LOCUS       40924_12600        7033 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pCoGWA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7033)
  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  JOURNAL   Unpublished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 7033)
  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 7033)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7033)
  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            "Unpublished"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..7033
                     /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)"
     CDS             106..123
                     /label=6xHis
                     /note="6xHis affinity tag"
     protein_bind    139..263
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        288..318
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             372..1028
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             1373..1675
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(1719..1843)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1921..1939
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1921..1938
                     /regulatory_class="promoter"
     regulatory      1940..1964
                     /regulatory_class="attenuator"
     protein_bind    1940..1964
                     /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      1992..1998
                     /regulatory_class="ribosome_binding_site"
     CDS             2034..2057
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     terminator      2098..2145
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2182..2637
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(2755..3612)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3613..3705)
                     /label=AmpR promoter
     rep_origin      3786..4374
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4560..4702)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4807..4995)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(5533..5554)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(5570..6649)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(6650..6727)
                     /label=lacI promoter
ORIGIN
        1 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa
       61 taattttgtt taactttaag aaggagatat accatgggca gcagccatca ccaccaccac
      121 cacagcggta cctacatcac aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat
      181 ataaatatca atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa
      241 acacaacata tccagtcact atggcggccg cattaggcac cccaggcttt acactttatg
      301 cttccggctc gtataatgtg tggattttga gttaggatcc gtcgagattt tcaggagcta
      361 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
      421 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
      481 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
      541 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa
      601 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
      661 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
      721 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
      781 ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg
      841 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt
      901 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
      961 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
     1021 gcggggcgta aacgcgtgga tccggcttac taaaagccag ataacagtat gcgtatttgc
     1081 gcgctgattt ttgcggtata agaatatata ctgatatgta tacccgaagt atgtcaaaaa
     1141 gaggtatgct atgaagcagc gtattacagt gacagttgac agcgacagct atcagttgct
     1201 caaggcatat atgatgtcaa tatctccggt ctggtaagca caaccatgca gaatgaagcc
     1261 cgtcgtctgc gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc
     1321 cggtttattg aaatgaacgg ctcttttgct gacgagaaca ggggctggtg aaatgcagtt
     1381 taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga
     1441 tattattgac acgcccgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc
     1501 agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat
     1561 gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag tggctgatct
     1621 cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat
     1681 gtcaggctcc cttatacaca gccagtctgc aggtcgacca tagtgactgg atatgttgtg
     1741 ttttacagta ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta
     1801 tatcatttta cgtttctcgt tcagctttct tgtacaaagt ggtgatgtaa agcttgcggc
     1861 cgcataatgc ttaagtcgaa cagaaagtaa tcgtattgta cacggccgca taatcgaaat
     1921 taatacgact cactataggg gaattgtgag cggataacaa ttccccatct tagtatatta
     1981 gttaagtata agaaggagat atacatatgg aattccctag gagatctctc gaggactaca
     2041 aggacgacga tgacaaataa ctcgagggtc taggctgctg ccaccgctga gcaataacta
     2101 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
     2161 atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2221 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     2281 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2341 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     2401 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     2461 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     2521 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     2581 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttctg
     2641 gcggcacgat ggcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2701 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2761 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     2821 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     2881 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     2941 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3001 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3061 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3121 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3181 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3241 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3301 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3361 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3421 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3481 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3541 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     3601 ttgaatactc atactcttcc tttttcaatc atgattgaag catttatcag ggttattgtc
     3661 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggt catgaccaaa
     3721 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3781 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3841 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3901 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     3961 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4021 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4081 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4141 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4201 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4261 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4321 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4381 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     4441 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     4501 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     4561 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atatggtgca
     4621 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     4681 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     4741 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     4801 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     4861 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     4921 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     4981 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
     5041 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     5101 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     5161 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
     5221 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
     5281 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
     5341 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
     5401 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgctagtcat
     5461 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
     5521 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     5581 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5641 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
     5701 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
     5761 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
     5821 cggtatcgtc gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa
     5881 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
     5941 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
     6001 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
     6061 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
     6121 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
     6181 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
     6241 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
     6301 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
     6361 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
     6421 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
     6481 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
     6541 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
     6601 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     6661 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
     6721 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt
     6781 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg
     6841 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     6901 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     6961 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc
     7021 gatctcgatc ccg
//
