LOCUS       40924_25201        7462 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pHXGWA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7462)
  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 7462)
  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 7462)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7462)
  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..7462
                     /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             126..143
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             150..473
                     /codon_start=1
                     /label=TrxA
                     /note="E. coli thioredoxin"
                     /translation="SDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDEI
                     ADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFLD
                     ANLA"
     protein_bind    492..616
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        641..671
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             725..1381
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1726..2028
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2072..2196)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2210..2227
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      2294..2341
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2378..2833
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2860..2964
                     /label=AmpR promoter
     CDS             2965..3822
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3996..4584
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4770..4912)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(5017..5205)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(5980..6001)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(6017..7096)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(7097..7174)
                     /label=lacI promoter
ORIGIN
        1 agatccagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
       61 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat accatgggca
      121 gcagccatca tcatcatcat cacggtacca gcgataaaat tattcacctg actgacgaca
      181 gttttgacac ggatgtactc aaagcggacg gggcgatcct cgtcgatttc tgggcagagt
      241 ggtgcggtcc gtgcaaaatg atcgccccga ttctggatga aatcgctgac gaatatcagg
      301 gcaaactgac cgttgcaaaa ctgaacatcg atcaaaaccc tggcactgcg ccgaaatatg
      361 gcatccgtgg tatcccgact ctgctgctgt tcaaaaacgg tgaagtggcg gcaaccaaag
      421 tgggtgcact gtctaaaggt cagttgaaag agttcctcga cgctaacctg gccggtaccg
      481 gatcttacat cacaagtttg tacaaaaaag ctgaacgaga aacgtaaaat gatataaata
      541 tcaatatatt aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac
      601 atatccagtc actatggcgg ccgcattagg caccccaggc tttacacttt atgcttccgg
      661 ctcgtataat gtgtggattt tgagttagga tccgtcgaga ttttcaggag ctaaggaagc
      721 taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa
      781 agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct
      841 ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt
      901 tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga
      961 cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac
     1021 tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat
     1081 atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat
     1141 tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa
     1201 cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca
     1261 aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt
     1321 ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc
     1381 gtaaacgcgt ggatccggct tactaaaagc cagataacag tatgcgtatt tgcgcgctga
     1441 tttttgcggt ataagaatat atactgatat gtatacccga agtatgtcaa aaagaggtat
     1501 gctatgaagc agcgtattac agtgacagtt gacagcgaca gctatcagtt gctcaaggca
     1561 tatatgatgt caatatctcc ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc
     1621 tgcgtgccga acgctggaaa gcggaaaatc aggaagggat ggctgaggtc gcccggttta
     1681 ttgaaatgaa cggctctttt gctgacgaga acaggggctg gtgaaatgca gtttaaggtt
     1741 tacacctata aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt
     1801 gacacgcccg ggcgacggat ggtgatcccc ctggccagtg cacgtctgct gtcagataaa
     1861 gtctcccgtg aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc
     1921 accgatatgg ccagtgtgcc ggtctccgtt atcggggaag aagtggctga tctcagccac
     1981 cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc
     2041 tcccttatac acagccagtc tgcaggtcga ccatagtgac tggatatgtt gtgttttaca
     2101 gtattatgta gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt
     2161 ttacgtttct cgttcagctt tcttgtacaa agtggtgatg tacctcgagc accaccacca
     2221 ccaccactga gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     2281 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     2341 gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta
     2401 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2461 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2521 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2581 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2641 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2701 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2761 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2821 acgtttacaa tttcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2881 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     2941 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3001 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3061 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3121 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     3181 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     3241 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3301 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3361 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3421 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3481 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     3541 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     3601 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     3661 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     3721 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     3781 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     3841 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3901 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3961 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4021 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4081 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4141 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4201 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4261 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4321 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4381 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4441 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4501 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4561 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4621 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4681 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     4741 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     4801 tgcggtattt cacaccgcat atatggtgca ctctcagtac aatctgctct gatgccgcat
     4861 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
     4921 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     4981 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     5041 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     5101 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     5161 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta
     5221 agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat
     5281 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
     5341 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa
     5401 tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat
     5461 aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac
     5521 cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc
     5581 gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct
     5641 caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg
     5701 cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa
     5761 gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc
     5821 gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac
     5881 agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     5941 gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt
     6001 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     6061 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     6121 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     6181 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     6241 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg
     6301 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     6361 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     6421 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     6481 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     6541 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     6601 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     6661 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     6721 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     6781 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     6841 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     6901 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     6961 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     7021 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     7081 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     7141 taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc
     7201 gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca
     7261 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     7321 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     7381 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
     7441 atgcgtccgg cgtagaggat cg
//
