAAAACAACCTTTTGTTTTCTCAGGTTTTGCTTTTTGGCCTTTCCCTAGCTTTAAAAAAAAAAAAGCAAAA
Hy_pMT Laccase2 1-1945 HhRzUsed in Figure 5BCCTGTCACCGGATGTGTTTTCCGGTCTGATGAGTCCGTGAGGACGAAACAGG
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The following plasmids were generated by inserting the indicated sequences between the NotI and SpeI sites of the Hy_pMT EGFP SV40 pA Sense Vector.
Hy_pMT EGFP dH3 SenseUsed in Figure 5BGCTGACACGGCATTAACTTGCAGATAAAGCGCTAGCGTACTCTATAATCGGTCCTTTTCAGGACCACAAACCAGATTCAATGAGATAAAATTTTCTGTTGCCGACTATTT
Hy_pMT EGFP dH3 AntisenseUsed in Figure 5BAAATAGTCGGCAACAGAAAATTTTATCTCATTGAATCTGGTTTGTGGTCCTGAAAAGGACCGATTATAGAGTACGCTAGCGCTTTATCTGCAAGTTAATGCCGTGTCAGC
Hy_pMT EGFP mMALAT1_3’ mascRNAUsed in Figure 5BGATTCGTCAGTAGGGTTGTAAAGGTTTTTCTTTTCCTGAGAAAACAACCTTTTGTTTTCTCAGGTTTTGCTTTTTGGCCTTTCCCTAGCTTTAAAAAAAAAAAAGCAAAA
Hy_pMT EGFP HhRzUsed in Figure 5BCCTGTCACCGGATGTGTTTTCCGGTCTGATGAGTCCGTGAGGACGAAACAGG
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The following plasmids were generated by inserting the indicated sequences between the KpnI and XmaI sites of the Hy_pMT Laccase2 MCS Exon Vector.
Hy_pMT Laccase2 MCS-Laccase2 608-707Used in Figure 6BAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAAC
Hy_pMT Laccase2 MCS-Laccase2 608-807Used in Figure 6BAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATC
Hy_pMT Laccase2 MCS-Laccase2 608-907Used in Figure 6BAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAG
Hy_pMT Laccase2 MCS-Laccase2 608-1007Used in Figure 6BAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCC
Hy_pMT Laccase2 MCS-Laccase2 608-1097Used in Figures 6B, 6CUsed in Supplemental Figure 9AAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGC
Hy_pMT Laccase2 MCS-Laccase2 708-1097Used in Figure 6CACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGC
Hy_pMT Laccase2 MCS-Laccase2 808-1097Used in Figure 6CCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGC
Hy_pMT Laccase2 MCS-Laccase2 908-1097Used in Figure 6CGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGC
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Hy_pMT Laccase2 MCS-Laccase2 1008-1097Used in Figure 6CTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGC
Hy_pMT Laccase2 MCS-ZKSCAN1 548-847Used in Figure 6DUsed in Supplemental Figure 5AGAATAGTAAAGAAACACATCATAAAACCTCCCAGGACATAAAGGTGAGCACAGACCCTGTTTGGATCAAGTCAGTTCCTGGAGCCTGAATGATGACTGCTGAATCACGGGAAGCCACGGGTCTGTCCCCACAGGCTGCACAGGAGAAGGATGGTATCGTAATAGTGAAGGTGGAAGAGGAAGATGAGGAAGACCACATGTGGGGGCAGGATTCCACCCTACAGGACACGCCTCCTCCAGACCCAGAGATATTCCGCCAACGCTTCAGGCGCTTCTGTTACCAGAACACTTTTGGGCCCCG
Hy_pMT Laccase2 MCS-ZKSCAN1 548-1047Used in Figure 6D Used in Supplemental Figure 5AGAATAGTAAAGAAACACATCATAAAACCTCCCAGGACATAAAGgtgagcacagaccctgtttggatcaagtcagttcctggagcctgaatgatgactgctgaatcacgggaagccacgggtctgtccccacagGCTGCACAGGAGAAGGATGGTATCGTAATAGTGAAGGTGGAAGAGGAAGATGAGGAAGACCACATGTGGGGGCAGGATTCCACCCTACAGGACACGCCTCCTCCAGACCCAGAGATATTCCGCCAACGCTTCAGGCGCTTCTGTTACCAGAACACTTTTGGGCCCCGAGAGGCTCTCAGTCGGCTGAAGGAACTTTGTCATCAGTGGCTGCGGCCAGAAATAAACACCAAGGAACAGATCCTGGAGCTTCTGGTGCTAGAGCAGTTTCTTTCCATCCTGCCCAAGGAGCTCCAGGTCTGGCTGCAGGAATACCGCCCCGATAGTGGAGAGGAGGCCGTGACCCTTCTAGAAGACTTGGAGCTTGATT
Hy_pMT Laccase2 MCS-ZKSCAN1 548-1417 (No Intron)Used in Figure 6DUsed in Supplemental Figure 5AGAATAGTAAAGAAACACATCATAAAACCTCCCAGGACATAAAGgtgagcacagaccctgtttggatcaagtcagttcctggagcctgaatgatgactgctgaatcacgggaagccacgggtctgtccccacagGCTGCACAGGAGAAGGATGGTATCGTAATAGTGAAGGTGGAAGAGGAAGATGAGGAAGACCACATGTGGGGGCAGGATTCCACCCTACAGGACACGCCTCCTCCAGACCCAGAGATATTCCGCCAACGCTTCAGGCGCTTCTGTTACCAGAACACTTTTGGGCCCCGAGAGGCTCTCAGTCGGCTGAAGGAACTTTGTCATCAGTGGCTGCGGCCAGAAATAAACACCAAGGAACAGATCCTGGAGCTTCTGGTGCTAGAGCAGTTTCTTTCCATCCTGCCCAAGGAGCTCCAGGTCTGGCTGCAGGAATACCGCCCCGATAGTGGAGAGGAGGCCGTGACCCTTCTAGAAGACTTGGAGCTTGATTTATCAGGACAACAGGTCCCAGGTCAAGTTCATGGACCTGAGATGCTCGCAAGGGGGATGGTGCCTCTGGATCCAGTTCAGGAGTCCTCGAGCTTTGACCTTCATCACGAGGCCACCCAGTCCCACTTCAAACATTCGTCTCGGAAACCC
Hy_pMT Laccase2 MCS-HIPK3 ExonUsed in Figure 6EUsed in Supplemental Figure 5Agtatggcctcacaagtcttggtctacccaccatatgtttatcaaactcagtcaagtgccttttgtagtgtgaagaaactcaaagtagagccaagcagttgtgtattccaggaaagaaactatccacggacctatgtgaatggtagaaactttggaaattctcatcctcccactaagggtagtgcttttcagacaaagataccatttaatagacctcgaggacacaacttttcattgcagacaagtgctgttgttttgaaaaacactgcaggtgctacaaaggtcatagcagctcaggcacagcaagctcacgtgcaggcacctcagattggggcgtggcgaaacagattgcatttcctagaaggcccccagcgatgtggattgaagcgcaagagtgaggagttggataatcatagcagcgcaatgcagattgtcgatgaattgtccatacttcctgcaatgttgcaaaccaacatgggaaatccagtgacagttgtgacagctaccacaggatcaaaacagaattgtaccactggagaaggtgactatcagttagtacagcatgaagtcttatgctccatgaaaaatacttacgaagtccttgattttcttggtcgaggcacgtttggccaggtagttaaatgctggaaaagagggacaaatgaaattgtagcaatcaaaattttgaagaatcatccttcttatgcccgtcaaggtcaaatagaagtgagcatattagcaaggctcagtactgaaaatgctgatgaatataactttgtacgagcttatgaatgctttcagcaccgtaaccatacttgtttagtctttgagatgctggaacaaaacttgtatgactttctgaaacaaaataaatttagtcccctgccactaaaagtgattcggcccattcttcaacaagtggccactgcactgaaaaaattgaaaagtcttggtttaattcatgctgatctcaagccagagaatattatgttggtggatcctgttcggcagccttacagggttaaagtaatagactttgggtcggccagtcatgtatcaaagactgtttgttcaacatatctacaatctcggtactacag
Hy_pMT Laccase2 MCS-ciRS7 ExonUsed in Figure 6FUsed in Supplemental Figure 5AGGTTTCCGATGGCACCTGTGTCAAGGTCTTCCAACAACTCCGGGTCTTCCAGCGACTTCAAGTCTTCCAATAATCTCAAGGTCTTCCAGATAATCCTGAGCTTCCAGAAAATCCACATCTTCCAGACAATCCATGTCTTCCGGACAATCCATGTCTTCCAAGAAGCTCCAAGTCTTCCAGTAAATCAAGTCTTCCAGCAAATCCAGTCTTCCAGCAATTACTGGTCTTCCACCAAATCCAGATCTTCCAGGAAAATCCACGTCTTCCAGGAAATCCATGTCTTCCAATAATTTCAAGGTCTTCCATCAAATACAGATCTTCCAGCTAATCCATGTCTTCCAGAAAAATCTGTGTCTTCCACCAAATCCAAGTCTTCCAGTAAATCTAGTTCTTCCAGAAAAATCTAGATCTTCCAGTCAATCAGTGTCTTCCAGAAAGAAATCCAGGTCTTCCAGTCAATCAGTGTCTTCCAGAAAGAAATCCAGGTCTTCCAGTCAGTCAGTGTCTTCCAGAAAAATCTACGTCTTCCACCAAATCCAGGTCTTCCAGTCAATCCACATCTTCCGGAAAAAATCCAGGTCT
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TCCAGCCAATATATGTCTTCCTGAAGATCCACGTCTTCCAGAAAATCCATGTCTTCCAGAAAATCCATGTCTTCCAGTAACCTCCCAGTCTTCCAGAAAATCCACGTCTTCCCAACAATCCAAGTCTTCCGGATAATTTGGGTCTTCCTGAAAATCTACGTCTTCCAAAAAAGCCATGTCTTCCAGAAAATCCACATCTTCCAATGGCCTCCAGGTCTTCCAGACTATCCATGTCTTCCAGAAAATCCTTGTCTTCCCTTAAATCTATAGCTTCCAAAAAATCCGGGTCTTCCAGGAAATCCGTGTCTTCCAGCAAGTCCACGTCTTCCAACAAAGCCATGTCTTCCAGACTATCCATGTCTTCCAGAAAATCCTTGTCTTCCCTCAAATCCATAGCTTCCGAAAAATCCAGGTCTTCCAGGAAATCCGTGTCTTCCAGCAAATCCACGTCTTCCAACAAAGCCATGTCTTCCATCAAATTAATGTCTTCCAGCCTACTTGTGTCTTCCAACAAAGGTACGTCTTCCAACAAAGGTACGTCTTCCAACAAAGGTATGTCTTCCAACAAAGGTACGTCTTCCAGAAAATCCACGTCTTCCAACCAAGCCATGTCTTCCAGAAAATCCACGTCTTCCAGAAAATATATGTCTTCCAACTAAGCTACGTCTTCCAACAAATCCATGTCTTCCTATATCTCCAGGTCTTCCAGCATCTCCAGGGCTTCCAGCATCTGCTCGTCTTCCAACATCTCCACGTCTTCCAGCATCTCTGTGTCTTCCAGCATCTTCATGTCTTCCAACAACTACCCAGTCTTCCATCAACTGGCTCAATATCCATGTCTTCCAACGTCTCCAGTGTGCTGATCTTCTGACATTCAGGTCTTCCAGTGTCTGCAATATCCAG
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The following plasmids were generated by inserting the indicated sequences between the EcoRI site and NotI sites of pcDNA3.1(+) (Life Technologies).
pcDNA3.1(+) Laccase2 Sense (Exon in yellow)Used in Supplemental Figure 6ACATTGAGAAATGACTGAGTTCCGGTGCTCTCAAGTCATTGATCTTTGTCGACTTTTATTTGGTCTCTGTAATAACGACTTCAAAAACATTAAATTCTGTTGCGAAGCCAGTAAGCTACAAAAAGAAAaaacaagagagaatgctatagtcgtatagtatagtttcccgactatctgatacccattacttatctagggggaatgcgaacccaaaattttatcagttttctcggatatcgatagatattggggaataaatttaaataaataaattttgggcgggtttagggcgtggcaaaaagttttttggcaaatcgctagaaatttacaagacttataaaattatgaaaaaatacaacaaaattttaaacacgtgggcgtgacagttttggacggttttagggcgttagagtaggcgaggacagggttacatcgactaggctttgatcctgatcaagaatatatatactttataccgcttccttctacatgttacctatttttcaacgaatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagattcgttgaaaagtatgtaacaggaagaataaagcatttccgaccatgtaaagtatatatattcttaataaggatcaatagccgagtcgatctcgccatgtccgtctgtcttattattttattaccgccgagacatcaggaactataaaagctagaaggatgagttttagcatacagattctagagacaaggacgcagagcaagtttgttgatccatgctgccacgctttaactttctcaaattgcccaaaactgccatgcccacatttttgaactattttcgaaattttttcataattgtattactcgtgtaaatttccatcaatttgccaaaaaactttttgtcacgcgttaacgccctaaagccgccaatttggtcacgcccacactattgaacaattatcaaattttttctcattttattccccaatatctatcgatatccccgattatgaaattattaaatttcgcgttcgcattcacactagctgagtaacgagtatctgatagttggggaaatcgactTATTTTTTATATACAATGAAAATGAATTTAATCATATGAATATCGATTATAGCTTTTTATTTAATATGAATATTTATTTGGGCTTAAGGTGTAACCTcctcgacataagactcacatggcgcaggcacattgaagacaaaaatactcaTTGTCGGGTCTCGCACCCTCCAGCAGCACCTAAAATTATGTCTTCAATTATTGCCAACATTGGAGACACAATTAGTCTGTGGCACCTCAG
pcDNA3.1(+) Laccase2 AntisenseUsed in Supplemental Figure 6ACTGAGGTGCCACAGACTAATTGTGTCTCCAATGTTGGCAATAATTGAAGACATAATTTTAGGTGCTGCTGGAGGGTGCGAGACCCGACAAtgagtatttttgtcttcaatgtgcctgcgccatgtgagtcttatgtcgaggAGGTTACACCTTAAGCCCAAATAAATATTCATATTAAATAAAAAGCTATAATCGATATTCATATGATTAAATTCATTTTCATTGTATATAAAAAATAagtcgatttccccaactatcagatactcgttactcagctagtgtgaatgcgaacgcgaaatttaataatttcataatcggggatatcgatagatattggggaataaaatgagaaaaaatttgataattgttcaatagtgtgggcgtgaccaaattggcggctttagggcgttaacgcgtgacaaaaagttttttggcaaattgatggaaatttacacgagtaatacaattatgaaaaaatttcgaaaatagttcaaaaatgtgggcatggcagttttgggcaatttgagaaagttaaagcgtggcagcatggatcaacaaacttgctctgcgtccttgtctctagaatctgtatgctaaaactcatccttctagcttttatagttcctgatgtctcggcggtaataaaataataagacagacggacatggcgagatcgactcggctattgatccttattaagaatatatatactttacatggtcggaaatgctttattcttcctgttacatacttttcaacgaatctaCAATAGTATACAATAGTATAGAAACTGCCTATTAAAAAATCGAATATTTCTAGTAAAAAATTTTGGAATTTTGAATACTTACGCTCCCAATACTGTGTAGTATTCCAACGTAAAATGATAGTAGCAAATTCTCGGAGGCTCGCCTTCGCGACAAGCGCGTGCACATTCATCAGGAGCAGATAGCGCAGGATTGCGCCGCAGCTCGGCAGTTGCACTAGTAGAGAAATCTAAGTGGCGGAATGGAGATTTTGGGTTGGGTTCCACGACGGTTCCTGTGATCTTATTAGCTGGATAACGTGCAGAGGTCGGTAGACTCCCGCGTGAGTTTAGGTAACCGCTTCCAGCGGTTAGGGCTGGCTTAGTACCAGCTAGGATTGAGGATGGAGCTCCAGACCCAAGGCTATTGCCATTTAATGGAGCGGGCCCAGAGTGGCTTCCACCTATAGACGGGTGAGTCTGCACAAGGCCGTGTGTTGCCGAAAAAAATGGTGAATCGAATGGAGTTGAACTACTAGGAGTGGCCACATGTTGCTGCCAGAAGGACCCCATTGGTGTAAGATCTGGTTGATTTTCTGCataaaataaaaaaaatatatataacaataaaaCTTAGTCTCGATTTAGCTTATTAttatacccataaatcgtacagtaaaaaggtatactagattcgttgaaaaataggtaacatgtagaaggaagcggtataaagtatatatattcttgatcaggatcaaagcctagtcgatgtaaccctgtcctcgcctactctaacgccctaaaaccgtccaaaactgtcacgcccacgtgtttaaaattttgttgtattttttcataattttataagtcttgtaaatttctagcgatttgccaaaaaactttttgccacgccctaaacccgcccaaaatttatttatttaaatttattccccaatatctatcgatatccgagaaaactgataaaattttgggttcgcattccccctagataagtaatgggtatcagatagtcgggaaactatactatacgactatagcattctctcttgtttTTTCTTTTTGTAGCTTACTGGCTTCGCAACAGAATTTAATGTTTTTGAAGTCGTTATTACAGAGACCAAATAAAAGTCGACAAAGATCAATGACTTGAGAGCACCGGAACTCAGTCATTTCTCAATG
pcDNA3.1(+) Laccase2 MCS Exon Vector (Exon in yellow)Used in Figures 7C, 7DUsed in Supplemental Figure 6BCATTGAGAAATGACTGAGTTCCGGTGCTCTCAAGTCATTGATCTTTGTCGACTTTTATTTGGTCTCTGTAATAACGACTTCAAAAACATTAAATTCTGTTGCGAAGCCAGTAAGCTACAAAAAGAAAaaacaagagagaatgctatagtcgtatagtatagtttcccgactatctgatacccattacttatctagggggaatgcgaacccaaaattttatcagttttctcggatatcgatagatattggggaataaatttaaataaataaattttgggcgggtttagggcgtggcaaaaagttttttggcaaatcgctagaaatttacaagacttataaaattatgaaaaaatacaacaaaattttaaacacgtgggcgtgacagttttggacggttttagggcgttagagtaggcgaggacagggttacatcgactaggctttgatcctgatcaagaatatatatactttataccgcttccttctacatgttacctatttttcaacgaatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAATTAATTAAACCGGTCCTGCAGGTGATCAGGCGCGCCGGTAACCGGCCGGCCCCGCGGAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagattcgttgaaaagtatgtaacaggaagaataaagcatttccgaccatgtaaagtatatatattcttaataaggatcaatagccgagtcgatctcgccatgtccgtctgtcttattattttattaccgccgagacatcaggaactataaaagctagaaggatgagttttagcatacagattctagagacaaggacgcagagcaagtttgttgatccatgctgccacgctttaactttctcaaattgcccaaaactgccatgcccacatttttgaactattttcgaaattttttcataattgtattactcgtgt
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aaatttccatcaatttgccaaaaaactttttgtcacgcgttaacgccctaaagccgccaatttggtcacgcccacactattgaacaattatcaaattttttctcattttattccccaatatctatcgatatccccgattatgaaattattaaatttcgcgttcgcattcacactagctgagtaacgagtatctgatagttggggaaatcgactTATTTTTTATATACAATGAAAATGAATTTAATCATATGAATATCGATTATAGCTTTTTATTTAATATGAATATTTATTTGGGCTTAAGGTGTAACCTcctcgacataagactcacatggcgcaggcacattgaagacaaaaatactcaTTGTCGGGTCTCGCACCCTCCAGCAGCACCTAAAATTATGTCTTCAATTATTGCCAACATTGGAGACACAATTAGTCTGTGGCACCTCAG
pcDNA3.1(+) ZKSCAN1 MCS Vector (Exon in yellow)Used in Figures 7C, 7D Used in Supplemental Figures 6B, 6C, 6FAGTGACAGTGGAGATTGTACAGTTTTTTCCTCGATTTGTCAGGAtttttttttttttgacggagtttaacttcttgtctcccaggtaggaagtgcagtggcgtaatctcggctcactacaacctccacctcctgggttcaagcgtttctcctgcctcagctttccgagtagctgggattacaggcgcctgccaccatgccctgctgacttttgtatttttagtagagacggggtttcaccatgttggccaggctggtcttgaactcctgaccgcaggcgattggcctgcctcggcctcccaaagtgctgagattacaggcgtgagccaccacccccggccTCAGGAGCGTTCTGATAGTGCCTCGATGTGCTGCCTCCTATAAAGTGTTAGCAGCACAGATCACTTTTTGTAAAGGTACGTACTAATGACTTTTTTTTTATACTTCAGGATATCATCGATACCGGTTTAATTAACACGTGGGTAACCGTTAACCCGCGGAGGTAAGAAGCAAGGTTTCATTTAGGGGAAGGGAAATGATTCAGGACGAGAGTCTTTGTGCTGCTGAGTGCCTGTGATGAAGAAGCATGTTAGTcctgggcaacgtagcgagaccccatctctacaaaaaatagaaaaattagccaggtatagtggcgcacacctgtgattccagctacgcaggaggctgaggtgggaggattgcttgagcccaggaggttgaggctgcagtgagctgtaatcatgccactactccaacctgggcaacacagcaaggaccctgtctcaaaaGCTACTTACAGAAAAGAATTAggctcggcacggtagctcacacctgtaatcccagcactttgggaggctgaggcgggcagatcacttgaggtcaggagtttgagaccagcctggccaacatggtgaaaccttgtctctactaaaaatatgaaaattagccaggcatggtggcacattcctgtaatcccagctactcgggaggctgaggcaggagaatcacttgaacccaggaggtggaggttgcagtaagccgagatcgtaccactgtgctctagccttggtgacagagcgagactgtcttaaaaaaaaaaaaaaaaaaaaaagaattaattaaaaatttaaaaaaaaatgaaaaaaaGCTGCATGCTTGTTTTTTGTTTTTAGTTATTCTACATTGTTGTCATTATTACCAAATATTGGGGAAAATACAACTTACAGACCAATCTCAGGAGTTAAATGTTACTACGAAGGCAAATGAACTATGCGTAATGAACCTGGTAGGCATTA
pcDNA3.1(+) ZKSCAN1 MCS-GFP 5’ Half Only (Exon in yellow)Used in Supplemental Figure 7BAGTGACAGTGGAGATTGTACAGTTTTTTCCTCGATTTGTCAGGAtttttttttttttgacggagtttaacttcttgtctcccaggtaggaagtgcagtggcgtaatctcggctcactacaacctccacctcctgggttcaagcgtttctcctgcctcagctttccgagtagctgggattacaggcgcctgccaccatgccctgctgacttttgtatttttagtagagacggggtttcaccatgttggccaggctggtcttgaactcctgaccgcaggcgattggcctgcctcggcctcccaaagtgctgagattacaggcgtgagccaccacccccggccTCAGGAGCGTTCTGATAGTGCCTCGATGTGCTGCCTCCTATAAAGTGTTAGCAGCACAGATCACTTTTTGTAAAGGTACGTACTAATGACTTTTTTTTTATACTTCAGGATATCATCGATACCGGTTTAATTAACACGTGGGTAACCGTTCCGAGTCGTTAACCACCatggccagcaagggcgCCgagctgttcaccggcAtCgtgcccatcctgAtCgagctgAaTggcgatgtgaatggccacaagttcagcgtgagcggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcctgtgccctggcccaccctggtgaccaccctgagctacggcgtgcagtgcttctcacgctaccccgatcacatgaagcagcacgacttcttcaagagcgccatgcctgagggctacatccaggagcgcaccatcttcttcgaggatgacggcaactacaagtcgcgcgccgaggtgaagttcgagggcgataccctggtgaatcgcatcgagctgaccggcaccgatttcaaggaggatggcaacatcctgggcaataagatggagtacaactacaacgcccacaatgtgtacatcatgaccgacaaggccaagaatggcatcaaggtgaacttcaagatccgccacaCCGCGGAGGTAAGAAGCAAGGTTTCATTTAGGGGAAGGGAAATGATTCAGGACGAGAGTCTTTGTGCTGCTGAGTGCCTGTGATGAAGAAGCATGTTAGTcctgggcaacgtagcgagaccccatctctacaaaaaatagaaaaattagccaggtatagtggcgcacacctgtgattccagctacgcaggaggctgaggtgggaggattgcttgagcccaggaggttgaggctgcagtgagctgtaatcatgccactactccaacctgggcaacacagcaaggaccctgtctcaaaaGCTACTTACAGAAAAGAATTAggctcggcacggtagctcacacctgtaatcccagcactttgggaggctgaggcgggcagatcacttgaggtcaggagtttgagaccagcctggccaacatggtgaaaccttgtctctactaaaaatatgaaaattagccaggcatggtggcacattcctgtaatcccagctactcgggaggctgaggcaggagaatcacttgaacccaggaggtggaggttgcagtaagccgagatcgtaccactgtgctctagccttggtgacagagcgagactgtcttaaaaaaaaaaaaaaaaaaaaaagaattaattaaaaatttaaaaaaaaatgaaaaaaaGCTGCATGCTTGTTTTTTGTTTTTAGTTATTCTACATTGTTGTCATTATTACCAAATATTGGGGAAAATACAACTTACAGACCAATCTCAGGAGTTAAATGTTACTACGAAGGCAAATGAACTATGCGTAATGAACCTGGTAGGCATTA
pcDNA3.1(+) ZKSCAN1 MCS-WT Split GFP (Exon in yellow)Used in Supplemental Figures 7B, 7CAGTGACAGTGGAGATTGTACAGTTTTTTCCTCGATTTGTCAGGAtttttttttttttgacggagtttaacttcttgtctcccaggtaggaagtgcagtggcgtaatctcggctcactacaacctccacctcctgggttcaagcgtttctcctgcctcagctttccgagtagctgggattacaggcgcctgccaccatgccctgctgacttttgtatttttagtagagacggggtttcaccatgttggccaggctggtcttgaactcctgaccgcaggcgattggcctgcctcggcctcccaaagtgctgagattacaggcgtgagccaccacccccggccTCAGGAGCGTTCTGATAGTGCCTCGATGTGCTGCCTCCTATAAAGTGTTAGCAGCACAGATCACTTTTTGTAAAGGTACGTACTAATGACTTTTTTTTTATACTTCAGGATggcagcgtgcagctggccgaccactaccagcagaatacccccatcggcgatggccctgtgctgctgcccgataaccactacctgtccacccagagcgccctgtccaaggaccccaacgagaagcgcgatcacatgatCTACTTCGGCttcgtgaccgccgccggcatcacccacggcatggacgagctgtacaagtgaATCGATACCGGTTTAATTAACACGTGGGTAACCGTTCCGAGTCGTTAACCACCatggccagcaagggcgCCgagctgttcaccggcAtCgtgcccatcctgAtCgagctgAaTggcgatgtgaatggccacaagttcagcgtgagcggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcctgtgccctggcccaccctggtgaccaccctgagctacggcgtgcagtgcttctcacgctaccccgatcacatgaagcagcacgacttcttcaagagcgccatgcctgagggctacatccaggagcgcaccatcttcttcgaggatgacggcaactacaagtcgcgcgccgaggtgaagttcgagggcgataccctggtgaatcgcatcgagctgaccggcaccgatttcaaggaggatggcaacatcctgggcaataagatggagtacaactacaacgcccacaatgtgtacatcatgaccgacaaggccaagaatggcatcaaggtgaacttcaagatccgccacaACATCGAGGTAAGAAGCAAGGTTTCATTTAGGGGAAGGGAAATGATTCAGGACGAGAGTCTTTGTGCTGCTGAGTGCCTGTGATGAAGAAGCATGTTAGTcctgggcaacgtagcgagaccccatctctacaaaaaatagaaaaattagccaggtatagtggcgcacacctgtgattccagctacgcaggaggctgaggtgggaggattgcttgagcccaggaggttgaggctgcagtgagctgtaatcatgccactactccaacctgggcaacacagcaaggaccctgtctcaaaaGCTACTTACAGAAAAGAATTAggctcggcacggtagctcacacctgtaatcccagcactttgggaggctgaggcgggcagatcacttgaggtcaggagtttgagaccagcctggccaacatggtgaaaccttgtctctactaaaaatatgaaaattagccaggcatggtggcacattcctgtaatcccagctactcgggaggctgaggcaggagaatcacttgaacccaggaggtg
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gaggttgcagtaagccgagatcgtaccactgtgctctagccttggtgacagagcgagactgtcttaaaaaaaaaaaaaaaaaaaaaagaattaattaaaaatttaaaaaaaaatgaaaaaaaGCTGCATGCTTGTTTTTTGTTTTTAGTTATTCTACATTGTTGTCATTATTACCAAATATTGGGGAAAATACAACTTACAGACCAATCTCAGGAGTTAAATGTTACTACGAAGGCAAATGAACTATGCGTAATGAACCTGGTAGGCATTA
pcDNA3.1(+) ZKSCAN1 MCS-WT Split GFP + Sense IRES (Exon in yellow)Used in Supplemental Figures 7B, 7CAGTGACAGTGGAGATTGTACAGTTTTTTCCTCGATTTGTCAGGAtttttttttttttgacggagtttaacttcttgtctcccaggtaggaagtgcagtggcgtaatctcggctcactacaacctccacctcctgggttcaagcgtttctcctgcctcagctttccgagtagctgggattacaggcgcctgccaccatgccctgctgacttttgtatttttagtagagacggggtttcaccatgttggccaggctggtcttgaactcctgaccgcaggcgattggcctgcctcggcctcccaaagtgctgagattacaggcgtgagccaccacccccggccTCAGGAGCGTTCTGATAGTGCCTCGATGTGCTGCCTCCTATAAAGTGTTAGCAGCACAGATCACTTTTTGTAAAGGTACGTACTAATGACTTTTTTTTTATACTTCAGGATggcagcgtgcagctggccgaccactaccagcagaatacccccatcggcgatggccctgtgctgctgcccgataaccactacctgtccacccagagcgccctgtccaaggaccccaacgagaagcgcgatcacatgatCTACTTCGGCttcgtgaccgccgccggcatcacccacggcatggacgagctgtacaagtgaATCGATACCGGTTTAATTAACACGTGGGTAACCGTTCCGAGTCGTTAACGCCCCTCTCCCTCCCCCCCCCCTAACGTTACTGGCCGAAGCCGCTTGGAATAAGGCCGGTGTGCGTTTGTCTATATGTTATTTTCCACCATATTGCCGTCTTTTGGCAATGTGAGGGCCCGGAAACCTGGCCCTGTCTTCTTGACGAGCATTCCTAGGGGTCTTTCCCCTCTCGCCAAAGGAATGCAAGGTCTGTTGAATGTCGTGAAGGAAGCAGTTCCTCTGGAAGCTTCTTGAAGACAAACAACGTCTGTAGCGACCCTTTGCAGGCAGCGGAACCCCCCACCTGGCGACAGGTGCCTCTGCGGCCAAAAGCCACGTGTATAAGATACACCTGCAAAGGCGGCACAACCCCAGTGCCACGTTGTGAGTTGGATAGTTGTGGAAAGAGTCAAATGGCTCTCCTCAAGCGTATTCAACAAGGGGCTGAAGGATGCCCAGAAGGTACCCCATTGTATGGGATCTGATCTGGGGCCTCGGTGCACATGCTTTACATGTGTTTAGTCGAGGTTAAAAAAACGTCTAGGCCCCCCGAACCACGGGGACGTGGTTTTCCTTTGAAAAACACGATGATAATATGGCGTTAACCACCatggccagcaagggcgCCgagctgttcaccggcAtCgtgcccatcctgAtCgagctgAaTggcgatgtgaatggccacaagttcagcgtgagcggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcctgtgccctggcccaccctggtgaccaccctgagctacggcgtgcagtgcttctcacgctaccccgatcacatgaagcagcacgacttcttcaagagcgccatgcctgagggctacatccaggagcgcaccatcttcttcgaggatgacggcaactacaagtcgcgcgccgaggtgaagttcgagggcgataccctggtgaatcgcatcgagctgaccggcaccgatttcaaggaggatggcaacatcctgggcaataagatggagtacaactacaacgcccacaatgtgtacatcatgaccgacaaggccaagaatggcatcaaggtgaacttcaagatccgccacaACATCGAGGTAAGAAGCAAGGTTTCATTTAGGGGAAGGGAAATGATTCAGGACGAGAGTCTTTGTGCTGCTGAGTGCCTGTGATGAAGAAGCATGTTAGTcctgggcaacgtagcgagaccccatctctacaaaaaatagaaaaattagccaggtatagtggcgcacacctgtgattccagctacgcaggaggctgaggtgggaggattgcttgagcccaggaggttgaggctgcagtgagctgtaatcatgccactactccaacctgggcaacacagcaaggaccctgtctcaaaaGCTACTTACAGAAAAGAATTAggctcggcacggtagctcacacctgtaatcccagcactttgggaggctgaggcgggcagatcacttgaggtcaggagtttgagaccagcctggccaacatggtgaaaccttgtctctactaaaaatatgaaaattagccaggcatggtggcacattcctgtaatcccagctactcgggaggctgaggcaggagaatcacttgaacccaggaggtggaggttgcagtaagccgagatcgtaccactgtgctctagccttggtgacagagcgagactgtcttaaaaaaaaaaaaaaaaaaaaaagaattaattaaaaatttaaaaaaaaatgaaaaaaaGCTGCATGCTTGTTTTTTGTTTTTAGTTATTCTACATTGTTGTCATTATTACCAAATATTGGGGAAAATACAACTTACAGACCAATCTCAGGAGTTAAATGTTACTACGAAGGCAAATGAACTATGCGTAATGAACCTGGTAGGCATTA
pcDNA3.1(+) ZKSCAN1 MCS-WT Split GFP + Antisense IRES (Exon in yellow)Used in Supplemental Figures 7B, 7CAGTGACAGTGGAGATTGTACAGTTTTTTCCTCGATTTGTCAGGAtttttttttttttgacggagtttaacttcttgtctcccaggtaggaagtgcagtggcgtaatctcggctcactacaacctccacctcctgggttcaagcgtttctcctgcctcagctttccgagtagctgggattacaggcgcctgccaccatgccctgctgacttttgtatttttagtagagacggggtttcaccatgttggccaggctggtcttgaactcctgaccgcaggcgattggcctgcctcggcctcccaaagtgctgagattacaggcgtgagccaccacccccggccTCAGGAGCGTTCTGATAGTGCCTCGATGTGCTGCCTCCTATAAAGTGTTAGCAGCACAGATCACTTTTTGTAAAGGTACGTACTAATGACTTTTTTTTTATACTTCAGGATggcagcgtgcagctggccgaccactaccagcagaatacccccatcggcgatggccctgtgctgctgcccgataaccactacctgtccacccagagcgccctgtccaaggaccccaacgagaagcgcgatcacatgatCTACTTCGGCttcgtgaccgccgccggcatcacccacggcatggacgagctgtacaagtgaATCGATACCGGTTTAATTAACACGTGGGTAACCGTTCCGAGTCGTTAACGCCATATTATCATCGTGTTTTTCAAAGGAAAACCACGTCCCCGTGGTTCGGGGGGCCTAGACGTTTTTTTAACCTCGACTAAACACATGTAAAGCATGTGCACCGAGGCCCCAGATCAGATCCCATACAATGGGGTACCTTCTGGGCATCCTTCAGCCCCTTGTTGAATACGCTTGAGGAGAGCCATTTGACTCTTTCCACAACTATCCAACTCACAACGTGGCACTGGGGTTGTGCCGCCTTTGCAGGTGTATCTTATACACGTGGCTTTTGGCCGCAGAGGCACCTGTCGCCAGGTGGGGGGTTCCGCTGCCTGCAAAGGGTCGCTACAGACGTTGTTTGTCTTCAAGAAGCTTCCAGAGGAACTGCTTCCTTCACGACATTCAACAGACCTTGCATTCCTTTGGCGAGAGGGGAAAGACCCCTAGGAATGCTCGTCAAGAAGACAGGGCCAGGTTTCCGGGCCCTCACATTGCCAAAAGACGGCAATATGGTGGAAAATAACATATAGACAAACGCACACCGGCCTTATTCCAAGCGGCTTCGGCCAGTAACGTTAGGGGGGGGGGAGGGAGAGGGGCGTTAACCACCatggccagcaagggcgCCgagctgttcaccggcAtCgtgcccatcctgAtCgagctgAaTggcgatgtgaatggccacaagttcagcgtgagcggcgagggcgagggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgcctgtgccctggcccaccctggtgaccaccctgagctacggcgtgcagtgcttctcacgctaccccgatcacatgaagcagcacgacttcttcaagagcgccatgcctgagggctacatccaggagcgcaccatcttcttcgaggatgacggcaactacaagtcgcgcgccgaggtgaagttcgagggcgataccctggtgaatcgcatcgagctgaccggcaccgatttcaaggaggatggcaacatcctgggcaataagatggagtacaactacaacgcccacaatgtgtacatcatgaccgacaaggccaagaatggcatcaaggtgaacttcaagatccgccacaACATCGAGGTAAGAAGCAAGGTTTCATTTAGGGGAAGGGAAATGATTCAGGACGAGAGTCTTTGTGCTGCTGAGTGCCTGTGATGAAGAAGCATGTTAGTcctgggcaacgtagcgagaccccatctctacaaaaaatagaaaaattagccaggtatagtggcgcacacctgtgattccagctacgcaggaggctgaggtgggaggattgcttgagcccaggaggttgaggctgcagtgagctgtaatcatgccactactccaacctgggcaacacagcaaggaccctgtctcaaaaGCTACTTACAGAAAAGAATTAggctcggcacggtagctcacacctgtaatcccagcactttgggaggctgaggcgggcagatcacttgaggtcaggagtttgagaccagcctggccaacatggtgaaaccttgtctctactaaaaatatgaaaattagccaggcatggtggcacattcctgtaatcccagctactcgggaggctgaggcaggagaatcacttgaacccaggaggtggaggttgcagtaagccgagatcgtaccactgtgctctagccttggtgacagagcgagactgtcttaaaaaaaaaaaaaaaaaaaaaagaattaattaaaaatttaaaaaaaaatgaaaaaaaGCTGCATGCTTGTTTTTTGTTTTTAGTTATTCTACATTGTTGTCATTATTACCAAATATTGGGGAAAATACAACTTACAGACCAATCTCAGGAGTTAAATGTTACTACGAAGGCAAATGAACTATGCGTAATGAACCTGGTAGGCATTA
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The following plasmids were generated by inserting the indicated sequences into the PacI and SacII sites of pcDNA3.1(+) Laccase2 MCS Exon Vector.
pcDNA3.1(+) Laccase2 MCS Exon Vector + ZKSCAN1 548-847Used in Figure 7CGAATAGTAAAGAAACACATCATAAAACCTCCCAGGACATAAAGGTGAGCACAGACCCTGTTTGGATCAAGTCAGTTCCTGGAGCCTGAATGATGACTGCTGAATCACGGGAAGCCACGGGTCTGTCCCCACAGGCTGCACAGGAGAAGGATGGTATCGTAATAGTGAAGGTGGAAGAGGAAGATGAGGAAGACCACATGTGGGGGCAGGATTCCACCCTACAGGACACGCCTCCTCCAGACCCAGAGATATTCCGCCAACGCTTCAGGCGCTTCTGTTACCAGAACACTTTTGGGCCCCG
pcDNA3.1(+) Laccase2 MCS Exon Vector + ZKSCAN1 548-1047Used in Figure 7CGAATAGTAAAGAAACACATCATAAAACCTCCCAGGACATAAAGgtgagcacagaccctgtttggatcaagtcagttcctggagcctgaatgatgactgctgaatcacgggaagccacgggtctgtccccacagGCTGCACAGGAGAAGGATGGTATCGTAATAGTGAAGGTGGAAGAGGAAGATGAGGAAGACCACATGTGGGGGCAGGATTCCACCCTACAGGACACGCCTCCTCCAGACCCAGAGATATTCCGCCAACGCTTCAGGCGCTTCTGTTACCAGAACACTTTTGGGCCCCGAGAGGCTCTCAGTCGGCTGAAGGAACTTTGTCATCAGTGGCTGCGGCCAGAAATAAACACCAAGGAACAGATCCTGGAGCTTCTGGTGCTAGAGCAGTTTCTTTCCATCCTGCCCAAGGAGCTCCAGGTCTGGCTGCAGGAATACCGCCCCGATAGTGGAGAGGAGGCCGTGACCCTTCTAGAAGACTTGGAGCTTGATT
pcDNA3.1(+) Laccase2 MCS Exon Vector + ZKSCAN1 548-1417 (No Intron)Used in Figure 7CGAATAGTAAAGAAACACATCATAAAACCTCCCAGGACATAAAGgtgagcacagaccctgtttggatcaagtcagttcctggagcctgaatgatgactgctgaatcacgggaagccacgggtctgtccccacagGCTGCACAGGAGAAGGATGGTATCGTAATAGTGAAGGTGGAAGAGGAAGATGAGGAAGACCACATGTGGGGGCAGGATTCCACCCTACAGGACACGCCTCCTCCAGACCCAGAGATATTCCGCCAACGCTTCAGGCGCTTCTGTTACCAGAACACTTTTGGGCCCCGAGAGGCTCTCAGTCGGCTGAAGGAACTTTGTCATCAGTGGCTGCGGCCAGAAATAAACACCAAGGAACAGATCCTGGAGCTTCTGGTGCTAGAGCAGTTTCTTTCCATCCTGCCCAAGGAGCTCCAGGTCTGGCTGCAGGAATACCGCCCCGATAGTGGAGAGGAGGCCGTGACCCTTCTAGAAGACTTGGAGCTTGATTTATCAGGACAACAGGTCCCAGGTCAAGTTCATGGACCTGAGATGCTCGCAAGGGGGATGGTGCCTCTGGATCCAGTTCAGGAGTCCTCGAGCTTTGACCTTCATCACGAGGCCACCCAGTCCCACTTCAAACATTCGTCTCGGAAACCC
pcDNA3.1(+) Laccase2 MCS Exon Vector + ciRS7 ExonUsed in Figure 7DGGTTTCCGATGGCACCTGTGTCAAGGTCTTCCAACAACTCCGGGTCTTCCAGCGACTTCAAGTCTTCCAATAATCTCAAGGTCTTCCAGATAATCCTGAGCTTCCAGAAAATCCACATCTTCCAGACAATCCATGTCTTCCGGACAATCCATGTCTTCCAAGAAGCTCCAAGTCTTCCAGTAAATCAAGTCTTCCAGCAAATCCAGTCTTCCAGCAATTACTGGTCTTCCACCAAATCCAGATCTTCCAGGAAAATCCACGTCTTCCAGGAAATCCATGTCTTCCAATAATTTCAAGGTCTTCCATCAAATACAGATCTTCCAGCTAATCCATGTCTTCCAGAAAAATCTGTGTCTTCCACCAAATCCAAGTCTTCCAGTAAATCTAGTTCTTCCAGAAAAATCTAGATCTTCCAGTCAATCAGTGTCTTCCAGAAAGAAATCCAGGTCTTCCAGTCAATCAGTGTCTTCCAGAAAGAAATCCAGGTCTTCCAGTCAGTCAGTGTCTTCCAGAAAAATCTACGTCTTCCACCAAATCCAGGTCTTCCAGTCAATCCACATCTTCCGGAAAAAATCCAGGTCTTCCAGCCAATATATGTCTTCCTGAAGATCCACGTCTTCCAGAAAATCCATGTCTTCCAGAAAATCCATGTCTTCCAGTAACCTCCCAGTCTTCCAGAAAATCCACGTCTTCCCAACAATCCAAGTCTTCCGGATAATTTGGGTCTTCCTGAAAATCTACGTCTTCCAAAAAAGCCATGTCTTCCAGAAAATCCACATCTTCCAATGGCCTCCAGGTCTTCCAGACTATCCATGTCTTCCAGAAAATCCTTGTCTTCCCTTAAATCTATAGCTTCCAAAAAATCCGGGTCTTCCAGGAAATCCGTGTCTTCCAGCAAGTCCACGTCTTCCAACAAAGCCATGTCTTCCAGACTATCCATGTCTTCCAGAAAATCCTTGTCTTCCCTCAAATCCATAGCTTCCGAAAAATCCAGGTCTTCCAGGAAATCCGTGTCTTCCAGCAAATCCACGTCTTCCAACAAAGCCATGTCTTCCATCAAATTAATGTCTTCCAGCCTACTTGTGTCTTCCAACAAAGGTACGTCTTCCAACAAAGGTACGTCTTCCAACAAAGGTATGTCTTCCAACAAAGGTACGTCTTCCAGAAAATCCACGTCTTCCAACCAAGCCATGTCTTCCAGAAAATCCACGTCTTCCAGAAAATATATGTCTTCCAACTAAGCTACGTCTTCCAACAAATCCATGTCTTCCTATATCTCCAGGTCTTCCAGCATCTCCAGGGCTTCCAGCATCTGCTCGTCTTCCAACATCTCCACGTCTTCCAGCATCTCTGTGTCTTCCAGCATCTTCATGTCTTCCAACAACTACCCAGTCTTCCATCAACTGGCTCAATATCCATGTCTTCCAACGTCTCCAGTGTGCTGATCTTCTGACATTCAGGTCTTCCAGTGTCTGCAATATCCAG
pcDNA3.1(+) Laccase2 MCS Exon Vector + HIPK3 ExonUsed in Supplemental Figure 6Bgtatggcctcacaagtcttggtctacccaccatatgtttatcaaactcagtcaagtgccttttgtagtgtgaagaaactcaaagtagagccaagcagttgtgtattccaggaaagaaactatccacggacctatgtgaatggtagaaactttggaaattctcatcctcccactaagggtagtgcttttcagacaaagataccatttaatagacctcgaggacacaacttttcattgcagacaagtgctgttgttttgaaaaacactgcaggtgctacaaaggtcatagcagctcaggcacagcaagctcacgtgcaggcacctcagattggggcgtggcgaaacagattgcatttcctagaaggcccccagcgatgtggattgaagcgcaagagtgaggagttggataatcatagcagcgcaatgcagattgtcgatgaattgtccatacttcctgcaatgttgcaaaccaacatgggaaatccagtgacagttgtgacagctaccacaggatcaaaacagaattgtaccactggagaaggtgactatcagttagtacagcatgaagtcttatgctccatgaaaaatacttacgaagtccttgattttcttggtcgaggcacgtttggccaggtagttaaatgctggaaaagagggacaaatgaaattgtagcaatcaaaattttgaagaatcatccttcttatgcccgtcaaggtcaaatagaagtgagcatattagcaaggctcagtactgaaaatgctgatgaatataactttgtacgagcttatgaatgctttcagcaccgtaaccatacttgtttagtctttgagatgctggaacaaaacttgtatgactttctgaaacaaaataaatttagtcccctgccactaaaagtgattcggcccattcttcaacaagtggccactgcactgaaaaaattgaaaagtcttggtttaattcatgctgatctcaagccagagaatattatgttggtggatcctgttcggcagccttacagggttaaagtaatagactttgggtcggccagtcatgtatcaaagactgtttgttcaacatatctacaatctcggtactacag
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The following plasmids were generated by inserting the indicated sequences into the EcoRV and SacII sites of pcDNA3.1(+) ZKSCAN1 MCS Vector.
pcDNA3.1(+) ZKSCAN1 MCS Vector + ZKSCAN1 548-647Used in Supplemental Figure 6CGAATAGTAAAGAAACACATCATAAAACCTCCCAGGACATAAAGGTGAGCACAGACCCTGTTTGGATCAAGTCAGTTCCTGGAGCCTGAATGATGACTGCT
pcDNA3.1(+) ZKSCAN1 MCS Vector + ZKSCAN1 548-747Used in Supplemental Figure 6CGAATAGTAAAGAAACACATCATAAAACCTCCCAGGACATAAAGGTGAGCACAGACCCTGTTTGGATCAAGTCAGTTCCTGGAGCCTGAATGATGACTGCTGAATCACGGGAAGCCACGGGTCTGTCCCCACAGGCTGCACAGGAGAAGGATGGTATCGTAATAGTGAAGGTGGAAGAGGAAGATGAGGAAGACCACATGT
pcDNA3.1(+) ZKSCAN1 MCS Vector + ZKSCAN1 548-847Used in Figure 7CUsed in Supplemental Figures 6C, 6D, 6E, 6FGAATAGTAAAGAAACACATCATAAAACCTCCCAGGACATAAAGGTGAGCACAGACCCTGTTTGGATCAAGTCAGTTCCTGGAGCCTGAATGATGACTGCTGAATCACGGGAAGCCACGGGTCTGTCCCCACAGGCTGCACAGGAGAAGGATGGTATCGTAATAGTGAAGGTGGAAGAGGAAGATGAGGAAGACCACATGTGGGGGCAGGATTCCACCCTACAGGACACGCCTCCTCCAGACCCAGAGATATTCCGCCAACGCTTCAGGCGCTTCTGTTACCAGAACACTTTTGGGCCCCG
pcDNA3.1(+) ZKSCAN1 MCS Vector + ZKSCAN1 548-1047Used in Figure 7CUsed in Supplemental Figures 6C, 6D, 6E, 6FGAATAGTAAAGAAACACATCATAAAACCTCCCAGGACATAAAGGTGAGCACAGACCCTGTTTGGATCAAGTCAGTTCCTGGAGCCTGAATGATGACTGCTGAATCACGGGAAGCCACGGGTCTGTCCCCACAGGCTGCACAGGAGAAGGATGGTATCGTAATAGTGAAGGTGGAAGAGGAAGATGAGGAAGACCACATGTGGGGGCAGGATTCCACCCTACAGGACACGCCTCCTCCAGACCCAGAGATATTCCGCCAACGCTTCAGGCGCTTCTGTTACCAGAACACTTTTGGGCCCCGAGAGGCTCTCAGTCGGCTGAAGGAACTTTGTCATCAGTGGCTGCGGCCAGAAATAAACACCAAGGAACAGATCCTGGAGCTTCTGGTGCTAGAGCAGTTTCTTTCCATCCTGCCCAAGGAGCTCCAGGTCTGGCTGCAGGAATACCGCCCCGATAGTGGAGAGGAGGCCGTGACCCTTCTAGAAGACTTGGAGCTTGATT
pcDNA3.1(+) ZKSCAN1 MCS Vector + ZKSCAN1 548-1417 (No Intron)Used in Figure 7CUsed in Supplemental Figures 6C, 6D, 6E, 6FGAATAGTAAAGAAACACATCATAAAACCTCCCAGGACATAAAGgtgagcacagaccctgtttggatcaagtcagttcctggagcctgaatgatgactgctgaatcacgggaagccacgggtctgtccccacagGCTGCACAGGAGAAGGATGGTATCGTAATAGTGAAGGTGGAAGAGGAAGATGAGGAAGACCACATGTGGGGGCAGGATTCCACCCTACAGGACACGCCTCCTCCAGACCCAGAGATATTCCGCCAACGCTTCAGGCGCTTCTGTTACCAGAACACTTTTGGGCCCCGAGAGGCTCTCAGTCGGCTGAAGGAACTTTGTCATCAGTGGCTGCGGCCAGAAATAAACACCAAGGAACAGATCCTGGAGCTTCTGGTGCTAGAGCAGTTTCTTTCCATCCTGCCCAAGGAGCTCCAGGTCTGGCTGCAGGAATACCGCCCCGATAGTGGAGAGGAGGCCGTGACCCTTCTAGAAGACTTGGAGCTTGATTTATCAGGACAACAGGTCCCAGGTCAAGTTCATGGACCTGAGATGCTCGCAAGGGGGATGGTGCCTCTGGATCCAGTTCAGGAGTCCTCGAGCTTTGACCTTCATCACGAGGCCACCCAGTCCCACTTCAAACATTCGTCTCGGAAACCC
pcDNA3.1(+) ZKSCAN1 MCS Vector + ciRS7 ExonUsed in Figure 7DGGTTTCCGATGGCACCTGTGTCAAGGTCTTCCAACAACTCCGGGTCTTCCAGCGACTTCAAGTCTTCCAATAATCTCAAGGTCTTCCAGATAATCCTGAGCTTCCAGAAAATCCACATCTTCCAGACAATCCATGTCTTCCGGACAATCCATGTCTTCCAAGAAGCTCCAAGTCTTCCAGTAAATCAAGTCTTCCAGCAAATCCAGTCTTCCAGCAATTACTGGTCTTCCACCAAATCCAGATCTTCCAGGAAAATCCACGTCTTCCAGGAAATCCATGTCTTCCAATAATTTCAAGGTCTTCCATCAAATACAGATCTTCCAGCTAATCCATGTCTTCCAGAAAAATCTGTGTCTTCCACCAAATCCAAGTCTTCCAGTAAATCTAGTTCTTCCAGAAAAATCTAGATCTTCCAGTCAATCAGTGTCTTCCAGAAAGAAATCCAGGTCTTCCAGTCAATCAGTGTCTTCCAGAAAGAAATCCAGGTCTTCCAGTCAGTCAGTGTCTTCCAGAAAAATCTACGTCTTCCACCAAATCCAGGTCTTCCAGTCAATCCACATCTTCCGGAAAAAATCCAGGTCTTCCAGCCAATATATGTCTTCCTGAAGATCCACGTCTTCCAGAAAATCCATGTCTTCCAGAAAATCCATGTCTTCCAGTAACCTCCCAGTCTTCCAGAAAATCCACGTCTTCCCAACAATCCAAGTCTTCCGGATAATTTGGGTCTTCCTGAAAATCTACGTCTTCCAAAAAAGCCATGTCTTCCAGAAAATCCACATCTTCCAATGGCCTCCAGGTCTTCCAGACTATCCATGTCTTCCAGAAAATCCTTGTCTTCCCTTAAATCTATAGCTTCCAAAAAATCCGGGTCTTCCAGGAAATCCGTGTCTTCCAGCAAGTCCACGTCTTCCAACAAAGCCATGTCTTCCAGACTATCCATGTCTTCCAGAAAATCCTTGTCTTCCCTCAAATCCATAGCTTCCGAAAAATCCAGGTCTTCCAGGAAATCCGTGTCTTCCAGCAAATCCACGTCTTCCAACAAAGCCATGTCTTCCATCAAATTAATGTCTTCCAGCCTACTTGTGTCTTCCAACAAAGGTACGTCTTCCAACAAAGGTACGTCTTCCAACAAAGGTATGTCTTCCAACAAAGGTACGTCTTCCAGAAAATCCACGTCTTCCAACCAAGCCATGTCTTCCAGAAAATCCACGTCTTCCAGAAAATATATGTCTTCCAACTAAGCTACGTCTTCCAACAAATC
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CATGTCTTCCTATATCTCCAGGTCTTCCAGCATCTCCAGGGCTTCCAGCATCTGCTCGTCTTCCAACATCTCCACGTCTTCCAGCATCTCTGTGTCTTCCAGCATCTTCATGTCTTCCAACAACTACCCAGTCTTCCATCAACTGGCTCAATATCCATGTCTTCCAACGTCTCCAGTGTGCTGATCTTCTGACATTCAGGTCTTCCAGTGTCTGCAATATCCAG
pcDNA3.1(+) ZKSCAN1 MCS Vector + HIPK3 ExonUsed in Supplemental Figure 6Bgtatggcctcacaagtcttggtctacccaccatatgtttatcaaactcagtcaagtgccttttgtagtgtgaagaaactcaaagtagagccaagcagttgtgtattccaggaaagaaactatccacggacctatgtgaatggtagaaactttggaaattctcatcctcccactaagggtagtgcttttcagacaaagataccatttaatagacctcgaggacacaacttttcattgcagacaagtgctgttgttttgaaaaacactgcaggtgctacaaaggtcatagcagctcaggcacagcaagctcacgtgcaggcacctcagattggggcgtggcgaaacagattgcatttcctagaaggcccccagcgatgtggattgaagcgcaagagtgaggagttggataatcatagcagcgcaatgcagattgtcgatgaattgtccatacttcctgcaatgttgcaaaccaacatgggaaatccagtgacagttgtgacagctaccacaggatcaaaacagaattgtaccactggagaaggtgactatcagttagtacagcatgaagtcttatgctccatgaaaaatacttacgaagtccttgattttcttggtcgaggcacgtttggccaggtagttaaatgctggaaaagagggacaaatgaaattgtagcaatcaaaattttgaagaatcatccttcttatgcccgtcaaggtcaaatagaagtgagcatattagcaaggctcagtactgaaaatgctgatgaatataactttgtacgagcttatgaatgctttcagcaccgtaaccatacttgtttagtctttgagatgctggaacaaaacttgtatgactttctgaaacaaaataaatttagtcccctgccactaaaagtgattcggcccattcttcaacaagtggccactgcactgaaaaaattgaaaagtcttggtttaattcatgctgatctcaagccagagaatattatgttggtggatcctgttcggcagccttacagggttaaagtaatagactttgggtcggccagtcatgtatcaaagactgtttgttcaacatatctacaatctcggtactacag
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The following plasmids were generated by inserting the indicated sequences into the EcoRV and PacI sites of pcDNA3.1(+) ZKSCAN1 MCS Vector.
pcDNA3.1(+) ZKSCAN1 MCS Vector + ZKSCAN1 548-847 EcoRV-PacIUsed in Supplemental Figure 6FGAATAGTAAAGAAACACATCATAAAACCTCCCAGGACATAAAGGTGAGCACAGACCCTGTTTGGATCAAGTCAGTTCCTGGAGCCTGAATGATGACTGCTGAATCACGGGAAGCCACGGGTCTGTCCCCACAGGCTGCACAGGAGAAGGATGGTATCGTAATAGTGAAGGTGGAAGAGGAAGATGAGGAAGACCACATGTGGGGGCAGGATTCCACCCTACAGGACACGCCTCCTCCAGACCCAGAGATATTCCGCCAACGCTTCAGGCGCTTCTGTTACCAGAACACTTTTGGGCCCCG
pcDNA3.1(+) ZKSCAN1 MCS Vector + ZKSCAN1 548-1047 EcoRV-PacIUsed in Supplemental Figure 6FGAATAGTAAAGAAACACATCATAAAACCTCCCAGGACATAAAGGTGAGCACAGACCCTGTTTGGATCAAGTCAGTTCCTGGAGCCTGAATGATGACTGCTGAATCACGGGAAGCCACGGGTCTGTCCCCACAGGCTGCACAGGAGAAGGATGGTATCGTAATAGTGAAGGTGGAAGAGGAAGATGAGGAAGACCACATGTGGGGGCAGGATTCCACCCTACAGGACACGCCTCCTCCAGACCCAGAGATATTCCGCCAACGCTTCAGGCGCTTCTGTTACCAGAACACTTTTGGGCCCCGAGAGGCTCTCAGTCGGCTGAAGGAACTTTGTCATCAGTGGCTGCGGCCAGAAATAAACACCAAGGAACAGATCCTGGAGCTTCTGGTGCTAGAGCAGTTTCTTTCCATCCTGCCCAAGGAGCTCCAGGTCTGGCTGCAGGAATACCGCCCCGATAGTGGAGAGGAGGCCGTGACCCTTCTAGAAGACTTGGAGCTTGATT
pcDNA3.1(+) ZKSCAN1 MCS Vector + ZKSCAN1 548-1417 (No Intron) EcoRV-PacIUsed in Supplemental Figure 6FGAATAGTAAAGAAACACATCATAAAACCTCCCAGGACATAAAGgtgagcacagaccctgtttggatcaagtcagttcctggagcctgaatgatgactgctgaatcacgggaagccacgggtctgtccccacagGCTGCACAGGAGAAGGATGGTATCGTAATAGTGAAGGTGGAAGAGGAAGATGAGGAAGACCACATGTGGGGGCAGGATTCCACCCTACAGGACACGCCTCCTCCAGACCCAGAGATATTCCGCCAACGCTTCAGGCGCTTCTGTTACCAGAACACTTTTGGGCCCCGAGAGGCTCTCAGTCGGCTGAAGGAACTTTGTCATCAGTGGCTGCGGCCAGAAATAAACACCAAGGAACAGATCCTGGAGCTTCTGGTGCTAGAGCAGTTTCTTTCCATCCTGCCCAAGGAGCTCCAGGTCTGGCTGCAGGAATACCGCCCCGATAGTGGAGAGGAGGCCGTGACCCTTCTAGAAGACTTGGAGCTTGATTTATCAGGACAACAGGTCCCAGGTCAAGTTCATGGACCTGAGATGCTCGCAAGGGGGATGGTGCCTCTGGATCCAGTTCAGGAGTCCTCGAGCTTTGACCTTCATCACGAGGCCACCCAGTCCCACTTCAAACATTCGTCTCGGAAACCC
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To generate mammalian Laccase2 expression plasmids terminating in various 3’ end formation sequences, the previously described pCRII-TOPO CMV-cGFP-mMALAT1_3’ plasmid (Wilusz et al. 2012) was modified. The vector was first cut with NotI and the following sequence (comprised of a SalI restriction site followed by the 174-nt mMALAT1_3’ region) was inserted:
GTCGACgattcgtcagtagggttgtaaaggtttttcttttcctgagaaaacaaccttttgttttctcaggttttgctttttggcctttccctagctttaaaaaaaaaaaagcaaaagacgctggtggctggcactcctggtttccaggacggggttcaagtccctgcggtgtctttgctt
The cGFP ORF was then replaced by nucleotides 100-1945 of Laccase2 (sequence below), which were inserted in between the EcoRV and SalI restriction sites,
TGCGAAGCCAGTAAGCTACAAAAAGAAAaaacaagagagaatgctatagtcgtatagtatagtttcccgactatctgatacccattacttatctagggggaatgcgaacccaaaattttatcagttttctcggatatcgatagatattggggaataaatttaaataaataaattttgggcgggtttagggcgtggcaaaaagttttttggcaaatcgctagaaatttacaagacttataaaattatgaaaaaatacaacaaaattttaaacacgtgggcgtgacagttttggacggttttagggcgttagagtaggcgaggacagggttacatcgactaggctttgatcctgatcaagaatatatatactttataccgcttccttctacatgttacctatttttcaacgaatctagtatacctttttactgtacgatttatgggtataaTAATAAGCTAAATCGAGACTAAGttttattgttatatatattttttttattttatGCAGAAAATCAACCAGATCTTACACCAATGGGGTCCTTCTGGCAGCAACATGTGGCCACTCCTAGTAGTTCAACTCCATTCGATTCACCATTTTTTTCGGCAACACACGGCCTTGTGCAGACTCACCCGTCTATAGGTGGAAGCCACTCTGGGCCCGCTCCATTAAATGGCAATAGCCTTGGGTCTGGAGCTCCATCCTCAATCCTAGCTGGTACTAAGCCAGCCCTAACCGCTGGAAGCGGTTACCTAAACTCACGCGGGAGTCTACCGACCTCTGCACGTTATCCAGCTAATAAGATCACAGGAACCGTCGTGGAACCCAACCCAAAATCTCCATTCCGCCACTTAGATTTCTCTACTAGTGCAACTGCCGAGCTGCGGCGCAATCCTGCGCTATCTGCTCCTGATGAATGTGCACGCGCTTGTCGCGAAGGCGAGCCTCCGAGAATTTGCTACTATCATTTTACGTTGGAATACTACACAGTATTGGGAGCGTAAGTATTCAAAATTCCAAAATTTTTTACTAGAAATATTCGATTTTTTAATAGGCAGTTTCTATACTATTGTATACTATTGtagattcgttgaaaagtatgtaacaggaagaataaagcatttccgaccatgtaaagtatatatattcttaataaggatcaatagccgagtcgatctcgccatgtccgtctgtcttattattttattaccgccgagacatcaggaactataaaagctagaaggatgagttttagcatacagattctagagacaaggacgcagagcaagtttgttgatccatgctgccacgctttaactttctcaaattgcccaaaactgccatgcccacatttttgaactattttcgaaattttttcataattgtattactcgtgtaaatttccatcaatttgccaaaaaactttttgtcacgcgttaacgccctaaagccgccaatttggtcacgcccacactattgaacaattatcaaattttttctcattttattccccaatatctatcgatatccccgattatgaaattattaaatttcgcgttcgcattcacactagctgagtaacgagtatctgatagttggggaaatcgactTATTTTTTATATACAATGAAAATGAATTTAATCATATGAATATCGATTATAGCTTTTTATTTAATATGAATATTTATTTGGGCTTAAGGTGTAACCTcctcgacataagactcacatggcgcaggcacattgaagacaaaaatactcaTTGTCGGGTCTCGCACCCTCCAGCAGCACCTAAAATTATGTCTTCAATTATTGCCAACATTGGAGACACAATTAGTCTGTGGCACCTCAG
Finally, the various 3’ end formation sequences below were then inserted in between the SalI and NotI restriction sites.
pCRII-TOPO Laccase2 100-1945 - SV40 pA SenseUsed in Figure 5Caacttgtttattgcagcttataatggttacaaataaagcaatagcatcacaaatttcacaaataaagcatttttttcactgcattctagttgtggtttgtccaaactcatcaatgtatctta
pCRII-TOPO Laccase2 100-1945 - SV40 pA AntisenseUsed in Figure 5Ctaagatacattgatgagtttggacaaaccacaactagaatgcagtgaaaaaaatgctttatttgtgaaatttgtgatgctattgctttatttgtaaccattataagctgcaataaacaagtt
pCRII-TOPO Laccase2 100-1945 - mMALAT1_3’ SenseUsed in Figure 5Cgattcgtcagtagggttgtaaaggtttttcttttcctgagaaaacaaccttttgttttctcaggttttgctttttggcctttccctagctttaaaaaaaaaaaagcaaaagacgctggtggctggcactcctggtttccaggacggggttcaagtccctgcggtgtctttgctt
pCRII-TOPO Laccase2 100-1945 - mMALAT1_3’ AntisenseUsed in Figure 5Caagcaaagacaccgcagggacttgaaccccgtcctggaaaccaggagtgccagccaccagcgtcttttgcttttttttttttaaagctagggaaaggccaaaaagcaaaacctgagaaaacaaaaggttgttttctcaggaaaagaaaaacctttacaaccctactgacgaatc
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To generate ZKSCAN1 expression plasmids terminating in various 3’ end formation sequences, the previously described pCRII-TOPO CMV-cGFP-mMALAT1_3’ plasmid (Wilusz et al. 2012) was modified. The vector was first cut with NotI and the following sequence (comprised of a SalI restriction site followed by the 174-nt mMALAT1_3’ region) was inserted:
GTCGACgattcgtcagtagggttgtaaaggtttttcttttcctgagaaaacaaccttttgttttctcaggttttgctttttggcctttccctagctttaaaaaaaaaaaagcaaaagacgctggtggctggcactcctggtttccaggacggggttcaagtccctgcggtgtctttgctt
The cGFP ORF was then replaced by nucleotides 1-2232 of ZKSCAN1 (sequence below), which were inserted in between the EcoRV and SalI restriction sites,
TGCTGTCAACCTGCAGGTAAATCACTCTGACTCCAAGCAAACACCCTGAGGGTGGAGTCGGCCTGTTTGAGGTTTCTGTTTGCAAATCTTGATATATGAAGTGACAGTGGAGATTGTACAGTTTTTTCCTCGATTTGTCAGGAtttttttttttttgacggagtttaacttcttgtctcccaggtaggaagtgcagtggcgtaatctcggctcactacaacctccacctcctgggttcaagcgtttctcctgcctcagctttccgagtagctgggattacaggcgcctgccaccatgccctgctgacttttgtatttttagtagagacggggtttcaccatgttggccaggctggtcttgaactcctgaccgcaggcgattggcctgcctcggcctcccaaagtgctgagattacaggcgtgagccaccacccccggccTCAGGAGCGTTCTGATAGTGCCTCGATGTGCTGCCTCCTATAAAGTGTTAGCAGCACAGATCACTTTTTGTAAAGGTACGTACTAATGACTTTTTTTTTATACTTCAGGAATAGTAAAGAAACACATCATAAAACCTCCCAGGACATAAAGGTGAGCACAGACCCTGTTTGGATCAAGTCAGTTCCTGGAGCCTGAATGATGACTGCTGAATCACGGGAAGCCACGGGTCTGTCCCCACAGGCTGCACAGGAGAAGGATGGTATCGTAATAGTGAAGGTGGAAGAGGAAGATGAGGAAGACCACATGTGGGGGCAGGATTCCACCCTACAGGACACGCCTCCTCCAGACCCAGAGATATTCCGCCAACGCTTCAGGCGCTTCTGTTACCAGAACACTTTTGGGCCCCGAGAGGCTCTCAGTCGGCTGAAGGAACTTTGTCATCAGTGGCTGCGGCCAGAAATAAACACCAAGGAACAGATCCTGGAGCTTCTGGTGCTAGAGCAGTTTCTTTCCATCCTGCCCAAGGAGCTCCAGGTCTGGCTGCAGGAATACCGCCCCGATAGTGGAGAGGAGGCCGTGACCCTTCTAGAAGACTTGGAGCTTGATTTATCAGGACAACAGGTAAAAAGAGGTGAAACCTATTATGTGTGAGCAGGGCACAGACGTTGAAACTGGAGCCAGGAGAAGTATTGGCAGGCTTTAGGTTATTAGGTGGTTACTCTGTCTTAAAAATGTTCTGGCTTTCTTCCTGCATCCACTGGCATACTCATGGTCTGTTTTTAAATATTTTAATTCCCATTTACAAAGTGATTTACCCACAAGCCCAACCTGTCTGTCTTCAGGTCCCAGGTCAAGTTCATGGACCTGAGATGCTCGCAAGGGGGATGGTGCCTCTGGATCCAGTTCAGGAGTCCTCGAGCTTTGACCTTCATCACGAGGCCACCCAGTCCCACTTCAAACATTCGTCTCGGAAACCCCGCCTCTTACAGTCACGAGGTAAGAAGCAAGGTTTCATTTAGGGGAAGGGAAATGATTCAGGACGAGAGTCTTTGTGCTGCTGAGTGCCTGTGATGAAGAAGCATGTTAGTcctgggcaacgtagcgagaccccatctctacaaaaaatagaaaaattagccaggtatagtggcgcacacctgtgattccagctacgcaggaggctgaggtgggaggattgcttgagcccaggaggttgaggctgcagtgagctgtaatcatgccactactccaacctgggcaacacagcaaggaccctgtctcaaaaGCTACTTACAGAAAAGAATTAggctcggcacggtagctcacacctgtaatcccagcactttgggaggctgaggcgggcagatcacttgaggtcaggagtttgagaccagcctggccaacatggtgaaaccttgtctctactaaaaatatgaaaattagccaggcatggtggcacattcctgtaatcccagctactcgggaggctgaggcaggagaatcacttgaacccaggaggtggaggttgcagtaagccgagatcgtaccactgtgctctagccttggtgacagagcgagactgtcttaaaaaaaaaaaaaaaaaaaaaagaattaattaaaaatttaaaaaaaaatgaaaaaaaGCTGCATGCTTGTTTTTTGTTTTTAGTTATTCTACATTGTTGTCATTATTACCAAATATTGGGGAAAATACAACTTACAGACCAATCTCAGGAGTTAAATGTTACTACGAAGGCAAATGAACTATGCGTAATGAACCTGGTAGGCATTA
Finally, the various 3’ end formation sequences below were then inserted in between the SalI and NotI restriction sites.
pCRII-TOPO ZKSCAN1 1-2232 - SV40 pA SenseUsed in Figure 5Eaacttgtttattgcagcttataatggttacaaataaagcaatagcatcacaaatttcacaaataaagcatttttttcactgcattctagttgtggtttgtccaaactcatcaatgtatctta
pCRII-TOPO ZKSCAN1 1-2232 - SV40 pA AntisenseUsed in Figure 5Etaagatacattgatgagtttggacaaaccacaactagaatgcagtgaaaaaaatgctttatttgtgaaatttgtgatgctattgctttatttgtaaccattataagctgcaataaacaagtt
pCRII-TOPO ZKSCAN1 1-2232 - mMALAT1_3’ SenseUsed in Figure 5Egattcgtcagtagggttgtaaaggtttttcttttcctgagaaaacaaccttttgttttctcaggttttgctttttggcctttccctagctttaaaaaaaaaaaagcaaaagacgctggtggctggcactcctggtttccaggacggggttcaagtccctgcggtgtctttgctt
pCRII-TOPO ZKSCAN1 1-2232 - mMALAT1_3’ AntisenseUsed in Figure 5Eaagcaaagacaccgcagggacttgaaccccgtcctggaaaccaggagtgccagccaccagcgtcttttgcttttttttttttaaagctagggaaaggccaaaaagcaaaacctgagaaaacaaaaggttgttttctcaggaaaagaaaaacctttacaaccctactgacgaatc
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The following plasmids were previously described:
pcDNA3.1(+) CircRNA Mini VectorUsed in Figure 7CUsed in Supplemental Figure 6BPreviously described in Liang and Wilusz (2014) Genes Dev.
pcDNA3.1(+) CircRNA Mini Vector + ZKSCAN1 548-847Used in Figure 7CPreviously described in Liang and Wilusz (2014) Genes Dev.
pcDNA3.1(+) CircRNA Mini Vector + ZKSCAN1 548-1047Used in Figure 7CPreviously described in Liang and Wilusz (2014) Genes Dev.
pcDNA3.1(+) CircRNA Mini Vector + ZKSCAN1 548-1417 (No Intron)Used in Figure 7CPreviously described in Liang and Wilusz (2014) Genes Dev.
pcDNA3.1(+) CircRNA Mini Vector + HIPK3 ExonUsed in Supplemental Figure 6BPreviously described in Liang and Wilusz (2014) Genes Dev.
pCRII-TOPO ZKSCAN1 400-1782 - SV40 pA SenseUsed in Figure 5EPreviously described in Liang and Wilusz (2014) Genes Dev.
pCRII-TOPO ZKSCAN1 400-1782 - SV40 pA AntisenseUsed in Figure 5EPreviously described in Liang and Wilusz (2014) Genes Dev.
pCRII-TOPO ZKSCAN1 400-1782 - mMALAT1_3’ SenseUsed in Figure 5EPreviously described in Liang and Wilusz (2014) Genes Dev.
pCRII-TOPO ZKSCAN1 400-1782 - mMALAT1_3’ AntisenseUsed in Figure 5EPreviously described in Liang and Wilusz (2014) Genes Dev.
pcDNA3-ciRS7Used in Figure 7DPreviously described in Hansen et al. (2013) Nature