PortEco logo

pCOLADuet-1

From EcoliWiki
Jump to: navigation, search

At a Glance

pCOLADuet-1, Image made with PlasMapper

Type

Coexpression

Origin of Replication

ColA

Copy #
Markers

Kanamycin

Link to Sequence

Novagen

Features

T7 Promoter

His Tag

S Tag

2 MCS

lacI

RBS

Accession Number




Notes

pCOLADuet™-1 is designed for the coexpression of two target genes from a single plasmid. The vector has the COLA replicon from ColA[1]. 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 eight target genes[2][3]. Plasmids that share common origins of replication or antibiotic resistance genes should not be used for the coexpression of two proteins[4].

History

pCOLADuet-1 for coexpression was created in order to provide a useful alternative to the complicated and expensive expression systems, such as yeast, baculovirus or mammalian cell culture, which are commonly used to overcome problems of heterologous protein expression.

Sources

Novagen

References

[back to top]


See Help:References for how to manage references in EcoliWiki.

  1. Zverev, VV & Khmel', IA (1985) [Nucleotide sequence of the replicative region of the plasmid ColA-CA31]. Mol. Gen. Mikrobiol. Virusol. 18-24 PubMed EcoliWiki page
  2. Novy, R., et all. 2002. inNovations 15,2.
  3. Held, D., et al. 2003. inNovations 18, 4.
  4. Tolia, NH & Joshua-Tor, L (2006) Strategies for protein coexpression in Escherichia coli. Nat. Methods 3 55-64 PubMed EcoliWiki page

Sequence

   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