LOCUS       40924_22769        5037 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pGST-parallel1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5037)
  AUTHORS   Sheffield P, Garrard S, Derewenda Z.
  TITLE     Overcoming expression and purification problems of RhoGDI using a 
            family of 'parallel' expression vectors
  JOURNAL   Protein Expr. Purif. 15 (1), 34-39 (1999)
  PUBMED    10024467
REFERENCE   2  (bases 1 to 5037)
  AUTHORS   Sheffield PJ, Garrard SM, Derewenda ZS.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-OCT-1998) Molecular Physiology and Biological Physics,
            University of Virginia, 4215 Jordan Hall, 1300 Jefferson Park 
            Avenue, Charlottesville, Virginia 22908, USA
REFERENCE   3  (bases 1 to 5037)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5037)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein 
            Expr. Purif."; date: "1999"; volume: "15"; issue: "1"; pages: 
            "34-39"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (05-OCT-1998) Molecular Physiology and Biological Physics, 
            University of Virginia, 4215 Jordan Hall, 1300 Jefferson Park 
            Avenue, Charlottesville, Virginia 22908, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        183..211
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    219..235
                     /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)."
     CDS             258..911
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             924..944
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     misc_feature    946..1034
                     /label=multiple cloning site
                     /note="multiple cloning site"
     promoter        1340..1444
                     /label=AmpR promoter
     CDS             1445..2302
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2476..3064
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        3308..3385
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             3386..4465
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    4481..4502
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4517..4547
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4555..4571
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     CDS             4591..4764
                     /codon_start=1
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
ORIGIN
        1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa aatcgatacg accgaaaacc tgtattttca gggcgccatg gatccggaat
      961 tcaaaggcct acgtcgacga gctcaactag tgcggccgct ttcgaatcta gagcctgcag
     1021 tctcgagcgg ccgcatcgtg actgactgac gatctgcctc gcgcgtttcg gtgatgacgg
     1081 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     1141 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
     1201 catgacccag tcacgtagcg atagcggagt gtataattct tgaagacgaa agggcctcgt
     1261 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     1321 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     1381 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     1441 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     1501 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     1561 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     1621 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     1681 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     1741 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     1801 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     1861 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     1921 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1981 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     2041 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     2101 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     2161 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     2221 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     2281 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     2341 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     2401 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2461 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2521 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2581 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     2641 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2701 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2761 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2821 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2881 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2941 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3001 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3061 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3121 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3181 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     3241 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3301 aaattccgac accatcgaat ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga
     3361 agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta
     3421 tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc
     3481 gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt
     3541 ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc
     3601 cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc
     3661 cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca
     3721 caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga
     3781 tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga
     3841 ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga
     3901 gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt
     3961 ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc
     4021 gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat
     4081 gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg
     4141 cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg
     4201 atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga
     4261 ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc
     4321 ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc
     4381 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     4441 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     4501 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     4561 gcggataaca atttcacaca ggaaacagct atgaccatga ttacggattc actggccgtc
     4621 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     4681 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     4741 cagttgcgca gcctgaatgg cgaatggcgc tttgcctggt ttccggcacc agaagcggtg
     4801 ccggaaagct ggctggagtg cgatcttcct gaggccgata ctgtcgtcgt cccctcaaac
     4861 tggcagatgc acggttacga tgcgcccatc tacaccaacg taacctatcc cattacggtc
     4921 aatccgccgt ttgttcccac ggagaatccg acgggttgtt actcgctcac atttaatgtt
     4981 gatgaaagct ggctacagga aggccagacg cgaattattt ttgatggcgt tggaatt
//
