LOCUS       40924_12580        7012 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pCo0GWA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7012)
  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  JOURNAL   Unpublished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 7012)
  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 7012)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7012)
  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            "Unpublished"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..7012
                     /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"
     rep_origin      2161..2616
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(2734..3591)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3592..3684)
                     /label=AmpR promoter
     rep_origin      3765..4353
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4539..4681)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4786..4974)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(5512..5533)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(5549..6628)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(6629..6706)
                     /label=lacI promoter
ORIGIN
        1 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa
       61 taattttgtt taactttaag 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 ggaggaacta tatccggatt ggcgaatggg
     2161 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2221 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2281 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2341 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2401 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2461 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     2521 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     2581 acgcgaattt taacaaaata ttaacgttta caatttctgg cggcacgatg gcatgagatt
     2641 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2701 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2761 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2821 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2881 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2941 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3001 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3061 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3121 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3181 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3241 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3301 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3361 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3421 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3481 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3541 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3601 ttttcaatca tgattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3661 aatgtattta gaaaaataaa caaataggtc atgaccaaaa tcccttaacg tgagttttcg
     3721 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3781 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3841 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3901 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3961 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4021 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4081 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4141 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4201 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4261 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4321 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4381 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4441 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4501 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     4561 acgcatctgt gcggtatttc acaccgcata tatggtgcac tctcagtaca atctgctctg
     4621 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     4681 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4741 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4801 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     4861 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     4921 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     4981 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     5041 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     5101 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg
     5161 cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat
     5221 ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa
     5281 accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca
     5341 cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag
     5401 ccgggtcctc aacgacagga gcacgatcat gctagtcatg ccccgcgccc accggaagga
     5461 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
     5521 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5581 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     5641 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
     5701 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
     5761 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
     5821 ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
     5881 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     5941 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     6001 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
     6061 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
     6121 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
     6181 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     6241 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     6301 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     6361 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     6421 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     6481 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     6541 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     6601 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     6661 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc
     6721 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
     6781 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     6841 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     6901 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     6961 atgccggcca cgatgcgtcc ggcgtagagg atcgagatcg atctcgatcc cg
//
