LOCUS       40924_2343        7105 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector p0GWA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7105)
  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 7105)
  AUTHORS   Busso D, Delagoutte-Busso B, Salim L, Moras D.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-APR-2008) Structural Biology and Genomics Platform, 
            IGBMC, 1, rue Laurent Fries, Illkirch 67404, France
REFERENCE   3  (bases 1 to 7105)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7105)
  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 
            (29-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..7105
                     /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)"
     protein_bind    135..259
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        284..314
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             368..1024
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1369..1671
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(1715..1839)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1853..1870
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      1937..1984
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2021..2476
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2503..2607
                     /label=AmpR promoter
     CDS             2608..3465
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3639..4227
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4413..4555)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4660..4848)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(5623..5644)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(5660..6739)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(6740..6817)
                     /label=lacI promoter
ORIGIN
        1 agatccagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
       61 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat accatgggta
      121 ccggatctta catcacaagt ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa
      181 atatcaatat attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac
      241 aacatatcca gtcactatgg cggccgcatt aggcacccca ggctttacac tttatgcttc
      301 cggctcgtat aatgtgtgga ttttgagtta ggatccgtcg agattttcag gagctaagga
      361 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
      421 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
      481 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
      541 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa
      601 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
      661 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
      721 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
      781 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
      841 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
      901 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg
      961 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
     1021 ggcgtaaacg cgtggatccg gcttactaaa agccagataa cagtatgcgt atttgcgcgc
     1081 tgatttttgc ggtataagaa tatatactga tatgtatacc cgaagtatgt caaaaagagg
     1141 tatgctatga agcagcgtat tacagtgaca gttgacagcg acagctatca gttgctcaag
     1201 gcatatatga tgtcaatatc tccggtctgg taagcacaac catgcagaat gaagcccgtc
     1261 gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg gatggctgag gtcgcccggt
     1321 ttattgaaat gaacggctct tttgctgacg agaacagggg ctggtgaaat gcagtttaag
     1381 gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt
     1441 attgacacgc ccgggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat
     1501 aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg
     1561 accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc
     1621 caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca
     1681 ggctccctta tacacagcca gtctgcaggt cgaccatagt gactggatat gttgtgtttt
     1741 acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc
     1801 attttacgtt tctcgttcag ctttcttgta caaagtggtg atgtacctcg agcaccacca
     1861 ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     1921 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     1981 tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca
     2041 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2101 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2161 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2221 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2281 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2341 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2401 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     2461 ttaacgttta caatttcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     2521 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2581 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     2641 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     2701 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     2761 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     2821 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     2881 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     2941 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3001 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3061 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3121 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     3181 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     3241 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3301 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     3361 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     3421 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     3481 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     3541 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     3601 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3661 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3721 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3781 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3841 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3901 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3961 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4021 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4081 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4141 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4201 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4261 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4321 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     4381 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     4441 ctgtgcggta tttcacaccg catatatggt gcactctcag tacaatctgc tctgatgccg
     4501 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     4561 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     4621 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     4681 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     4741 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     4801 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     4861 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     4921 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     4981 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     5041 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
     5101 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
     5161 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     5221 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     5281 cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc
     5341 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
     5401 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
     5461 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
     5521 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     5581 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     5641 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5701 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     5761 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     5821 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     5881 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat
     5941 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     6001 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     6061 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     6121 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     6181 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     6241 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     6301 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     6361 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     6421 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     6481 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     6541 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     6601 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     6661 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     6721 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     6781 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
     6841 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
     6901 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     6961 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     7021 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     7081 acgatgcgtc cggcgtagag gatcg
//
