| 出品公司: | |
|---|---|
| 别名: | pET40b, pET 40b |
| 质粒类型: | 大肠杆菌蛋白表达 |
| 表达水平: | 高 |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 6190 bp |
| 5' 测序引物: | T7 |
| 5' 测序引物序列: | T7: 5'-TAATACGACTCACTATAGGG-3' |
| 载体标签: | His (中间和C端), N-DsbC, N-S, N-EK, N-Thrombin |
| 载体抗性: | Kanamycin |
| 备注: |
Encodes DsbC tag for export and periplasmic folding of target proteins.
|
| 表达宿主菌: | BL21(DE3) 、 Rosetta2(DE3)、C43(DE3) |
| 产品目录号: | ZY70091-3 |
| 稳定性: | 瞬时表达 Transient |
| 组成型: | 组成型 Constitutive |
| 病毒/非病毒: | 非病毒 |
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20℃低温保存
- 保质期：
三年
- 英文名：
pET-40b(+)
- 库存：
20
- 供应商：
泽叶生物
pET40b载体基本信息
pET40b载体质粒图谱和多克隆位点信息
| Feature Name | Start | End | |
|---|---|---|---|
| T7_terminator | 120 | 1 | |
| T7_Terminal_primer | 69 | 87 | |
| EK | 266 | 252 | |
| S15 | 326 | 282 | |
| 6xHIS | 386 | 369 | |
| T7_transl_en_RBS | 1140 | 1124 | |
| lacO | 1185 | 1158 | |
| T7_promoter | 1203 | 1185 | |
| tet (300 - 563) | 1239 | 1502 | |
| pBRrevBam_primer | 1310 | 1291 | |
| lacI | 1585 | 2676 | |
| tet (576 - 851) | 2735 | 3010 | |
| ROP | 3485 | 3676 | |
| pGEX_3_primer | 3692 | 3670 | |
| pBR322_origin | 4710 | 4091 | |
| KanR2 | 4816 | 5631 |
| ORF | Start | End | |
|---|---|---|---|
| ORF frame 3 | 1115 | 147 | |
| ORF frame 3 | 1116 | 1844 | |
| ORF frame 1 | 1717 | 2676 | |
| ORF frame 1 | 4816 | 5631 |
| Enzyme Name | Cut |
|---|---|
| XhoI | 174 |
| NotI | 182 |
| EagI | 182 |
| HindIII | 189 |
| SalI | 195 |
| SacI | 206 |
| EcoRI | 208 |
| BamHI | 214 |
| EcoRV | 222 |
| NcoI | 227 |
| KpnI | 271 |
| BglII | 274 |
| BstBI | 301 |
| SacII | 365 |
| SpeI | 396 |
| NdeI | 1114 |
| XbaI | 1152 |
| BclI | 1958 |
| ApaI | 2155 |
| HpaI | 2450 |
| FspI | 3026 |
| NruI | 4904 |
pET40b载体简介
The pET-40b(+) vector (Cat. No. 70091-3) is designed for expression of DsbC fusion proteins. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circle map. The cloning/ expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single stranded DNA that corresponds to the coding strand. Therefore, single stranded sequencing should be performed using the T7 terminator primer (Cat. No. 69337-3).
pET40b载体序列
LOCUS pET-40b(+) 6190 bp DNA Circular SYN
DEFINITION pET-40b(+)
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTOAUTHORAD1|Novagen Inc.|
FEATURES Location/Qualifiers
source 1..6190
/organism="pET-40b(+)"
/mol_type="other DNA"
terminator complement(1..120)
/label="T7_terminator"
misc_feature 69..87
/label="T7_Terminal_primer"
CDS complement(147..1115)
/label="ORF frame 3"
/translation="MKKGFMLFTLLAAFSGFAQADDAAIQQTLAKMGIKSSDIQPAPV
AGMKTVLTNSGVLYITDDGKHIIQGPMYDVSGTAPVNVTNKMLLKQLNALEKEMIVYK
APQEKHVITVFTDITCGYCHKLHEQMADYNALGITVRYLAFPRQGLDSDAEKEMKAIW
CAKDKNKAFDDVMAGKSVAPASCDVDIADHYALGVQLGVSGTPAVVLSNGTLVPGYQP
PKEMKEFLDEHQKMTSGKGSTSGSGHHHHHHSAGLVPRGSTAIGMKETAAAKFERQHM
DSPDLGTDDDDKSPGFSSTMAISDPNSSSVDKLAAALEHHHHHHHH*"
misc_feature complement(252..266)
/label="EK"
/translation="MKKGFMLFTLLAAFSGFAQADDAAIQQTLAKMGIKSSDIQPAPV
AGMKTVLTNSGVLYITDDGKHIIQGPMYDVSGTAPVNVTNKMLLKQLNALEKEMIVYK
APQEKHVITVFTDITCGYCHKLHEQMADYNALGITVRYLAFPRQGLDSDAEKEMKAIW
CAKDKNKAFDDVMAGKSVAPASCDVDIADHYALGVQLGVSGTPAVVLSNGTLVPGYQP
PKEMKEFLDEHQKMTSGKGSTSGSGHHHHHHSAGLVPRGSTAIGMKETAAAKFERQHM
DSPDLGTDDDDKSPGFSSTMAISDPNSSSVDKLAAALEHHHHHHHH*"
misc_feature complement(282..326)
/label="S15"
/translation="MKKGFMLFTLLAAFSGFAQADDAAIQQTLAKMGIKSSDIQPAPV
AGMKTVLTNSGVLYITDDGKHIIQGPMYDVSGTAPVNVTNKMLLKQLNALEKEMIVYK
APQEKHVITVFTDITCGYCHKLHEQMADYNALGITVRYLAFPRQGLDSDAEKEMKAIW
CAKDKNKAFDDVMAGKSVAPASCDVDIADHYALGVQLGVSGTPAVVLSNGTLVPGYQP
PKEMKEFLDEHQKMTSGKGSTSGSGHHHHHHSAGLVPRGSTAIGMKETAAAKFERQHM
DSPDLGTDDDDKSPGFSSTMAISDPNSSSVDKLAAALEHHHHHHHH*"
misc_feature complement(369..386)
/label="6xHIS"
/translation="MKKGFMLFTLLAAFSGFAQADDAAIQQTLAKMGIKSSDIQPAPV
AGMKTVLTNSGVLYITDDGKHIIQGPMYDVSGTAPVNVTNKMLLKQLNALEKEMIVYK
APQEKHVITVFTDITCGYCHKLHEQMADYNALGITVRYLAFPRQGLDSDAEKEMKAIW
CAKDKNKAFDDVMAGKSVAPASCDVDIADHYALGVQLGVSGTPAVVLSNGTLVPGYQP
PKEMKEFLDEHQKMTSGKGSTSGSGHHHHHHSAGLVPRGSTAIGMKETAAAKFERQHM
DSPDLGTDDDDKSPGFSSTMAISDPNSSSVDKLAAALEHHHHHHHH*"
CDS 1116..1844
/label="ORF frame 3"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
misc_feature complement(1124..1140)
/label="T7_transl_en_RBS"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
misc_feature complement(1158..1185)
/label="lacO"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
promoter complement(1185..1203)
/label="T7_promoter"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
gene 1239..1502
/label="tet (300 - 563)"
/gene="tet (300 - 563)"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
misc_feature complement(1291..1310)
/label="pBRrevBam_primer"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
misc_feature 1585..2676
/label="lacI"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
CDS 1717..2676
/label="ORF frame 1"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
gene 2735..3010
/label="tet (576 - 851)"
/gene="tet (576 - 851)"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
misc_feature 3485..3676
/label="ROP"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
misc_feature complement(3670..3692)
/label="pGEX_3_primer"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
rep_origin complement(4091..4710)
/label="pBR322_origin"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
gene 4816..5631
/label="KanR2"
/gene="KanR2"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
CDS 4816..5631
/label="ORF frame 1"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgtttagcag cctaggtatt aatcaattag tggtggtggt ggtggtggtg gtgctcgagt
181 gcggccgcaa gcttgtcgac ggagctcgaa ttcggatccg atatcgccat ggttgaggag
241 aagcccgggc tcttgtcgtc gtcatcggta cccagatctg ggctgtccat gtgctggcgt
301 tcgaatttag cagcagcggt ttctttcata ccaattgcag tactaccgcg tggcaccaga
361 cccgcggagt gatggtgatg gtgatgacca gaaccactag ttgatccttt accgctggtc
421 attttttggt gttcgtcgag aaattctttc atctctttcg gcggctggta acccggaaca
481 agtgtgccat tgctcagcac aactgccgga gtaccgctaa cgccaagctg gacgccaagt
541 gcgtaatggt cggcaatatc cacgtcgcaa ctggctggtg cgacgctttt acctgccatc
601 acatcatcaa acgctttgtt tttatctttc gcacaccaga tagctttcat ttctttctct
661 gcatcgctgt ccagcccctg gcgcgggaaa gcaagataac gcacggtgat ccccagcgcg
721 ttgtagtctg ccatttgctc atgcagtttg tggcagtaac cacaggtaat atcagtaaac
781 acggtgatga cgtgtttttc ctgcggcgct ttataaacga tcatctcttt ttcaagcgca
841 ttcaactgct ttaacagcat cttattggtg acattgaccg gagccgtgcc actaacgtca
901 tacattggcc cctgaatgat atgtttacca tcatcggtga tgtacaacac gccgctgtta
961 gtcagaactg tcttcatgcc agctacaggc gcgggctgaa tatcgctgct tttgatgccc
1021 attttggcta acgtttgttg aattgccgcg tcatcagcct gagcaaagcc tgaaaacgcc
1081 gctaacaaag taaacaacat aaaacctttc ttcatatgta tatctccttc ttaaagttaa
1141 acaaaattat ttctagaggg gaattgttat ccgctcacaa ttcccctata gtgagtcgta
1201 ttaatttcgc gggatcgaga tcgatctcga tcctctacgc cggacgcatc gtggccggca
1261 tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag
1321 atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc
1381 ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg
1441 tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag
1501 agcgtcgaga tcccggacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag
1561 cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc
1621 gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac
1681 gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc
1741 aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc
1801 agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa
1861 ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg
1921 gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat
1981 gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat
2041 gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg
2101 ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta
2161 agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa
2221 attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc
2281 atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg
2341 gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggacatctcg
2401 gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc
2461 aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag
2521 ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc
2581 ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
2641 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta
2701 gctcactcat taggcaccgg gatctcgacc gatgcccttg agagccttca acccagtcag
2761 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
2821 catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt
2881 tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct
2941 cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat
3001 cgccggcatg gcggccccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct
3061 aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg
3121 aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt
3181 ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat
3241 ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg
3301 ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt
3361 accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat
3421 cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc
3481 atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac
3541 attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga
3601 atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga
3661 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
3721 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
3781 agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca
3841 gagcagattg tactgagagt gcaccatata tgcggtgtga aataccgcac agatgcgtaa
3901 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
3961 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
4021 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
4081 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
4141 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
4201 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
4261 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
4321 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
4381 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
4441 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
4501 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
4561 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
4621 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
4681 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
4741 aaaactcacg ttaagggatt ttggtcatga acaataaaac tgtctgctta cataaacagt
4801 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgctctag gccgcgatta
4861 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
4921 tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa
4981 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
5041 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
5101 ttactcacca ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat
5161 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
5221 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
5281 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
5341 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
5401 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
5461 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
5521 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
5581 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta agaattaatt
5641 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
5701 atttccccga aaagtgccac ctgaaattgt aaacgttaat attttgttaa aattcgcgtt
5761 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
5821 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
5881 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
5941 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
6001 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
6061 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
6121 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
6181 ccattcgcca
DEFINITION pET-40b(+)
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTOAUTHORAD1|Novagen Inc.|
FEATURES Location/Qualifiers
source 1..6190
/organism="pET-40b(+)"
/mol_type="other DNA"
terminator complement(1..120)
/label="T7_terminator"
misc_feature 69..87
/label="T7_Terminal_primer"
CDS complement(147..1115)
/label="ORF frame 3"
/translation="MKKGFMLFTLLAAFSGFAQADDAAIQQTLAKMGIKSSDIQPAPV
AGMKTVLTNSGVLYITDDGKHIIQGPMYDVSGTAPVNVTNKMLLKQLNALEKEMIVYK
APQEKHVITVFTDITCGYCHKLHEQMADYNALGITVRYLAFPRQGLDSDAEKEMKAIW
CAKDKNKAFDDVMAGKSVAPASCDVDIADHYALGVQLGVSGTPAVVLSNGTLVPGYQP
PKEMKEFLDEHQKMTSGKGSTSGSGHHHHHHSAGLVPRGSTAIGMKETAAAKFERQHM
DSPDLGTDDDDKSPGFSSTMAISDPNSSSVDKLAAALEHHHHHHHH*"
misc_feature complement(252..266)
/label="EK"
/translation="MKKGFMLFTLLAAFSGFAQADDAAIQQTLAKMGIKSSDIQPAPV
AGMKTVLTNSGVLYITDDGKHIIQGPMYDVSGTAPVNVTNKMLLKQLNALEKEMIVYK
APQEKHVITVFTDITCGYCHKLHEQMADYNALGITVRYLAFPRQGLDSDAEKEMKAIW
CAKDKNKAFDDVMAGKSVAPASCDVDIADHYALGVQLGVSGTPAVVLSNGTLVPGYQP
PKEMKEFLDEHQKMTSGKGSTSGSGHHHHHHSAGLVPRGSTAIGMKETAAAKFERQHM
DSPDLGTDDDDKSPGFSSTMAISDPNSSSVDKLAAALEHHHHHHHH*"
misc_feature complement(282..326)
/label="S15"
/translation="MKKGFMLFTLLAAFSGFAQADDAAIQQTLAKMGIKSSDIQPAPV
AGMKTVLTNSGVLYITDDGKHIIQGPMYDVSGTAPVNVTNKMLLKQLNALEKEMIVYK
APQEKHVITVFTDITCGYCHKLHEQMADYNALGITVRYLAFPRQGLDSDAEKEMKAIW
CAKDKNKAFDDVMAGKSVAPASCDVDIADHYALGVQLGVSGTPAVVLSNGTLVPGYQP
PKEMKEFLDEHQKMTSGKGSTSGSGHHHHHHSAGLVPRGSTAIGMKETAAAKFERQHM
DSPDLGTDDDDKSPGFSSTMAISDPNSSSVDKLAAALEHHHHHHHH*"
misc_feature complement(369..386)
/label="6xHIS"
/translation="MKKGFMLFTLLAAFSGFAQADDAAIQQTLAKMGIKSSDIQPAPV
AGMKTVLTNSGVLYITDDGKHIIQGPMYDVSGTAPVNVTNKMLLKQLNALEKEMIVYK
APQEKHVITVFTDITCGYCHKLHEQMADYNALGITVRYLAFPRQGLDSDAEKEMKAIW
CAKDKNKAFDDVMAGKSVAPASCDVDIADHYALGVQLGVSGTPAVVLSNGTLVPGYQP
PKEMKEFLDEHQKMTSGKGSTSGSGHHHHHHSAGLVPRGSTAIGMKETAAAKFERQHM
DSPDLGTDDDDKSPGFSSTMAISDPNSSSVDKLAAALEHHHHHHHH*"
CDS 1116..1844
/label="ORF frame 3"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
misc_feature complement(1124..1140)
/label="T7_transl_en_RBS"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
misc_feature complement(1158..1185)
/label="lacO"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
promoter complement(1185..1203)
/label="T7_promoter"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
gene 1239..1502
/label="tet (300 - 563)"
/gene="tet (300 - 563)"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
misc_feature complement(1291..1310)
/label="pBRrevBam_primer"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
misc_feature 1585..2676
/label="lacI"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
CDS 1717..2676
/label="ORF frame 1"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
gene 2735..3010
/label="tet (576 - 851)"
/gene="tet (576 - 851)"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
misc_feature 3485..3676
/label="ROP"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
misc_feature complement(3670..3692)
/label="pGEX_3_primer"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
rep_origin complement(4091..4710)
/label="pBR322_origin"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
gene 4816..5631
/label="KanR2"
/gene="KanR2"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
CDS 4816..5631
/label="ORF frame 1"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgtttagcag cctaggtatt aatcaattag tggtggtggt ggtggtggtg gtgctcgagt
181 gcggccgcaa gcttgtcgac ggagctcgaa ttcggatccg atatcgccat ggttgaggag
241 aagcccgggc tcttgtcgtc gtcatcggta cccagatctg ggctgtccat gtgctggcgt
301 tcgaatttag cagcagcggt ttctttcata ccaattgcag tactaccgcg tggcaccaga
361 cccgcggagt gatggtgatg gtgatgacca gaaccactag ttgatccttt accgctggtc
421 attttttggt gttcgtcgag aaattctttc atctctttcg gcggctggta acccggaaca
481 agtgtgccat tgctcagcac aactgccgga gtaccgctaa cgccaagctg gacgccaagt
541 gcgtaatggt cggcaatatc cacgtcgcaa ctggctggtg cgacgctttt acctgccatc
601 acatcatcaa acgctttgtt tttatctttc gcacaccaga tagctttcat ttctttctct
661 gcatcgctgt ccagcccctg gcgcgggaaa gcaagataac gcacggtgat ccccagcgcg
721 ttgtagtctg ccatttgctc atgcagtttg tggcagtaac cacaggtaat atcagtaaac
781 acggtgatga cgtgtttttc ctgcggcgct ttataaacga tcatctcttt ttcaagcgca
841 ttcaactgct ttaacagcat cttattggtg acattgaccg gagccgtgcc actaacgtca
901 tacattggcc cctgaatgat atgtttacca tcatcggtga tgtacaacac gccgctgtta
961 gtcagaactg tcttcatgcc agctacaggc gcgggctgaa tatcgctgct tttgatgccc
1021 attttggcta acgtttgttg aattgccgcg tcatcagcct gagcaaagcc tgaaaacgcc
1081 gctaacaaag taaacaacat aaaacctttc ttcatatgta tatctccttc ttaaagttaa
1141 acaaaattat ttctagaggg gaattgttat ccgctcacaa ttcccctata gtgagtcgta
1201 ttaatttcgc gggatcgaga tcgatctcga tcctctacgc cggacgcatc gtggccggca
1261 tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag
1321 atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc
1381 ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg
1441 tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag
1501 agcgtcgaga tcccggacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag
1561 cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc
1621 gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac
1681 gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc
1741 aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc
1801 agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa
1861 ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg
1921 gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat
1981 gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat
2041 gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg
2101 ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta
2161 agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa
2221 attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc
2281 atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg
2341 gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggacatctcg
2401 gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc
2461 aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag
2521 ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc
2581 ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
2641 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta
2701 gctcactcat taggcaccgg gatctcgacc gatgcccttg agagccttca acccagtcag
2761 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
2821 catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt
2881 tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct
2941 cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat
3001 cgccggcatg gcggccccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct
3061 aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg
3121 aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt
3181 ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat
3241 ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg
3301 ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt
3361 accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat
3421 cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc
3481 atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac
3541 attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga
3601 atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga
3661 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
3721 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
3781 agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca
3841 gagcagattg tactgagagt gcaccatata tgcggtgtga aataccgcac agatgcgtaa
3901 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
3961 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
4021 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
4081 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
4141 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
4201 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
4261 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
4321 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
4381 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
4441 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
4501 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
4561 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
4621 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
4681 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
4741 aaaactcacg ttaagggatt ttggtcatga acaataaaac tgtctgctta cataaacagt
4801 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgctctag gccgcgatta
4861 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
4921 tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa
4981 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
5041 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
5101 ttactcacca ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat
5161 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
5221 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
5281 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
5341 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
5401 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
5461 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
5521 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
5581 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta agaattaatt
5641 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
5701 atttccccga aaagtgccac ctgaaattgt aaacgttaat attttgttaa aattcgcgtt
5761 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
5821 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
5881 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
5941 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
6001 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
6061 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
6121 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
6181 ccattcgcca
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表达载体的构建是生物技术和所需蛋白质产生的基本工具，本文总结了pET-32α（+）载体技术的构建，该技术通常用于研究实验室。该方法包括获得用于构建载体的外源DNA片段，将DNA片段亚克隆到pET-32α（+）表达载体中，在大肠杆菌 BL21（DE3）中进行蛋白质表达以及在大肠杆菌中在天然条件下进行蛋白质纯化。裂解液。介绍作为分子生物学实验室中非常有用的实践，重组DNA技术（或基因克隆）是指将DNA片段从一种生物体转移到自我复制的遗传元件如表达载体。然后插入的DNA可以在外源宿主细胞中繁殖
相关专题 1)噬菌体DE3溶源化的菌株如BL21(DE3)是最常用的表达菌株，噬菌体DE3是λ噬菌体的衍生株，构建好的表达载体 可以直接转入表达菌株中，诱导蛋白的表达方式与lac启动子一样都是IPTG诱导。 2)表达菌株BL21(DE3)上带有lacI基因，T7 RNA 聚合酶编码基因以及lacUV5启动子，lacUV5启动子可以启动T7 RNA 聚合酶的表达; 3)表达质粒 如pET-28质粒带有T7启动子和编码阻遏
Expression System、HisPatch ThioFusion System、pBAD Inducible Bacterial System、pL System和T7-based Systems。这四个系列的载体可谓把原核表达的发展史中出现过的几种启动子都囊括进去了，包括T7、pL、pBAD和trc启动子，同时几种经典的调控方式也都出现了，包括有乳糖操纵子、色氨酸操纵子和阿拉伯糖操纵子。 冠军系列 Champion™ pET Expression System之所以取
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pET-40b(+)载体
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