| 出品公司: | -- |
|---|---|
| 载体名称: | pMCSG7 |
| 质粒类型: | 细菌表达 |
| 高拷贝/低拷贝: | -- |
| 复制子: | pBR322 |
| 启动子: | T7 |
| 载体大小: | 5286 bp |
| 5' 测序引物及序列: | -- |
| 3' 测序引物及序列: | -- |
| 载体标签: | -- |
| 载体抗性: | 氨苄青霉素 |
| 筛选标记: | -- |
| 克隆菌株: | DH5α 等 |
| 宿主细胞（系）: | -- |
| 备注: | -- |
| 产品目录号: | -- |
| 稳定性: | -- |
| 组成型/诱导型: | -- |
| 病毒/非病毒: | -- |
买家导航
pMCSG7载体质粒基本信息
pMCSG7质粒图谱载体图谱和pMCSG7载体序列质粒序列多克隆位点信息
pMCSG7质粒载体简介
pMCSG7质粒序列载体序列
LOCUS pMCSG7 5286 bp ds-DNA circular SYN 27-JAN-2014
DEFINITION Bacterial vector with a 6xHis-TEV leader for high-throughput
purification of recombinant proteins.
ACCESSION .
VERSION .
KEYWORDS pMCSG7
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5286)
AUTHORS Stols L, Gu M, Dieckman L, Raffen R, Collart FR, Donnelly MI.
TITLE A new vector for high-throughput, ligation-independent cloning
encoding a tobacco etch virus protease cleavage site.
JOURNAL Protein Expr. Purif. 2002;25:8-15.
PUBMED 12071693
REFERENCE 2 (bases 1 to 5286)
AUTHORS Midwest Center for Structural Genomics
TITLE Direct Submission
JOURNAL Exported Friday, February 28, 2020 from SnapGene Viewer 3.1.4
http://www.snapgene.com
COMMENT For ligation-independent cloning (LIC), linearize with SspI and
treat with T4 DNA polymerase plus dGTP.
FEATURES Location/Qualifiers
source 1..5286
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
terminator 26..73
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(140..157)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(223..243)
/codon_start=1
/product="tobacco etch virus (TEV) recognition and cleavage
site"
/note="TEV site"
/translation="ENLYFQS"
CDS complement(268..285)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(286..288)
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
RBS 297..302
/note="ribosome binding site"
protein_bind 333..357
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(358..376)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 689..766
/gene="lacI"
/note="lacI promoter"
CDS 767..1849
/codon_start=1
/gene="lacI"
/product="lac repressor"
/note="lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
CDS 2658..2849
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
rep_origin complement(3279..3867)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4038..4898)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4899..5002)
/gene="bla"
/note="AmpR promoter"
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 tccgttatcc acttccaata ttggattgga agtacaggtt
241 ctcggtaccc agatctacac cagaagaatg atgatgatga tggtgcatat gtatatctcc
301 ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca caattcccct
361 atagtgagtc gtattaattt cgcgggatcg agatcgatct cgatcctcta cgccggacgc
421 atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc
481 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt
541 atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc
601 cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag
661 tcgcataagg gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa cctttcgcgg
721 tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac
781 gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa
841 ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct
901 gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg
961 cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc
1021 tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga
1081 agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa
1141 ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc
1201 gttatttctt gatgtctctg accagacacc catcaacagt attattttct cccatgaaga
1261 cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt
1321 agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct
1381 cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg
1441 ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc
1501 caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg
1561 tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc
1621 gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct
1681 gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa
1741 aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
1801 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
1861 ttaatgtaag ttagctcact cattaggcac cgggatctcg accgatgccc ttgagagcct
1921 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
1981 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg
2041 gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa
2101 tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga
2161 agcaggccat tatcgccggc atggcggccc cacgggtgcg catgatcgtg ctcctgtcgt
2221 tgaggacccg gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac
2281 gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa
2341 tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca
2401 ttatgttccg gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg
2461 tattaacgaa gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata
2521 ccgccagttg tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc
2581 gtatcgtgag catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc
2641 cttacacgga ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta
2701 tcagaagcca gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg
2761 cagacatctg tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc
2821 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
2881 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
2941 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa
3001 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atatgcggtg tgaaataccg
3061 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
3121 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
3181 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
3241 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
3301 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
3361 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
3421 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
3481 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
3541 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
3601 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
3661 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
3721 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
3781 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
3841 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
3901 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
3961 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
4021 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
4081 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
4141 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
4201 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
4261 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
4321 gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg
4381 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
4441 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
4501 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
4561 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
4621 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
4681 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
4741 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
4801 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
4861 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaaattatt
4921 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
4981 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt
5041 taatgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
5101 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
5161 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
5221 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
5281 tcgcca
//
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