LOCUS       40924_47173        7435 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pXGWA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7435)
  AUTHORS   Busso D, Delagoutte-Busso B, Moras D.
  TITLE     Construction of a set Gateway-based destination vectors for 
            high-throughput cloning and expression screening in Escherichia coli
  JOURNAL   Anal. Biochem. 343 (2), 313-321 (2005)
  PUBMED    15993367
REFERENCE   2  (bases 1 to 7435)
  AUTHORS   Busso D, Delagoutte-Busso B, Salim L, Moras D.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-APR-2008) Structural Biology and Genomics Platform, 
            IGBMC, 1, rue Laurent Fries, Illkirch 67404, France
REFERENCE   3  (bases 1 to 7435)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7435)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Anal. 
            Biochem."; date: "2005"; volume: "343"; issue: "2"; pages: "313-321"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-APR-2008) Structural Biology and Genomics Platform, IGBMC, 1, 
            rue Laurent Fries, Illkirch 67404, France"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..45
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    46..70
                     /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             85..107
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             123..446
                     /codon_start=1
                     /label=TrxA
                     /note="E. coli thioredoxin"
                     /translation="SDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDEI
                     ADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFLD
                     ANLA"
     protein_bind    465..589
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        614..644
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             698..1354
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1699..2001
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2045..2169)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2183..2200
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      2267..2314
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2351..2806
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2833..2937
                     /label=AmpR promoter
     CDS             2938..3795
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3969..4557
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4743..4885)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4990..5178)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(5953..5974)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(5990..7069)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(7070..7147)
                     /label=lacI promoter
ORIGIN
        1 agatccagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
       61 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat accatgggta
      121 ccagcgataa aattattcac ctgactgacg acagttttga cacggatgta ctcaaagcgg
      181 acggggcgat cctcgtcgat ttctgggcag agtggtgcgg tccgtgcaaa atgatcgccc
      241 cgattctgga tgaaatcgct gacgaatatc agggcaaact gaccgttgca aaactgaaca
      301 tcgatcaaaa ccctggcact gcgccgaaat atggcatccg tggtatcccg actctgctgc
      361 tgttcaaaaa cggtgaagtg gcggcaacca aagtgggtgc actgtctaaa ggtcagttga
      421 aagagttcct cgacgctaac ctggccggta ccggatctta catcacaagt ttgtacaaaa
      481 aagctgaacg agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata
      541 aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgg cggccgcatt
      601 aggcacccca ggctttacac tttatgcttc cggctcgtat aatgtgtgga ttttgagtta
      661 ggatccgtcg agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata
      721 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
      781 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
      841 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
      901 tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
      961 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
     1021 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
     1081 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
     1141 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
     1201 ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
     1261 gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
     1321 acaacagtac tgcgatgagt ggcagggcgg ggcgtaaacg cgtggatccg gcttactaaa
     1381 agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa tatatactga
     1441 tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat tacagtgaca
     1501 gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc tccggtctgg
     1561 taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa
     1621 atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct tttgctgacg
     1681 agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga gagccgttat
     1741 cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg gatggtgatc
     1801 cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg
     1861 catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc
     1921 gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt
     1981 aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca gtctgcaggt
     2041 cgaccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa
     2101 aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag ctttcttgta
     2161 caaagtggtg atgtacctcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa
     2221 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     2281 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt
     2341 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2401 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2461 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2521 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2581 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2641 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2701 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2761 caaaaattta acgcgaattt taacaaaata ttaacgttta caatttcagg tggcactttt
     2821 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2881 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     2941 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3001 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3061 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3121 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3181 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3241 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3301 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3361 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     3421 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     3481 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3541 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3601 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3661 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3721 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3781 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     3841 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3901 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3961 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4021 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4081 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     4141 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4201 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4261 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4321 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4381 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4441 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4501 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4561 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4621 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     4681 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     4741 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatatggt
     4801 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     4861 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     4921 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4981 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     5041 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     5101 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     5161 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat
     5221 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
     5281 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
     5341 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag
     5401 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt
     5461 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga
     5521 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc
     5581 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac
     5641 ccgtggggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
     5701 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
     5761 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
     5821 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
     5881 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
     5941 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     6001 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6061 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
     6121 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
     6181 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
     6241 cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa
     6301 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
     6361 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
     6421 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
     6481 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
     6541 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
     6601 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
     6661 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
     6721 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
     6781 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
     6841 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
     6901 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
     6961 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
     7021 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     7081 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
     7141 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt
     7201 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg
     7261 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     7321 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     7381 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcg
//
