LOCUS       40924_12610        5394 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pCoGWS, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5394)
  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  JOURNAL   Unpublished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 5394)
  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 5394)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5394)
  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            "Unpublished"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..5394
                     /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(2319..3107)
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     promoter        complement(3108..3199)
                     /label=AmpR promoter
     rep_origin      complement(3247..3985)
                     /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(4161..4182)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4198..5277)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(5278..5355)
                     /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 ccgggtcatc gtggccggat
     2281 cttgcggccc ctcggcttga acgaattgtt agacattatt tgccgactac cttggtgatc
     2341 tcgcctttca cgtagtggac aaattcttcc aactgatctg cgcgcgaggc caagcgatct
     2401 tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata ctgggccggc
     2461 aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc ggttactgcg
     2521 ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc ccagtcgggc
     2581 ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg ttcaggaacc
     2641 ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc tcttgctttt
     2701 gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc tgcaagaatg
     2761 tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg ccacggaatg
     2821 atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct ctctccaggg
     2881 gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt
     2941 acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc atccactgcg
     3001 gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat gacgccaact
     3061 acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcatact cttccttttt
     3121 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3181 atttagaaaa ataaacaaat agctagctca ctcggtcgct acgctccggg cgtgagactg
     3241 cggcgggcgc tgcggacaca tacaaagtta cccacagatt ccgtggataa gcaggggact
     3301 aacatgtgag gcaaaacagc agggccgcgc cggtggcgtt tttccatagg ctccgccctc
     3361 ctgccagagt tcacataaac agacgctttt ccggtgcatc tgtgggagcc gtgaggctca
     3421 accatgaatc tgacagtacg ggcgaaaccc gacaggactt aaagatcccc accgtttccg
     3481 gcgggtcgct ccctcttgcg ctctcctgtt ccgaccctgc cgtttaccgg atacctgttc
     3541 cgcctttctc ccttacggga agtgtggcgc tttctcatag ctcacacact ggtatctcgg
     3601 ctcggtgtag gtcgttcgct ccaagctggg ctgtaagcaa gaactccccg ttcagcccga
     3661 ctgctgcgcc ttatccggta actgttcact tgagtccaac ccggaaaagc acggtaaaac
     3721 gccactggca gcagccattg gtaactggga gttcgcagag gatttgttta gctaaacacg
     3781 cggttgctct tgaagtgtgc gccaaagtcc ggctacactg gaaggacaga tttggttgct
     3841 gtgctctgcg aaagccagtt accacggtta agcagttccc caactgactt aaccttcgat
     3901 caaaccacct ccccaggtgg ttttttcgtt tacagggcaa aagattacgc gcagaaaaaa
     3961 aggatctcaa gaagatcctt tgatcttttc tactgaaccg ctctagattt cagtgcaatt
     4021 tatctcttca aatgtagcac ctgaagtcag ccccatacga tataagttgt aattctcatg
     4081 ttagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     4141 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     4201 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4261 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     4321 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     4381 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     4441 gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc gcagcccgga
     4501 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     4561 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     4621 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     4681 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     4741 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     4801 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     4861 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     4921 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     4981 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     5041 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     5101 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     5161 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     5221 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     5281 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
     5341 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc atta
//
