LOCUS       40924_12585        5600 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pCo0GWC, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5600)
  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  JOURNAL   Unpublished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 5600)
  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 5600)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5600)
  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            "Unpublished"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..5600
                     /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(2347..3003)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(3004..3106)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      complement(3632..4176)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     protein_bind    complement(4367..4388)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4404..5483)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(5484..5561)
                     /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 cgggtcgaat ttgctttcga atttctgcca ttcatccgct
     2281 tattatcact tattcaggcg tagcaccagg cgtttaaggg caccaataac tgccttaaaa
     2341 aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc
     2401 gacatggaag ccatcacaga cggcatgatg aacctgaatc gccagcggca tcagcacctt
     2461 gtcgccttgc gtataatatt tgcccatagt gaaaacgggg gcgaagaagt tgtccatatt
     2521 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat
     2581 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
     2641 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
     2701 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag
     2761 ctcaccgtct ttcattgcca tacggaactc cggatgagca ttcatcaggc gggcaagaat
     2821 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt
     2881 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa
     2941 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc
     3001 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     3061 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt ctcattttcg
     3121 ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta tttattctgc
     3181 gaagtgatct tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa
     3241 cttactgatt tagtgtatga tggtgttttt gaggtgctcc agtggcttct gtttctatca
     3301 gctgtccctc ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc accgccggac
     3361 atcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg gtgtcagtga
     3421 agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac
     3481 aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga
     3541 gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac
     3601 agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc
     3661 atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc
     3721 aggcgtttcc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg
     3781 tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt
     3841 aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg
     3901 ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg
     3961 cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc
     4021 taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa
     4081 gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca
     4141 gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata
     4201 aaatatttct agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc
     4261 atacgatata agttgtaatt ctcatgttag tcatgccccg cgcccaccgg aaggagctga
     4321 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
     4381 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4441 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg
     4501 gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga
     4561 gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg
     4621 gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag
     4681 atgtccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc
     4741 tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt
     4801 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
     4861 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
     4921 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
     4981 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
     5041 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
     5101 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
     5161 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
     5221 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
     5281 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
     5341 atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg
     5401 ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca
     5461 tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
     5521 catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc
     5581 cttatgcgac tcctgcatta
//
