LOCUS       40924_12605        5621 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pCoGWC, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5621)
  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  JOURNAL   Unpublished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 5621)
  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 5621)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5621)
  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            "Unpublished"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..5621
                     /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"
     CDS             complement(2368..3024)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(3025..3127)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      complement(3653..4197)
                     /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(4388..4409)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4425..5504)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(5505..5582)
                     /label=lacI promoter
ORIGIN
        1 ggaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctgtagaaa
       61 taattttgtt taactttaat 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 acctcaggca
     2161 tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag
     2221 acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa tttgctttcg
     2281 aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag gcgtttaagg
     2341 gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc agtactgttg
     2401 taattcatta agcattctgc cgacatggaa gccatcacag acggcatgat gaacctgaat
     2461 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatag tgaaaacggg
     2521 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg
     2581 attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc
     2641 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta
     2701 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg
     2761 aacactatcc catatcacca gctcaccgtc tttcattgcc atacggaact ccggatgagc
     2821 attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt
     2881 tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc
     2941 aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt
     3001 atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc
     3061 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     3121 ccgatcaacg tctcattttc gccaaaagtt ggcccagggc ttcccggtat caacagggac
     3181 accaggattt atttattctg cgaagtgatc ttccgtcaca ggtatttatt cggcgcaaag
     3241 tgcgtcgggt gatgctgcca acttactgat ttagtgtatg atggtgtttt tgaggtgctc
     3301 cagtggcttc tgtttctatc agctgtccct cctgttcagc tactgacggg gtggtgcgta
     3361 acggcaaaag caccgccgga catcagcgct agcggagtgt atactggctt actatgttgg
     3421 cactgatgag ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg
     3481 cgtcagcaga atatgtgata caggatatat tccgcttcct cgctcactga ctcgctacgc
     3541 tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga tttcctggaa
     3601 gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg tttttccata
     3661 ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc
     3721 cgacaggact ataaagatac caggcgtttc ccctggcggc tccctcgtgc gctctcctgt
     3781 tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac
     3841 gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc
     3901 ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa
     3961 gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga gttagtcttg
     4021 aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca
     4081 agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc
     4141 aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa
     4201 gatcatctta ttaatcagat aaaatatttc tagatttcag tgcaatttat ctcttcaaat
     4261 gtagcacctg aagtcagccc catacgatat aagttgtaat tctcatgtta gtcatgcccc
     4321 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     4381 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     4441 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4501 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     4561 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     4621 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     4681 tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc ggtaatggcg
     4741 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     4801 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     4861 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     4921 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     4981 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     5041 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     5101 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     5161 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     5221 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     5281 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     5341 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     5401 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     5461 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     5521 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     5581 tccgggatct cgacgctctc ccttatgcga ctcctgcatt a
//
