LOCUS       Exported                5394 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pCoGWS, complete sequence.
ACCESSION   JX262233
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5394
                     /organism="Expression vector pCoGWS"
                     /mol_type="other DNA"
                     /db_xref="taxon:1230379"
     promoter        7..25
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      7..24
                     /regulatory_class="promoter"
     regulatory      26..50
                     /regulatory_class="attenuator"
     protein_bind    26..50
                     /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      80..86
                     /regulatory_class="ribosome_binding_site"
     CDS             106..123
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    139..263
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /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..1031
                     /codon_start=1
                     /gene="cmR"
                     /product="chloramphenicol resistant protein"
                     /label=cmR
                     /protein_id="AFR34393.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            372..1031
                     /gene="cmR"
                     /label=cmR
     CDS             1373..1678
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB"
                     /label=ccdB
                     /protein_id="AFR34394.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            1373..1678
                     /gene="ccdB"
                     /label=ccdB
     protein_bind    complement(1719..1843)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /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
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      2098..2145
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      2099..2145
                     /regulatory_class="terminator"
     CDS             complement(2316..3107)
                     /codon_start=1
                     /gene="specR"
                     /product="spectinomycin resistant protein"
                     /label=specR
                     /protein_id="AFR34395.1"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            complement(2316..3107)
                     /gene="specR"
                     /label=specR
     promoter        complement(3108..3199)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3247..3985)
                     /direction=LEFT
     CDS             complement(4195..5277)
                     /codon_start=1
                     /gene="lacI"
                     /product="LacI"
                     /label=lacI
                     /protein_id="AFR34396.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(4195..5277)
                     /gene="lacI"
                     /label=lacI
     promoter        complement(5278..5355)
                     /gene="lacI"
                     /label=lacI promoter