LOCUS       40924_21848        7768 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pGGWA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7768)
  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 7768)
  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 7768)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7768)
  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..7768
                     /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..773
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="SPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKF
                     ELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYG
                     VSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLY
                     MDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     protein_bind    798..922
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        947..977
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1031..1687
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2032..2334
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2378..2502)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2516..2533
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      2600..2647
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2684..3139
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3166..3270
                     /label=AmpR promoter
     CDS             3271..4128
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4302..4890
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(5076..5218)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(5323..5511)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(6286..6307)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(6323..7402)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(7403..7480)
                     /label=lacI promoter
ORIGIN
        1 agatccagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
       61 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat accatgggta
      121 cctcccctat actaggttat tggaaaatta agggccttgt gcaacccact cgacttcttt
      181 tggaatatct tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa ggtgataaat
      241 ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa tcttccttat tatattgatg
      301 gtgatgttaa attaacacag tctatggcca tcatacgtta tatagctgac aagcacaaca
      361 tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga gcggttttgg
      421 atattagata cggtgtttcg agaattgcat atagtaaaga ctttgaaact ctcaaagttg
      481 attttcttag caagctacct gaaatgctga aaatgttcga agatcgttta tgtcataaaa
      541 catatttaaa tggtgatcat gtaacccatc ctgacttcat gttgtatgac gctcttgatg
      601 ttgttttata catggaccca atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa
      661 aacgtattga agctatccca caaattgata agtacttgaa atccagcaag tatatagcat
      721 ggcctttgca gggctggcaa gccacgtttg gtggtggcga ccatccccca aaatcggatg
      781 gtaccggatc ttacatcaca agtttgtaca aaaaagctga acgagaaacg taaaatgata
      841 taaatatcaa tatattaaat tagattttgc ataaaaaaca gactacataa tactgtaaaa
      901 cacaacatat ccagtcacta tggcggccgc attaggcacc ccaggcttta cactttatgc
      961 ttccggctcg tataatgtgt ggattttgag ttaggatccg tcgagatttt caggagctaa
     1021 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca
     1081 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt
     1141 tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
     1201 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc gtatggcaat
     1261 gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga
     1321 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct
     1381 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg
     1441 gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga
     1501 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta
     1561 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     1621 tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg
     1681 cggggcgtaa acgcgtggat ccggcttact aaaagccaga taacagtatg cgtatttgcg
     1741 cgctgatttt tgcggtataa gaatatatac tgatatgtat acccgaagta tgtcaaaaag
     1801 aggtatgcta tgaagcagcg tattacagtg acagttgaca gcgacagcta tcagttgctc
     1861 aaggcatata tgatgtcaat atctccggtc tggtaagcac aaccatgcag aatgaagccc
     1921 gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga agggatggct gaggtcgccc
     1981 ggtttattga aatgaacggc tcttttgctg acgagaacag gggctggtga aatgcagttt
     2041 aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt acagagtgat
     2101 attattgaca cgcccgggcg acggatggtg atccccctgg ccagtgcacg tctgctgtca
     2161 gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg
     2221 atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt ggctgatctc
     2281 agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg aatataaatg
     2341 tcaggctccc ttatacacag ccagtctgca ggtcgaccat agtgactgga tatgttgtgt
     2401 tttacagtat tatgtagtct gttttttatg caaaatctaa tttaatatat tgatatttat
     2461 atcattttac gtttctcgtt cagctttctt gtacaaagtg gtgatgtacc tcgagcacca
     2521 ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     2581 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     2641 ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc
     2701 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2761 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2821 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2881 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2941 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3001 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3061 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     3121 atattaacgt ttacaatttc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3181 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3241 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3301 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3361 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3421 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3481 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3541 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3601 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3661 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3721 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3781 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     3841 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3901 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3961 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4021 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4081 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4141 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4201 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4261 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4321 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4381 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4441 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4501 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4561 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4621 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4681 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4741 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4801 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4861 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4921 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4981 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5041 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     5101 catctgtgcg gtatttcaca ccgcatatat ggtgcactct cagtacaatc tgctctgatg
     5161 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     5221 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     5281 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     5341 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     5401 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     5461 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     5521 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     5581 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     5641 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     5701 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
     5761 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     5821 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     5881 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     5941 ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc cggcgataat
     6001 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     6061 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     6121 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     6181 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     6241 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     6301 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6361 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     6421 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     6481 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     6541 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     6601 tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     6661 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     6721 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     6781 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     6841 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     6901 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     6961 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     7021 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     7081 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     7141 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     7201 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     7261 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     7321 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     7381 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     7441 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
     7501 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
     7561 gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct
     7621 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
     7681 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
     7741 gccacgatgc gtccggcgta gaggatcg
//
