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pMSCG19载体质粒基本信息
pMSCG19质粒图谱载体图谱和pMSCG19载体序列质粒序列多克隆位点信息
pMSCG19质粒载体简介
pMSCG19质粒序列载体序列
LOCUS pMSCG19 6441 bp DNA circular SYN 07-MAR-2024
DEFINITION Bacterial vector with an MBP-TVMV-6xHis-TEV leader for
high-throughput purification of recombinant proteins.
ACCESSION .
VERSION .
KEYWORDS pMCSG19
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6441)
AUTHORS Donnelly MI, Zhou M, Millard CS, Clancy S, Stols L, Eschenfeldt WH,
Collart FR, Joachimiak A.
TITLE An expression vector tailored for large-scale, high-throughput
purification of recombinant proteins.
JOURNAL Protein Expr. Purif. 2006;47:446-54.
PUBMED 16497515
REFERENCE 2 (bases 1 to 6441)
AUTHORS Midwest Center for Structural Genomics
TITLE Direct Submission
JOURNAL Exported May 8, 2024 from SnapGene 6.0.2
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Protein
Expr. Purif."; date: "2006"; volume: "47"; pages: "446-54"
COMMENT For ligation-independent cloning (LIC), linearize with SspI and
treat with T4 DNA polymerase plus dGTP.
FEATURES Location/Qualifiers
source 1..6441
/lab_host="Escherichia coli"
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 26..73
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(140..157)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS complement(223..243)
/codon_start=1
/product="tobacco etch virus (TEV) recognition and cleavage
site"
/label=TEV site
/translation="ENLYFQS"
CDS complement(268..285)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS complement(286..306)
/codon_start=1
/product="tobacco vein mottling virus (TVMV) NIa protease
recognition and cleavage site
"
/label=TVMV site
/translation="ETVRFQS"
CDS complement(343..1443)
/codon_start=1
/gene="malE (mutated)"
/product="maltose binding protein from E. coli"
/label=MBP
/note="This version of the gene does not encode a signal
sequence, so MBP will remain in the cytosol."
/translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
WYAVRTAVINAASGRQTVDEALKDAQT"
RBS 1452..1457
/note="ribosome binding site"
protein_bind 1488..1512
/label=lac operator
/bound_moiety="lac repressor encoded by 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)."
promoter complement(1513..1531)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 1844..1921
/gene="lacI"
/label=lacI promoter
CDS 1922..3004
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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 3813..4004
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
rep_origin complement(4434..5022)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5193..6053)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(6054..6157)
/gene="bla"
/label=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 agatccacgc cagaagagtg atgatgatgg tggtgagact ggaaacgcac
301 ggtttccgag cctgcttttt tgtacaaact tgtgatcgaa ttagtctgcg cgtctttcag
361 ggcttcatcg acagtctgac gaccgctggc ggcgttgatc accgcagtac gcacggcata
421 ccagaaagcg gacatctgcg ggatgttcgg catgatttca cctttctggg cgttttccat
481 ggtggcggca atacgtggat ctttcgccaa ctcttcctcg taagacttca gcgctacggc
541 acccagcggt ttgtctttat taaccgcttc cagaccttca tcagtcagca gatagttttc
601 gaggaactct tttgccagct ctttgttcgg actggcggcg ttaatacctg cgctcagcac
661 gccaacgaac ggtttggatg gttgaccctt gaaggtcggc agtaccgtta caccataatt
721 cactttgctg gtgtcgatgt tggaccatgc ccacgggccg ttgatggtca tcgctgtttc
781 gcctttatta aaggcagctt ctgcgatgga gtaatcggtg tctgcattca tgtgtttgtt
841 tttaatcagg tcaaccagga aggtcagacc cgctttcgcg ccagcgttat ccacgcccac
901 gtctttaatg tcgtacttgc cgttttcata cttgaacgca taacccccgt cagcagcaat
961 cagcggccag gtgaagtacg gttcttgcag gttgaacatc agcgcgctct tacctttcgc
1021 tttcagttct ttatccagcg ccgggatctc ttcccaggtt tttggcgggt tcggcagcag
1081 atctttgtta taaatcagcg ataacgcttc aacagcgatc gggtaagcaa tcagcttgcc
1141 gttgtaacgt acggcatccc aggtaaacgg atacagcttg tcctggaacg ctttgtccgg
1201 ggtgatttca gccaacaggc cagattgagc gtagccacca aagcggtcgt gtgcccagaa
1261 gataatgtca gggccatcgc cagttgccgc aacctgtggg aatttctctt ccagtttatc
1321 cggatgctca acggtgactt taattccggt atctttctcg aatttcttac cgacttcagc
1381 gagaccgtta tagcctttat cgccgttaat ccagattacc agtttacctt cttcgatttt
1441 catatgtata tctccttctt aaagttaaac aaaattattt ctagagggga attgttatcc
1501 gctcacaatt cccctatagt gagtcgtatt aatttcgcgg gatcgagatc gatctcgatc
1561 ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc
1621 tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct
1681 tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc
1741 ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc
1801 ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca tcgaatggcg
1861 caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa
1921 tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt
1981 ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc
2041 ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca
2101 gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt
2161 cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga
2221 acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag
2281 tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg
2341 cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat
2401 tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca
2461 gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg
2521 ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg
2581 gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac
2641 tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc
2701 cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc
2761 atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag
2821 cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc
2881 cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg
2941 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
3001 gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga tctcgaccga
3061 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
3121 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
3181 tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc
3241 ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca
3301 aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccccacgg gtgcgcatga
3361 tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga
3421 atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct
3481 gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt
3541 cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg
3601 gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt
3661 cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt
3721 catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca
3781 tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta
3841 acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg
3901 acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc
3961 gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
4021 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
4081 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
4141 tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatatg
4201 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
4261 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
4321 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
4381 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
4441 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
4501 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
4561 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
4621 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
4681 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
4741 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
4801 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
4861 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
4921 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
4981 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
5041 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
5101 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
5161 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
5221 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
5281 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
5341 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
5401 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
5461 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg
5521 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
5581 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
5641 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
5701 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
5761 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
5821 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
5881 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
5941 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
6001 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
6061 ctttttcaaa ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
6121 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
6181 ctaaattgta agcgttaatg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
6241 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
6301 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
6361 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
6421 cagggcgcgt cccattcgcc a
//
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