pCOLADuet-1

pCOLADuet-1

pCOLADuet1载体基本信息

出品公司:	Ybscience
别名:	pCOLADuet1, pCOLADuet 1
质粒类型:	大肠杆菌蛋白双表达载体，有两个多克隆位点
表达水平:	高
克隆方法:	多克隆位点，限制性内切酶
载体大小:	3719 bp
5' 测序引物:	T7
5' 测序引物序列:	T7: 5'-TAATACGACTCACTATAGGG-3';
3' 测序引物:	T7t
3' 测序引物序列:	T7t: 5'-TGCTAGTTATTGCTCAGCGG-3'
载体标签:	N-His, C-S
载体抗性:	Kanamycin
备注:	pRSFDuet-1 carries the RSF1030 replicon, lacI gene and kanamycin resistance gene.
稳定性:	瞬时表达 Transient
组成型:	组成型 Constitutive
病毒/非病毒:	非病毒

pCOLADuet1载体质粒图谱和多克隆位点信息

pCOLADuet1载体简介

pCOLADuet-1 is designed for the coexpression of two target genes from a single plasmid. The vector encodes two multiple cloning sites (MCS) each of which is preceded by a T7 promoter, lac operator, and ribosome binding site (rbs). MCS-1 encodes the six-amino acid His•Tag sequence for the creation of a N-terminal fusion and MCS2 encodes the 15 amino acid S•Tag™ peptide after the last restriction site for the creation of a C-terminal fusion if desired. Genes inserted into MCS-1 can be sequenced using the ACYCDuetUP1 Primer and DuetDOWN1 Primer. Genes inserted into MCS-2 can be sequenced using the DuetUP2 Primer and T7 Terminator Primer. The vector has the COLA replicon from ColA(1) and the kanamycin resistance gene. This vector can be transformed into the same cell with plasmids containing compatible origins of replication and drug resistance genes for coexpression of up to 8 target genes.

pCOLADuet1载体序列

LOCUS       pCOLADuet-1    3719 bp     DNA  Circular   SYN
DEFINITION  pCOLADuet-1
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..3719
                     /organism="pCOLADuet-1"
                     /mol_type="other DNA"
     misc_feature    2..29
                     /label="lacO"
     promoter        214..232
                     /label="T7_promoter"
     misc_feature    232..259
                     /label="lacO"
     misc_feature    complement(448..466)
                     /label="T7_Terminal_primer"
     gene            complement(739..1554)
                     /label="KanR2"
                     /gene="KanR2"
     CDS             complement(739..1554)
                     /label="ORF frame 3"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
                     GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
                     GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
                     VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
                     RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
     promoter        complement(1596..1624)
                     /label="AmpR_promoter"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
                     GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
                     GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
                     VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
                     RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
     misc_feature    complement(2497..3588)
                     /label="lacI"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
                     GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
                     GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
                     VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
                     RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
     CDS             complement(2497..3456)
                     /label="ORF frame 3"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     CDS             complement(3329..158)
                     /label="ORF frame 1"
                     /translation="MRPQACRPAGAPSSNSDPGCGDDGDGCCPWYISLLKLNKIISTG
                     ELLSAHNSPIVSRINFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECET
                     SNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTV
                     VADWRCHLQSGPARAVANCRGD*"
     promoter        3703..2
                     /label="T7_promoter"
                     /translation="MRPQACRPAGAPSSNSDPGCGDDGDGCCPWYISLLKLNKIISTG
                     ELLSAHNSPIVSRINFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECET
                     SNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTV
                     VADWRCHLQSGPARAVANCRGD*"
ORIGIN
    1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
   61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag
  121 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag
  181 taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt
  241 gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga gatatacata
  301 tggcagatct caattggata tcggccggcc acgcgatcgc tgacgtcggt accctcgagt
  361 ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg
  421 cagcttaatt aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg
  481 cctctaaacg ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg
  541 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt
  601 aacgaccctg ccctgaaccg acgacaagct gacgaccggg tctccgcaag tggcactttt
  661 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
  721 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
  781 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
  841 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
  901 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
  961 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
 1021 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
 1081 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cggtcgctgt taaaaggaca
 1141 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
 1201 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
 1261 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
 1321 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
 1381 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
 1441 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
 1501 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcatactctt
 1561 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
 1621 tgaatgtatt tagaaaaata aacaaatagg catgctagcg cagaaacgtc ctagaagatg
 1681 ccaggaggat acttagcaga gagacaataa ggccggagcg aagccgtttt tccataggct
 1741 ccgcccccct gacgaacatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac
 1801 aggactataa agataccagg cgtttccccc tgatggctcc ctcttgcgct ctcctgttcc
 1861 cgtcctgcgg cgtccgtgtt gtggtggagg ctttacccaa atcaccacgt cccgttccgt
 1921 gtagacagtt cgctccaagc tgggctgtgt gcaagaaccc cccgttcagc ccgactgctg
 1981 cgccttatcc ggtaactatc atcttgagtc caacccggaa agacacgaca aaacgccact
 2041 ggcagcagcc attggtaact gagaattagt ggatttagat atcgagagtc ttgaagtggt
 2101 ggcctaacag aggctacact gaaaggacag tatttggtat ctgcgctcca ctaaagccag
 2161 ttaccaggtt aagcagttcc ccaactgact taaccttcga tcaaaccgcc tccccaggcg
 2221 gttttttcgt ttacagagca ggagattacg acgatcgtaa aaggatctca agaagatcct
 2281 ttacggattc ccgacaccat cactctagat ttcagtgcaa tttatctctt caaatgtagc
 2341 acctgaagtc agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc
 2401 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
 2461 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
 2521 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
 2581 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
 2641 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
 2701 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
 2761 gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
 2821 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
 2881 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
 2941 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
 3001 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
 3061 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
 3121 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
 3181 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
 3241 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
 3301 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
 3361 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
 3421 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
 3481 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
 3541 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
 3601 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
 3661 gatctcgacg ctctccctta tgcgactcct gcattaggaa attaatacga ctcactata