| 出品公司: | EMD Biosciences (Novagen) |
|---|---|
| 别名: | pET29c, pet 29c |
| 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: | 5372 bp |
| 5' 测序引物: | T7 |
| 5' 测序引物序列: | 5'-TAATACGACTCACTATAGGG-3' |
| 载体标签: | N-S, N-Thrombin, C-His |
| 载体抗性: | Kanamycin |
| 表达宿主菌: | BL21(DE3) 、BLR(DE3)、BL21(DE3)pLysS |
| 备注: | N term thrombin cleavage site; a,b,c vary by MCS; |
| 产品目录号: | ZY69873-3 |
| 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-29c(+)
- 库存：
20
- 供应商：
泽叶生物
pET29c载体基本信息
pET29c载体质粒图谱和多克隆位点信息
| Feature Name | Start | End | |
|---|---|---|---|
| T7_terminator | 129 | 1 | |
| T7_Terminal_primer | 69 | 87 | |
| lacI | 219 | 1858 | |
| T7_transl_en_RBS | 322 | 306 | |
| lacO | 367 | 340 | |
| T7_promoter | 385 | 367 | |
| tet (300 - 563) | 421 | 684 | |
| pBRrevBam_primer | 492 | 473 | |
| tet (576 - 851) | 1917 | 2192 | |
| ROP | 2667 | 2858 | |
| pGEX_3_primer | 2874 | 2852 | |
| pBR322_origin | 3892 | 3273 | |
| KanR2 | 3998 | 4813 |
| ORF | Start | End | |
|---|---|---|---|
| ORF frame 1 | 298 | 1026 | |
| ORF frame 2 | 899 | 1858 | |
| ORF frame 2 | 3998 | 4813 |
| Enzyme Name | Cut |
|---|---|
| XhoI | 158 |
| NotI | 166 |
| EagI | 166 |
| HindIII | 173 |
| SalI | 179 |
| SacI | 190 |
| EcoRI | 192 |
| BamHI | 198 |
| EcoRV | 209 |
| NcoI | 213 |
| KpnI | 239 |
| BglII | 242 |
| BstBI | 269 |
| NdeI | 296 |
| XbaI | 334 |
| BclI | 1140 |
| ApaI | 1337 |
| HpaI | 1632 |
| FspI | 2208 |
| NruI | 4086 |
| SmaI | 4303 |
| XmaI | 4301 |
pET29c载体简介
The pET-29a-c(+) vectors carry an N-terminal S•Tag™/thrombin configuration plus an optional C-terminal His•Tag® sequence. 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 circular 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).
pET29c载体序列
LOCUS pET-29 c (+) 5372 bp DNA Circular SYN
DEFINITION pET-29 c (+)
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..5372
/organism="pET-29 c (+)"
/mol_type="other DNA"
terminator complement(1..129)
/label="T7_terminator"
misc_feature 69..87
/label="T7_Terminal_primer"
misc_feature 219..1858
/label="lacI"
CDS 298..1026
/label="ORF frame 1"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
misc_feature complement(306..322)
/label="T7_transl_en_RBS"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
misc_feature complement(340..367)
/label="lacO"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
promoter complement(367..385)
/label="T7_promoter"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
gene 421..684
/label="tet (300 - 563)"
/gene="tet (300 - 563)"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
misc_feature complement(473..492)
/label="pBRrevBam_primer"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
CDS 899..1858
/label="ORF frame 2"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
gene 1917..2192
/label="tet (576 - 851)"
/gene="tet (576 - 851)"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
misc_feature 2667..2858
/label="ROP"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
misc_feature complement(2852..2874)
/label="pGEX_3_primer"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
rep_origin complement(3273..3892)
/label="pBR322_origin"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
gene 3998..4813
/label="KanR2"
/gene="KanR2"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
CDS 3998..4813
/label="ORF frame 2"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
181 cgacggagct cgaattcgga tccacagata tcccatggaa ccgcgtggca ccagggtacc
241 cagatctggg ctgtccatgt gctggcgttc gaatttagca gcagcggttt ctttcatatg
301 tatatctcct tcttaaagtt aaacaaaatt atttctagag gggaattgtt atccgctcac
361 aattccccta tagtgagtcg tattaatttc gcgggatcga gatcgatctc gatcctctac
421 gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc
481 gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc
541 ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat
601 gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta
661 atgcaggagt cgcataaggg agagcgtcga gatcccggac accatcgaat ggcgcaaaac
721 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa
781 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg
841 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat
901 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt
961 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc
1021 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag
1081 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct
1141 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
1201 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
1261 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
1321 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
1381 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
1441 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
1501 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
1561 gcgcgttggt gcggacatct cggtagtggg atacgacgat accgaagaca gctcatgtta
1621 tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
1681 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
1741 actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt
1801 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
1861 gcaacgcaat taatgtaagt tagctcactc attaggcacc gggatctcga ccgatgccct
1921 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg
1981 cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg
2041 tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg
2101 tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt
2161 tcggcgagaa gcaggccatt atcgccggca tggcggcccc acgggtgcgc atgatcgtgc
2221 tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct tactggttag cagaatgaat
2281 caccgatacg cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa
2341 caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc
2401 cctgcaccat tatgttccgg atctgcatcg caggatgctg ctggctaccc tgtggaacac
2461 ctacatctgt attaacgaag cgctggcatt gaccctgagt gatttttctc tggtcccgcc
2521 gcatccatac cgccagttgt ttaccctcac aacgttccag taaccgggca tgttcatcat
2581 cagtaacccg tatcgtgagc atcctctctc gtttcatcgg tatcattacc cccatgaaca
2641 gaaatccccc ttacacggag gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg
2701 cccgctttat cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg
2761 atgaacaggc agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct
2821 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
2881 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
2941 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata
3001 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tatgcggtgt
3061 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
3121 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
3181 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
3241 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
3301 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
3361 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
3421 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
3481 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
3541 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
3601 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
3661 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
3721 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
3781 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
3841 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
3901 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gaacaataaa
3961 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
4021 gtcttgctct aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
4081 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga
4141 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
4201 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
4261 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccgggaaaa cagcattcca
4321 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
4381 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
4441 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
4501 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
4561 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
4621 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
4681 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
4741 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
4801 tgagtttttc taagaattaa ttcatgagcg gatacatatt tgaatgtatt tagaaaaata
4861 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgaaatt gtaaacgtta
4921 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
4981 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
5041 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
5101 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
5161 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
5221 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
5281 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
5341 atgcgccgct acagggcgcg tcccattcgc ca
DEFINITION pET-29 c (+)
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..5372
/organism="pET-29 c (+)"
/mol_type="other DNA"
terminator complement(1..129)
/label="T7_terminator"
misc_feature 69..87
/label="T7_Terminal_primer"
misc_feature 219..1858
/label="lacI"
CDS 298..1026
/label="ORF frame 1"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
misc_feature complement(306..322)
/label="T7_transl_en_RBS"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
misc_feature complement(340..367)
/label="lacO"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
promoter complement(367..385)
/label="T7_promoter"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
gene 421..684
/label="tet (300 - 563)"
/gene="tet (300 - 563)"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
misc_feature complement(473..492)
/label="pBRrevBam_primer"
/translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
TVVADWRCHLQSGPARAVANCRGD*"
CDS 899..1858
/label="ORF frame 2"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
gene 1917..2192
/label="tet (576 - 851)"
/gene="tet (576 - 851)"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
misc_feature 2667..2858
/label="ROP"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
misc_feature complement(2852..2874)
/label="pGEX_3_primer"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
rep_origin complement(3273..3892)
/label="pBR322_origin"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
gene 3998..4813
/label="KanR2"
/gene="KanR2"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
CDS 3998..4813
/label="ORF frame 2"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
181 cgacggagct cgaattcgga tccacagata tcccatggaa ccgcgtggca ccagggtacc
241 cagatctggg ctgtccatgt gctggcgttc gaatttagca gcagcggttt ctttcatatg
301 tatatctcct tcttaaagtt aaacaaaatt atttctagag gggaattgtt atccgctcac
361 aattccccta tagtgagtcg tattaatttc gcgggatcga gatcgatctc gatcctctac
421 gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc
481 gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc
541 ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat
601 gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta
661 atgcaggagt cgcataaggg agagcgtcga gatcccggac accatcgaat ggcgcaaaac
721 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa
781 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg
841 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat
901 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt
961 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc
1021 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag
1081 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct
1141 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
1201 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
1261 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
1321 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
1381 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
1441 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
1501 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
1561 gcgcgttggt gcggacatct cggtagtggg atacgacgat accgaagaca gctcatgtta
1621 tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
1681 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
1741 actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt
1801 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
1861 gcaacgcaat taatgtaagt tagctcactc attaggcacc gggatctcga ccgatgccct
1921 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg
1981 cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg
2041 tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg
2101 tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt
2161 tcggcgagaa gcaggccatt atcgccggca tggcggcccc acgggtgcgc atgatcgtgc
2221 tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct tactggttag cagaatgaat
2281 caccgatacg cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa
2341 caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc
2401 cctgcaccat tatgttccgg atctgcatcg caggatgctg ctggctaccc tgtggaacac
2461 ctacatctgt attaacgaag cgctggcatt gaccctgagt gatttttctc tggtcccgcc
2521 gcatccatac cgccagttgt ttaccctcac aacgttccag taaccgggca tgttcatcat
2581 cagtaacccg tatcgtgagc atcctctctc gtttcatcgg tatcattacc cccatgaaca
2641 gaaatccccc ttacacggag gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg
2701 cccgctttat cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg
2761 atgaacaggc agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct
2821 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
2881 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
2941 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata
3001 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tatgcggtgt
3061 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
3121 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
3181 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
3241 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
3301 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
3361 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
3421 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
3481 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
3541 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
3601 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
3661 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
3721 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
3781 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
3841 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
3901 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gaacaataaa
3961 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
4021 gtcttgctct aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
4081 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga
4141 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
4201 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
4261 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccgggaaaa cagcattcca
4321 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
4381 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
4441 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
4501 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
4561 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
4621 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
4681 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
4741 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
4801 tgagtttttc taagaattaa ttcatgagcg gatacatatt tgaatgtatt tagaaaaata
4861 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgaaatt gtaaacgtta
4921 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
4981 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
5041 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
5101 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
5161 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
5221 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
5281 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
5341 atgcgccgct acagggcgcg tcccattcgc ca
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文献和实验相关实验
加热灭活酶。通过凝胶提取试剂盒纯化消化的pET-32α（+）载体骨架和从pMD18-T载体切下的基因片段。用T4 DNA连接酶在23℃下用pET-32α（+）载体骨架连接基因片段1小时，载体末端和插入末端的比例为1：3（fmol）。将10μl连接反应与100μl感受态大肠杆菌 BL21（DE3）在冰上混合10分钟，并将混合物在42℃下孵育90秒。加入800μlLB并在37°C下孵育40分钟。涂在含有100μg/ ml氨苄青霉素的LB平板上。在37°C孵育过夜。通过菌落PCR筛选插入物的存在或制备
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
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
技术资料暂无技术资料 索取技术资料
pET-29c(+)载体
¥800
