5' GAT CTC GAT CCC GCG AAA TTA ATA CGA CTC ACT ATA GGG … · Wednesday, November 9, 2011 11:30 AM...
Transcript of 5' GAT CTC GAT CCC GCG AAA TTA ATA CGA CTC ACT ATA GGG … · Wednesday, November 9, 2011 11:30 AM...
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Wednesday, November 9, 2011 11:30 AM Page 1 of 7HisPhnD90.L1ADÆÆ
T7 promoter
GAT CTC GAT CCC GCG AAA TTA ATA CGA CTC ACT ATA GGG AGA CCA CAA CGG TTT5' 54
NdeI
M H H
-9 -8 -7
CCC TCT AGA AAT AAT TTT GTT TAA CTT TAA GAA GGA GAT ATA CAT ATG CAT CAT5' 108
BamHI
PhnD-NHalf
H H H H G S E E Q E K A L N F G I I
-6 -5 -4 -3 -2 -1 1 2 3 4 5 6 7 8 9 10 11 12
CAC CAC CAC CAT GGA TCC GAA GAA CAA GAA AAA GCT CTG AAC TTC GGT ATC ATC5' 162
PstI
PhnD-NHalf
S T E S Q Q N L K P Q W T P F L Q D
13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30
TCC ACT GAA AGC CAA CAG AAC CTG AAG CCG CAG TGG ACC CCG TTC CTG CAG GAT5' 216
BspEI
PhnD-NHalf
M E K K L G V K V N A F F A P D Y A
31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48
ATG GAG AAG AAA CTG GGC GTT AAA GTC AAC GCT TTC TTC GCT CCG GAC TAC GCA5' 270
PhnD-NHalf
G I I Q G M R F N K V D I A W Y G N
49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66
GGC ATC ATC CAG GGT ATG CGT TTC AAC AAA GTC GAT ATC GCC TGG TAC GGC AAT5' 324
PhnD-NHalf
L S A M E A V D R A N G Q V F A Q T
67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84
CTG TCT GCA ATG GAA GCG GTC GAT CGT GCC AAC GGT CAG GTG TTC GCG CAG ACG5' 378
PhnD-NHalf cpGFP
V A A D
85 86 87 88
A D N V Y I M A D K Q R N G
147 148 149 150 151 152 153 154 155 156 157 158 159 160
GTT GCA GCG GAC gcg gac AAC GTC TAT ATC ATG GCC GAC AAG CAG AgG AAC GGC5' 432
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Wednesday, November 9, 2011 11:30 AM Page 2 of 7HisPhnD90.L1ADÆÆ
cpGFP
I K A N F K I R H N I E D G G V Q L
161 162 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178
ATC AAG GCG AAC TTC AAG ATC CGC CAC AAC ATC GAG GAC GGC GGC GTG CAG CTC5' 486
cpGFP
A Y H Y Q Q N T P I G D G P V L L P
179 180 181 182 183 184 185 186 187 188 189 190 191 192 193 194 195 196
GCC TAC CAC TAC CAG CAG AAC ACC CCC ATC GGC GAC GGC CCC GTG CTG CTG CCC5' 540
cpGFP
D N H Y L S T Q S K L S K D P N E K
197 198 199 200 201 202 203 204 205 206 207 208 209 210 211 212 213 214
GAC AAC CAC TAC CTG AGC ACC CAG TCC AAA CTT TCG AAA GAC CCC AAC GAG AAG5' 594
cpGFP
R D H M V L L E F V T A A G I T L G
215 216 217 218 219 220 221 222 223 224 225 226 227 228 229 230 231 232
CGC GAT CAC ATG GTC CTG CTG GAG TTC GTG ACC GCC GCC GGG ATC ACT CTC GGC5' 648
cpGFP
M D E L Y K
233 234 235 236 237 238
G G T G G S
1 2 3 4 5 6
M V S K G E
1 2 3 4 5 6
ATG GAC GAG CTG TAC AAG GGC GGT ACC GGA GGG AGC ATG GTG AGC AAG GGC GAG5' 702
cpGFP
E L F T G V V P I L V E L D G D V N
7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
GAG CTG TTC ACC GGG GTG GTG CCC ATC CTG GTC GAG CTG GAC GGC GAC GTA AAC5' 756
cpGFP
G H K F S V S G E G E G D A T Y G K
25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42
GGC CAC AAG TTC AGC GTG TCC GGC GAG GGC GAG GGC GAT GCC ACC TAC GGC AAG5' 810
cpGFP
L T L K F I C T T G K L P V P W P T
43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
CTG ACC CTG AAG TTC ATC TGC ACC ACC GGC AAG CTG CCC GTG CCC TGG CCC ACC5' 864
cpGFP
L V T T L T Y G V Q C F S R Y P D H
61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78
CTC GTG ACC ACC CTG ACC TAC GGC GTG CAG TGC TTC AGC CGC TAC CCC GAC CAC5' 918
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Wednesday, November 9, 2011 11:30 AM Page 3 of 7HisPhnD90.L1ADÆÆ
cpGFP
M K Q H D F F K S A M P E G Y I Q E
79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96
ATG AAG CAG CAC GAC TTC TTC AAG TCC GCC ATG CCC GAA GGC TAC ATC CAG GAG5' 972
cpGFP
R T I F F K D D G N Y K T R A E V K
97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114
CGC ACC ATC TTC TTC AAG GAC GAC GGC AAC TAC AAG ACC CGC GCC GAG GTG AAG5' 1026
cpGFP
F E G D T L V N R I E L K G I D F K
115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132
TTC GAG GGC GAC ACC CTG GTG AAC CGC ATC GAG CTG AAG GGC ATC GAC TTC AAG5' 1080
PhnD-CHalfcpGFP
E D G N I L G H K L E Y N F N
133 134 135 136 137 138 139 140 141 142 143 144 145 146 147
P G Y
91 92 93
GAG GAC GGC AAC ATC CTG GGG CAC AAG CTG GAG TAC AAC TTT AAC CCG GGC TAT5' 1134
PhnD-CHalf
W S V L I V N K D S P I N N L N D L
94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111
TGG TCC GTG CTG ATC GTT AAC AAA GAC AGC CCG ATT AAC AAT CTG AAC GAC CTG5' 1188
PhnD-CHalf
L A K R K D L T F G N G D P N S T S
112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129
CTG GCA AAA CGC AAG GAC CTG ACC TTC GGC AAC GGT GAT CCT AAC TCC ACG TCT5' 1242
PhnD-CHalf
G F L V P G Y Y V F A K N N I S A S
130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147
GGC TTC CTG GTG CCA GGT TAC TAC GTC TTC GCC AAA AAC AAC ATC TCC GCC TCT5' 1296
PhnD-CHalf
D F K R T V N A G H E T N A L A V A
148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165
GAC TTC AAA CGT ACC GTA AAC GCA GGT CAC GAA ACC AAC GCT CTG GCT GTT GCT5' 1350
PhnD-CHalf
N K Q V D V A T N N T E N L D K L K
166 167 168 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183
AAC AAG CAG GTG GAC GTG GCG ACT AAT AAC ACG GAA AAC CTG GAC AAG CTG AAA5' 1404
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Wednesday, November 9, 2011 11:30 AM Page 4 of 7HisPhnD90.L1ADÆÆ
BspEI
PhnD-CHalf
T S A P E K L K E L K V I W K S P L
184 185 186 187 188 189 190 191 192 193 194 195 196 197 198 199 200 201
ACC AGC GCT CCG GAA AAA CTG AAA GAA CTG AAA GTA ATC TGG AAA AGC CCG CTG5' 1458
PhnD-CHalf
I P G D P I V W R K N L S E T T K D
202 203 204 205 206 207 208 209 210 211 212 213 214 215 216 217 218 219
ATC CCG GGT GAC CCT ATC GTA TGG CGT AAA AAC CTG TCC GAA ACC ACT AAA GAC5' 1512
PhnD-CHalf
K I Y D F F M N Y G K T P E E K A V
220 221 222 223 224 225 226 227 228 229 230 231 232 233 234 235 236 237
AAA ATT TAT GAC TTT TTC ATG AAC TAC GGT AAA ACC CCG GAA GAA AAG GCT GTG5' 1566
PstI
PhnD-CHalf
L E R L G W A P F R A S S D L Q L V
238 239 240 241 242 243 244 245 246 247 248 249 250 251 252 253 254 255
CTG GAA CGT CTG GGC TGG GCT CCG TTC CGT GCT TCC AGC GAC CTG CAG CTG GTT5' 1620
PhnD-CHalf
P I R Q L A L F K E M Q S V K D N K
256 257 258 259 260 261 262 263 264 265 266 267 268 269 270 271 272 273
CCA ATC CGC CAG CTG GCT CTG TTC AAA GAA ATG CAG TCT GTT AAA GAT AAC AAG5' 1674
PhnD-CHalf
G L N E Q D K L A K T T A I Q A Q L
274 275 276 277 278 279 280 281 282 283 284 285 286 287 288 289 290 291
GGT CTG AAC GAA CAG GAC AAG CTG GCT AAG ACT ACT GCG ATC CAG GCA CAG CTG5' 1728
PhnD-CHalf
D D L D R R N N A R S A M S S V S K
292 293 294 295 296 297 298 299 300 301 302 303 304 305 306 307 308 309
GAC GAT CTG GAT CGT agg AAT AAT GCA agg AGC GCA ATG TCT TCC GTT AGC AAA5' 1782
HindIIIEcoRI
PhnD-CHalf
A V Q .
310 311 312 313
GCG GTC CAG taa GAA TTC GAA GCT TGA TCC GGC TGC TAA CAA AGC CCG AAA GGA5' 1836
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Wednesday, November 9, 2011 11:30 AM Page 5 of 7HisPhnD90.L1ADÆÆ
AGC TGA GTT GGC TGC TGC CAC CGC TGA GCA ATA ACT AGC ATA ACC CCT TGG GGC5' 1890
BspEI
CTC TAA ACG GGT CTT GAG GGG TTT TTT GCT GAA AGG AGG AAC TAT ATC CGG ATC5' 1944
TGG CGT AAT AGC GAA GAG GCC CGC ACC GAT CGC CCT TCC CAA CAG TTG CGC AGC5' 1998
f1 origin
CTG AAT GGC GAA TGG GAC GCG CCC TGT AGC GGC GCA TTA AGC GCG GCG GGT GTG5' 2052
f1 origin
GTG GTT ACG CGC AGC GTG ACC GCT ACA CTT GCC AGC GCC CTA GCG CCC GCT CCT5' 2106
f1 origin
TTC GCT TTC TTC CCT TCC TTT CTC GCC ACG TTC GCC GGC TTT CCC CGT CAA GCT5' 2160
f1 origin
CTA AAT CGG GGG CTC CCT TTA GGG TTC CGA TTT AGT GCT TTA CGG CAC CTC GAC5' 2214
f1 origin
CCC AAA AAA CTT GAT TAG GGT GAT GGT TCA CGT AGT GGG CCA TCG CCC TGA TAG5' 2268
f1 origin
ACG GTT TTT CGC CCT TTG ACG TTG GAG TCC ACG TTC TTT AAT AGT GGA CTC TTG5' 2322
f1 origin
TTC CAA ACT GGA ACA ACA CTC AAC CCT ATC TCG GTC TAT TCT TTT GAT TTA TAA5' 2376
f1 origin
GGG ATT TTG CCG ATT TCG GCC TAT TGG TTA AAA AAT GAG CTG ATT TAA CAA AAA5' 2430
f1 origin
TTT AAC GCG AAT TTT AAC AAA ATA TTA ACG CTT ACA ATT TAG GTG GCA CTT TTC5' 2484
GGG GAA ATG TGC GCG GAA CCC CTA TTT GTT TAT TTT TCT AAA TAC ATT CAA ATA5' 2538
TGT ATC CGC TCA TGA GAC AAT AAC CCT GAT AAA TGC TTC AAT AAT ATT GAA AAA5' 2592
GGA AGA GTA TGA GTA TTC AAC ATT TCC GTG TCG CCC TTA TTC CCT TTT TTG CGG5' 2646
CAT TTT GCC TTC CTG TTT TTG CTC ACC CAG AAA CGC TGG TGA AAG TAA AAG ATG5' 2700
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CTG AAG ATC AGT TGG GTG CAC GAG TGG GTT ACA TCG AAC TGG ATC TCA ACA GCG5' 2754
GTA AGA TCC TTG AGA GTT TTC GCC CCG AAG AAC GTT TTC CAA TGA TGA GCA CTT5' 2808
TTA AAG TTC TGC TAT GTG GCG CGG TAT TAT CCC GTA TTG ACG CCG GGC AAG AGC5' 2862
AAC TCG GTC GCC GCA TAC ACT ATT CTC AGA ATG ACT TGG TTG AGT ACT CAC CAG5' 2916
TCA CAG AAA AGC ATC TTA CGG ATG GCA TGA CAG TAA GAG AAT TAT GCA GTG CTG5' 2970
CCA TAA CCA TGA GTG ATA ACA CTG CGG CCA ACT TAC TTC TGA CAA CGA TCG GAG5' 3024
GAC CGA AGG AGC TAA CCG CTT TTT TGC ACA ACA TGG GGG ATC ATG TAA CTC GCC5' 3078
TTG ATC GTT GGG AAC CGG AGC TGA ATG AAG CCA TAC CAA ACG ACG AGC GTG ACA5' 3132
CCA CGA TGC CTG TAG CAA TGG CAA CAA CGT TGC GCA AAC TAT TAA CTG GCG AAC5' 3186
TAC TTA CTC TAG CTT CCC GGC AAC AAT TAA TAG ACT GGA TGG AGG CGG ATA AAG5' 3240
TTG CAG GAC CAC TTC TGC GCT CGG CCC TTC CGG CTG GCT GGT TTA TTG CTG ATA5' 3294
AAT CTG GAG CCG GTG AGC GTG GGT CTC GCG GTA TCA TTG CAG CAC TGG GGC CAG5' 3348
ATG GTA AGC CCT CCC GTA TCG TAG TTA TCT ACA CGA CGG GGA GTC AGG CAA CTA5' 3402
TGG ATG AAC GAA ATA GAC AGA TCG CTG AGA TAG GTG CCT CAC TGA TTA AGC ATT5' 3456
GGT AAC TGT CAG ACC AAG TTT ACT CAT ATA TAC TTT AGA TTG ATT TAA AAC TTC5' 3510
ATT TTT AAT TTA AAA GGA TCT AGG TGA AGA TCC TTT TTG ATA ATC TCA TGA CCA5' 3564
AAA TCC CTT AAC GTG AGT TTT CGT TCC ACT GAG CGT CAG ACC CCG TAG AAA AGA5' 3618
TCA AAG GAT CTT CTT GAG ATC CTT TTT TTC TGC GCG TAA TCT GCT GCT TGC AAA5' 3672
CAA AAA AAC CAC CGC TAC CAG CGG TGG TTT GTT TGC CGG ATC AAG AGC TAC CAA5' 3726
CTC TTT TTC CGA AGG TAA CTG GCT TCA GCA GAG CGC AGA TAC CAA ATA CTG TTC5' 3780
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Wednesday, November 9, 2011 11:30 AM Page 7 of 7HisPhnD90.L1ADÆÆ
TTC TAG TGT AGC CGT AGT TAG GCC ACC ACT TCA AGA ACT CTG TAG CAC CGC CTA5' 3834
CAT ACC TCG CTC TGC TAA TCC TGT TAC CAG TGG CTG CTG CCA GTG GCG ATA AGT5' 3888
CGT GTC TTA CCG GGT TGG ACT CAA GAC GAT AGT TAC CGG ATA AGG CGC AGC GGT5' 3942
CGG GCT GAA CGG GGG GTT CGT GCA CAC AGC CCA GCT TGG AGC GAA CGA CCT ACA5' 3996
CCG AAC TGA GAT ACC TAC AGC GTG AGC TAT GAG AAA GCG CCA CGC TTC CCG AAG5' 4050
GGA GAA AGG CGG ACA GGT ATC CGG TAA GCG GCA GGG TCG GAA CAG GAG AGC GCA5' 4104
CGA GGG AGC TTC CAG GGG GAA ACG CCT GGT ATC TTT ATA GTC CTG TCG GGT TTC5' 4158
GCC ACC TCT GAC TTG AGC GTC GAT TTT TGT GAT GCT CGT CAG GGG GGC GGA GCC5' 4212
TAT GGA AAA ACG CCA GCA ACG CGG CCT TTT TAC GGT TCC TGG CCT TTT GCT GGC5' 4266
CTT TTG CTC ACA TGT TCT TTC CTG CGT TAT CCC CTG ATT CTG TGG ATA ACC GTA5' 4320
TTA CCG CCT TTG AGT GAG CTG ATA CCG CTC GCC GCA GCC GAA CGA CCG AGC GCA5' 4374
GCG AGT CAG TGA GCG AGG AAG CGG AAG AGC GCC CAA TAC GCA AAC CGC CTC TCC5' 4428
CCG CGC GTT GGC CGA TTC ATT AAT GCA G5' 4456