LOCUS       40924_24538        7795 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pHGGWA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7795)
  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 7795)
  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 7795)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7795)
  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..7795
                     /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..800
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="SPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKF
                     ELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYG
                     VSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLY
                     MDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     protein_bind    825..949
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        974..1004
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1058..1714
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2059..2361
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2405..2529)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2543..2560
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      2627..2674
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2711..3166
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3193..3297
                     /label=AmpR promoter
     CDS             3298..4155
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4329..4917
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(5103..5245)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(5350..5538)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(6313..6334)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(6350..7429)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(7430..7507)
                     /label=lacI promoter
ORIGIN
        1 agatccagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
       61 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat accatgggca
      121 gcagccatca tcatcatcat cacggtacct cccctatact aggttattgg aaaattaagg
      181 gccttgtgca acccactcga cttcttttgg aatatcttga agaaaaatat gaagagcatt
      241 tgtatgagcg cgatgaaggt gataaatggc gaaacaaaaa gtttgaattg ggtttggagt
      301 ttcccaatct tccttattat attgatggtg atgttaaatt aacacagtct atggccatca
      361 tacgttatat agctgacaag cacaacatgt tgggtggttg tccaaaagag cgtgcagaga
      421 tttcaatgct tgaaggagcg gttttggata ttagatacgg tgtttcgaga attgcatata
      481 gtaaagactt tgaaactctc aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa
      541 tgttcgaaga tcgtttatgt cataaaacat atttaaatgg tgatcatgta acccatcctg
      601 acttcatgtt gtatgacgct cttgatgttg ttttatacat ggacccaatg tgcctggatg
      661 cgttcccaaa attagtttgt tttaaaaaac gtattgaagc tatcccacaa attgataagt
      721 acttgaaatc cagcaagtat atagcatggc ctttgcaggg ctggcaagcc acgtttggtg
      781 gtggcgacca tcccccaaaa tcggatggta ccggatctta catcacaagt ttgtacaaaa
      841 aagctgaacg agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata
      901 aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgg cggccgcatt
      961 aggcacccca ggctttacac tttatgcttc cggctcgtat aatgtgtgga ttttgagtta
     1021 ggatccgtcg agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata
     1081 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
     1141 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
     1201 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
     1261 tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
     1321 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
     1381 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
     1441 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
     1501 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
     1561 ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
     1621 gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
     1681 acaacagtac tgcgatgagt ggcagggcgg ggcgtaaacg cgtggatccg gcttactaaa
     1741 agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa tatatactga
     1801 tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat tacagtgaca
     1861 gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc tccggtctgg
     1921 taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa
     1981 atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct tttgctgacg
     2041 agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga gagccgttat
     2101 cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg gatggtgatc
     2161 cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg
     2221 catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc
     2281 gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt
     2341 aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca gtctgcaggt
     2401 cgaccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa
     2461 aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag ctttcttgta
     2521 caaagtggtg atgtacctcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa
     2581 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     2641 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt
     2701 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2761 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2821 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2881 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2941 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3001 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3061 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3121 caaaaattta acgcgaattt taacaaaata ttaacgttta caatttcagg tggcactttt
     3181 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3241 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3301 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3361 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3421 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3481 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3541 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3601 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3661 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3721 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     3781 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     3841 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3901 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3961 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4021 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4081 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4141 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4201 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4261 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4321 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4381 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4441 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     4501 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4561 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4621 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4681 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4741 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4801 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4861 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4921 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4981 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     5041 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     5101 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatatggt
     5161 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     5221 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     5281 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     5341 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     5401 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     5461 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     5521 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat
     5581 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
     5641 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
     5701 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag
     5761 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt
     5821 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga
     5881 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc
     5941 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac
     6001 ccgtggggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
     6061 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
     6121 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
     6181 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
     6241 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
     6301 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     6361 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6421 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
     6481 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
     6541 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
     6601 cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa
     6661 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
     6721 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
     6781 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
     6841 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
     6901 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
     6961 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
     7021 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
     7081 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
     7141 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
     7201 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
     7261 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
     7321 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
     7381 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     7441 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
     7501 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt
     7561 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg
     7621 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     7681 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     7741 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcg
//
