LOCUS       40924_12590        5373 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pCo0GWS, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5373)
  AUTHORS   Busso D.
  TITLE     Construction of a set of Gateway-based co-expression vectors for 
            rapid cloning and multi-protein production in Escherichia coli
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5373)
  AUTHORS   Busso D.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JUN-2012) Molecular Biology, IGBMC, 1 rue Laurent 
            Fries, Illkirch, Alsace 67404, France
REFERENCE   3  (bases 1 to 5373)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5373)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-JUN-2012) Molecular Biology, IGBMC, 1 rue Laurent Fries, 
            Illkirch, Alsace 67404, France"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..25
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    26..50
                     /label=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)."
     RBS             65..87
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    118..242
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        267..297
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             351..1007
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             1352..1654
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(1698..1822)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1900..1918
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1900..1917
                     /regulatory_class="promoter"
     regulatory      1919..1943
                     /regulatory_class="attenuator"
     protein_bind    1919..1943
                     /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)."
     regulatory      1971..1977
                     /regulatory_class="ribosome_binding_site"
     CDS             2013..2036
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     terminator      2077..2124
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2298..3086)
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     promoter        complement(3087..3178)
                     /label=AmpR promoter
     rep_origin      complement(3226..3964)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     protein_bind    complement(4140..4161)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4177..5256)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(5257..5334)
                     /label=lacI promoter
ORIGIN
        1 ggaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctgtagaaa
       61 taattttgtt taactttaat aaggagatat accatgggca gcagcggtac ctacatcaca
      121 agtttgtaca aaaaagctga acgagaaacg taaaatgata taaatatcaa tatattaaat
      181 tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat ccagtcacta
      241 tggcggccgc attaggcacc ccaggcttta cactttatgc ttccggctcg tataatgtgt
      301 ggattttgag ttaggatccg tcgagatttt caggagctaa ggaagctaaa atggagaaaa
      361 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg
      421 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct
      481 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg
      541 cccgcctgat gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga
      601 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat
      661 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg
      721 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt
      781 tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg
      841 acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc
      901 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
      961 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa acgcgtggat
     1021 ccggcttact aaaagccaga taacagtatg cgtatttgcg cgctgatttt tgcggtataa
     1081 gaatatatac tgatatgtat acccgaagta tgtcaaaaag aggtatgcta tgaagcagcg
     1141 tattacagtg acagttgaca gcgacagcta tcagttgctc aaggcatata tgatgtcaat
     1201 atctccggtc tggtaagcac aaccatgcag aatgaagccc gtcgtctgcg tgccgaacgc
     1261 tggaaagcgg aaaatcagga agggatggct gaggtcgccc ggtttattga aatgaacggc
     1321 tcttttgctg acgagaacag gggctggtga aatgcagttt aaggtttaca cctataaaag
     1381 agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcccgggcg
     1441 acggatggtg atccccctgg ccagtgcacg tctgctgtca gataaagtct cccgtgaact
     1501 ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag
     1561 tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat
     1621 caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggctccc ttatacacag
     1681 ccagtctgca ggtcgaccat agtgactgga tatgttgtgt tttacagtat tatgtagtct
     1741 gttttttatg caaaatctaa tttaatatat tgatatttat atcattttac gtttctcgtt
     1801 cagctttctt gtacaaagtg gtgatgtaaa gcttgcggcc gcataatgct taagtcgaac
     1861 agaaagtaat cgtattgtac acggccgcat aatcgaaatt aatacgactc actatagggg
     1921 aattgtgagc ggataacaat tccccatctt agtatattag ttaagtataa gaaggagata
     1981 tacatatgga attccctagg agatctctcg aggactacaa ggacgacgat gacaaataac
     2041 tcgagggtct aggctgctgc caccgctgag caataactag cataacccct tggggcctct
     2101 aaacgggtct tgaggggttt tttgctgaaa cctcaggcat ttgagaagca cacggtcaca
     2161 ctgcttccgg tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga
     2221 ccctgccctg aaccgacgac cgggtcatcg tggccggatc ttgcggcccc tcggcttgaa
     2281 cgaattgtta gacattattt gccgactacc ttggtgatct cgcctttcac gtagtggaca
     2341 aattcttcca actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc
     2401 tgtctagctt caagtatgac gggctgatac tgggccggca ggcgctccat tgcccagtcg
     2461 gcagcgacat ccttcggcgc gattttgccg gttactgcgc tgtaccaaat gcgggacaac
     2521 gtaagcacta catttcgctc atcgccagcc cagtcgggcg gcgagttcca tagcgttaag
     2581 gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc
     2641 gctggaccta ccaaggcaac gctatgttct cttgcttttg tcagcaagat agccagatca
     2701 atgtcgatcg tggctggctc gaagatacct gcaagaatgt cattgcgctg ccattctcca
     2761 aattgcagtt cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg cacaacaatg
     2821 gtgacttcta cagcgcggag aatctcgctc tctccagggg aagccgaagt ttccaaaagg
     2881 tcgttgatca aagctcgccg cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa
     2941 tcaatatcac tgtgtggctt caggccgcca tccactgcgg agccgtacaa atgtacggcc
     3001 agcaacgtcg gttcgagatg gcgctcgatg acgccaacta cctctgatag ttgagtcgat
     3061 acttcggcga tcaccgcttc cctcatactc ttcctttttc aatattattg aagcatttat
     3121 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     3181 gctagctcac tcggtcgcta cgctccgggc gtgagactgc ggcgggcgct gcggacacat
     3241 acaaagttac ccacagattc cgtggataag caggggacta acatgtgagg caaaacagca
     3301 gggccgcgcc ggtggcgttt ttccataggc tccgccctcc tgccagagtt cacataaaca
     3361 gacgcttttc cggtgcatct gtgggagccg tgaggctcaa ccatgaatct gacagtacgg
     3421 gcgaaacccg acaggactta aagatcccca ccgtttccgg cgggtcgctc cctcttgcgc
     3481 tctcctgttc cgaccctgcc gtttaccgga tacctgttcc gcctttctcc cttacgggaa
     3541 gtgtggcgct ttctcatagc tcacacactg gtatctcggc tcggtgtagg tcgttcgctc
     3601 caagctgggc tgtaagcaag aactccccgt tcagcccgac tgctgcgcct tatccggtaa
     3661 ctgttcactt gagtccaacc cggaaaagca cggtaaaacg ccactggcag cagccattgg
     3721 taactgggag ttcgcagagg atttgtttag ctaaacacgc ggttgctctt gaagtgtgcg
     3781 ccaaagtccg gctacactgg aaggacagat ttggttgctg tgctctgcga aagccagtta
     3841 ccacggttaa gcagttcccc aactgactta accttcgatc aaaccacctc cccaggtggt
     3901 tttttcgttt acagggcaaa agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3961 gatcttttct actgaaccgc tctagatttc agtgcaattt atctcttcaa atgtagcacc
     4021 tgaagtcagc cccatacgat ataagttgta attctcatgt tagtcatgcc ccgcgcccac
     4081 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
     4141 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4201 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4261 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
     4321 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     4381 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
     4441 tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
     4501 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     4561 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     4621 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     4681 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     4741 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
     4801 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     4861 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     4921 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     4981 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
     5041 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     5101 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     5161 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     5221 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
     5281 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat
     5341 ctcgacgctc tcccttatgc gactcctgca tta
//
