LOCUS       40924_19276        4331 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Plasmid with a TRP1 marker for adding a C-terminal GFP tag or 
            truncating a gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4331)
  AUTHORS   Longtine MS, McKenzie A, Demarini DJ, Shah NG, Wach A, Brachat A, 
            Philippsen P, Pringle JR.
  TITLE     Additional modules for versatile and economical PCR-based gene 
            deletion and modification in Saccharomyces cerevisiae.
  JOURNAL   Yeast 1998;14:953-61.
  PUBMED    9717241
REFERENCE   2  (bases 1 to 4331)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4331)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Yeast"; 
            date: "1998"; volume: "14"; pages: "953-61"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             63..773
                     /codon_start=1
                     /label=GFP (S65T)
                     /note="S65T variant of Aequorea victoria green fluorescent 
                     protein (Heim et al., 1995)"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     terminator      797..984
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     promoter        992..1139
                     /label=TRP1 promoter
     CDS             1140..1811
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(1969..1987)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2245..2833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3007..3864)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3865..3969)
                     /label=AmpR promoter
     promoter        4315..4331
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acagtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg
      121 atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa
      181 aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg ccaacacttg
      241 tcactacttt tacgtatggt gttcaatgct tttcaagata cccagatcat atgaaacggc
      301 atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca
      361 aagatgacgg gaactacaag acacgtgctg aagtcaagtt tgaaggtgat acccttgtta
      421 atagaatcga gttaaaaggt attgatttta aagaagatgg aaacattctt ggacacaaat
      481 tggaatacaa ctataactca cacaatgtat acatcatggc agacaaacaa aagaatggaa
      541 tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa ctagcagacc
      601 attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac aaccattacc
      661 tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa gagagaccac atggtccttc
      721 ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac aaatagggcg
      781 cgccacttct aaataagcga atttcttatg atttatgatt tttattatta aataagttat
      841 aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt
      901 attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagc atgaggtcgc
      961 tcttattgac cacacctcta ccggcagatc tgtacaatct tgatccggag cttttctttt
     1021 tttgccgatt aagaattaat tcggtcgaaa aaagaaaagg agagggccaa gagggagggc
     1081 attggtgact attgagcacg tgagtatacg tgattaagca cacaaaggca gcttggagta
     1141 tgtctgttat taatttcaca ggtagttctg gtccattggt gaaagtttgc ggcttgcaga
     1201 gcacagaggc cgcagaatgt gctctagatt ccgatgctga cttgctgggt attatatgtg
     1261 tgcccaatag aaagagaaca attgacccgg ttattgcaag gaaaatttca agtcttgtaa
     1321 aagcatataa aaatagttca ggcactccga aatacttggt tggcgtgttt cgtaatcaac
     1381 ctaaggagga tgttttggct ctggtcaatg attacggcat tgatatcgtc caactgcatg
     1441 gagatgagtc gtggcaagaa taccaagagt tcctcggttt gccagttatt aaaagactcg
     1501 tatttccaaa agactgcaac atactactca gtgcagcttc acagaaacct cattcgttta
     1561 ttcccttgtt tgattcagaa gcaggtggga caggtgaact tttggattgg aactcgattt
     1621 ctgactgggt tggaaggcaa gagagccccg aaagcttaca ttttatgtta gctggtggac
     1681 tgacgccaga aaatgttggt gatgcgctta gattaaatgg cgttattggt gttgatgtaa
     1741 gcggaggtgt ggagacaaat ggtgtaaaag actctaacaa aatagcaaat ttcgtcaaaa
     1801 atgctaagaa ataggttatt actgagtagt atttatttaa gtattgtttg tgcacttgcc
     1861 tgcggtgtga aataccgcac agatgcgtaa ggaggtttaa acgagctcga attcatcgat
     1921 gatatcagat ccactagtgg cctatgcggc cgcggatctg ccggtctccc tatagtgagt
     1981 cgtattaatt tcgataagcc aggttaacct gcattaatga atcggccaac gcgcggggag
     2041 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     2101 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2161 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2221 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2281 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2341 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2401 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     2461 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2521 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2581 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2641 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     2701 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2761 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2821 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2881 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2941 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3001 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3061 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3121 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3181 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3241 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3301 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3361 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3421 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3481 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3541 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3601 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     3661 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     3721 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3781 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     3841 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     3901 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3961 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     4021 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     4081 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     4141 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     4201 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tggacatatt
     4261 gtcgttagaa cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag
     4321 gtgacactat a
//
