LOCUS       40924_46133        8008 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pVTLU263, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8008)
  AUTHORS   Mok CSL., Melcher K.
  TITLE     Leu2d-based yeast expression vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8008)
  AUTHORS   Mok CSL., Melcher K.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-MAR-2005) School of Biomedical Sciences, University of
            Ulster, Cromore Road, Coleraine BT52 1SA, UK
REFERENCE   3  (bases 1 to 8008)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8008)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-MAR-2005) School of Biomedical Sciences, University of Ulster, 
            Cromore Road, Coleraine BT52 1SA, UK"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             209..1300
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEALRYPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPGFAKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     1971..1987
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2033..2253
                     /label=URA3 promoter
     CDS             2254..3054
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        3255..3359
                     /label=AmpR promoter
     CDS             3360..4217
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDVGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4391..4979
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        5008..5409
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             5421..5438
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             5448..6092
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDH"
     CDS             6096..6116
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     terminator      6299..6486
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     rep_origin      6497..7843
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 tatactatct gtttcaggga aaactcccag gttcggatgt tcaaaattca atgatgggta
       61 acaagtacga tcgtaaatct gtaaaacagt ttgtcggata ttaggctgta tctcctcaaa
      121 gcgtattcga atatcattga gaagctgcat tttttttttt tttttttttt tttttttttt
      181 atatatattt caaggatata ccattgtaat gtctgcccct aagaagatcg tcgttttgcc
      241 aggtgaccac gttggtcaag aaatcacagc cgaagccatt aaggttctta aagctatttc
      301 tgatgttcgt tccaatgtca agttcgattt cgaaaatcat ttaattggtg gtgctgctat
      361 cgatgctaca ggtgtcccac ttccagatga ggcgctggaa gcctccaaga aggttgatgc
      421 cgttttgtta ggtgctgtgg gtggtcctaa atggggtacc ggtagtgtta gacctgaaca
      481 aggtttacta aaaatccgta aagaacttca attgtacgcc aacttaagac catgtaactt
      541 tgcatccgac tctcttttag acttatctcc aatcaagcca caatttgcta aaggtactga
      601 cttcgttgtt gtcagagaat tagtgggagg tatttacttt ggtaagagaa aggaagacga
      661 tggtgatggt gtcgcttggg atagtgaaca atacaccgtt ccagaagtgc aaagaatcac
      721 aagaatggcc gctttcatgg ccctacaaca tgagccacca ttgcctattt ggtccttgga
      781 taaagctaat gttttggcct cttcaagatt atggagaaaa actgtggagg aaaccatcaa
      841 gaacgaattc cctacattga aggttcaaca tcaattgatt gattctgccg ccatgatcct
      901 agttaagaac ccaacccacc taaatggtat tataatcacc agcaacatgt ttggtgatat
      961 catctccgat gaagccttac gttatccagg ttccttgggt ttgttgccat ctgcgtcctt
     1021 ggcctctttg ccagacaaga acaccgcatt tggtttgtac gaaccatgcc acggttctgc
     1081 tcccggattt gccaagaata aggttgaccc tatcgccact atcttgtctg ctgcaatgat
     1141 gttgaaattg tcattgaact tgcctgaaga aggtaaggcc attgaagatg cagttaaaaa
     1201 ggttttggat gcaggtatca gaactggtga tttaggtggt tccaacagta ccaccgaagt
     1261 cggtgatgct gtcgccgaag aagttaagaa aatccttgct taaaaagatt ctctttttta
     1321 tgatatttgt acataaactt tataaatgaa attcataata gaaacgacac gaaattacaa
     1381 aatggaatat gttcataggg tagacaaacg caatctacat acatttatca agaaggagaa
     1441 aaaggaggat agctaaagga atacaggtaa gcaaattgat actaatggct caacgtgata
     1501 aggaaagaga attgcacttt aacattaata ttgacaagga ggagggcacc acacaaaaag
     1561 ttaggtataa cagaaaatca tgaaactacg aatccctaat ttgatattgg aggattttct
     1621 ctaaaaaaaa aaaaatacaa caaataaaaa acactcaaat gacctgacca tttgatggag
     1681 tttaagtcaa taccttcttg aaccattttc cataatggtg aaagttccct caagaatttt
     1741 actctgtcag aaacggcctt aacgacgtac cggatctatg cggtgtgaaa taccgcacag
     1801 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctac gcaactgttg
     1861 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaagg ggggatgtgc
     1921 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     1981 ggccagtgcc aagcttgcat gcctgcaggt cgacctgcag gcatgcaagc ttttcaattc
     2041 aattcatcat ttttttttta ttcttttttt tgatttcggt ttctttgaaa tttttttgat
     2101 tcggtaatct ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat
     2161 atacgcatat gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac
     2221 agaacaaaaa cctgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt
     2281 gctgctactc atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa
     2341 acaaacttgt gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa
     2401 gcattaggtc ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttca
     2461 atggagggca cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc
     2521 gaagacagaa aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta
     2581 tacagaatag cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt
     2641 gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg
     2701 ttagcagaat tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt
     2761 gacattgcga agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt
     2821 ggaagagatg aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag
     2881 ggagacgcat tgggtcaaca gtatagaacc gtggatgatg tggtctctac aggatctgac
     2941 attattattg ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa
     3001 cgttacagaa aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa
     3061 actgtattat aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta
     3121 tatcagttat tacccgggta ccgagctcga attcttgaag acgaaagggc ctcgtgatac
     3181 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     3241 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3301 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3361 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3421 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3481 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3541 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3601 gtattgacgt cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3661 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3721 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3781 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3841 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3901 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3961 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4021 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4081 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4141 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4201 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4261 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4321 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4381 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4441 caccgctacc agcggtggtt tgtttgccgg atcaagagct atcaactctt tttccgaagg
     4501 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     4561 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4621 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4681 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4741 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4801 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4861 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4921 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4981 acgccagcaa cgcggcccga cctcgacgca tgcaacttct tttctttttt tttcttttct
     5041 ctctcccccg ttgttgtctc accatatccg caatgacaaa aaaaatgatg gaagacacta
     5101 aaggaaaaaa ttaacgacaa agacagcacc aacagatgtc gttgttccag agctgatgag
     5161 gggtatcttc gaacacacga aactttttcc ttccttcatt cacgcacact actctctaat
     5221 gagcaacggt atacggcctt ccttccagtt acttgaattt gaaataaaaa aagtttgccg
     5281 ctttgctatc aagtataaat agacctgcaa ttattaatct tttgtttcct cgtcattgtt
     5341 ctcgttccct ttcttccttg tttctttttc tgcacaatat ttcaagctat accaagcata
     5401 caatcaactc caagcttatg caccatcacc atcaccattc gagcttcatg tcccctatac
     5461 taggttattg gaaaattaag ggccttgtgc aacccactcg acttcttttg gaatatcttg
     5521 aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa
     5581 agtttgaatt gggtttggag tttcccaatc ttccttatta tattgatggt gatgttaaat
     5641 taacacagtc tatggccatc atacgttata tagctgacaa gcacaacatg ttgggtggtt
     5701 gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc ggttttggat attagatacg
     5761 gtgtttcgag aattgcatat agtaaagact ttgaaactct caaagttgat tttcttagca
     5821 agctacctga aatgctgaaa atgttcgaag atcgtttatg tcataaaaca tatttaaatg
     5881 gtgatcatgt aacccatcct gacttcatgt tgtatgacgc tcttgatgtt gttttataca
     5941 tggacccaat gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag
     6001 ctatcccaca aattgataag tacttgaaat ccagcaagta tatagcatgg cctttgcagg
     6061 gctggcaagc cacgtttggt ggtggcgacc atctcgagaa tctttatttt cagggcgcca
     6121 tggtctagca tgactggtgg acagcaaatg ggtcggggat ccagctttgg acttcttcgc
     6181 cagaggtttg gtcaagtctc caatcaaggt tgtcggcttg tctaccttgc cagaaattta
     6241 cgaaaagatg gaaaagggtc aaatcgttgg tagatacgtt gttgacactt ctaaataagc
     6301 gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa taagtgtata
     6361 caaattttaa agtgactctt aggttttaaa acgaaaattc ttgttcttga gtaactcttt
     6421 cctgtaggtc aggttgcttt ctcaggtata gcatgaggtc gctcttattg accacacctc
     6481 taccggcatg cgatatgatc caatatcaaa ggaaatgata gcattgaagg atgagactaa
     6541 tccaattgag gagtggcagc atatagaaca gctaaagggt agtgctgaag gaagcatacg
     6601 ataccccgca tggaatggga taatatcaca ggaggtacta gactaccttt catcctacat
     6661 aaatagacgc atataagtac gcatttaagc ataaacacgc actatgccgt tcttctcatg
     6721 tatatatata tacaggcaac acgcagatat aggtgcgacg tgaacagtga gctgtatgtg
     6781 cgcagctcgc gttgcatttt cggaagcgct cgttttcgga aacgctttga agttcctatt
     6841 ccgaagttcc tattctctag ctagaaagta taggaacttc agagcgcttt tgaaaaccaa
     6901 aagcgctctg aagacgcact ttcaaaaaac caaaaacgca ccggactgta acgagctact
     6961 aaaatattgc gaataccgct tccacaaaca ttgctcaaaa gtatctcttt gctatatatc
     7021 tctgtgctat atccctatat aacctaccca tccacctttc gctccttgaa cttgcatcta
     7081 aactcgacct ctacattttt tatgtttatc tctagtatta ctctttagac aaaaaaattg
     7141 tagtaagaac tattcataga gtgaatcgaa aacaatacga aaatgtaaac atttcctata
     7201 cgtagtatat agagacaaaa tagaagaaac cgttcataat tttctgacca atgaagaatc
     7261 atcaacgcta tcactttctg ttcacaaagt atgcgcaatc cacatcggta tagaatataa
     7321 tcggggatgc ctttatcttg aaaaaatgca cccgcagctt cgctagtaat cagtaaacgc
     7381 gggaagtgga gtcaggcttt ttttatggaa gagaaaatag acaccaaagt agccttcttc
     7441 taaccttaac ggacctacag tgcaaaaagt tatcaagaga ctgcattata gagcgcacaa
     7501 aggagaaaaa aagtaatcta agatgctttg ttagaaaaat agcgctctcg ggatgcattt
     7561 ttgtagaaca aaaaagaagt atagattctt tgttggtaaa atagcgctct cgcgttgcat
     7621 ttctgttctg taaaaatgca gctcagattc tttgtttgaa aaattagcgc tctcgcgttg
     7681 catttttgtt ttacaaaaat gaagcacaga ttcttcgttg gtaaaatagc gctttcgcgt
     7741 tgcatttctg ttctgtaaaa atgcagctca gattctttgt ttgaaaaatt agcgctctcg
     7801 cgttgcattt ttgttctaca aaatgaagca cagatgcttc gttaacaaag atatgctatt
     7861 gaagtgcaag atggaaacgc agaaaatgaa ccggggatgc gacgtgcaag attacctatg
     7921 caatagatgc aatagtttct ccaggaaccg aaatacatac attgtcttcc gtaaagcgct
     7981 agactatata ttattataca ggttcaaa
//
