LOCUS       40924_19271        4854 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Plasmid with a kanMX 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 4854)
  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 4854)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4854)
  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..4854
                     /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"
     gene            1031..2387
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     promoter        complement(2492..2510)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2768..3356)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3530..4387)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4388..4492)
                     /label=AmpR promoter
     promoter        4838..4854
                     /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 tgtttagctt gcctcgtccc cgccgggtca
     1021 cccggccagc gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc
     1081 aggggcatga tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt
     1141 tgcatccata cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgca
     1201 gacctgcgag cagggaaacg ctcccctcac agacgcgttg aattgtcccc acgccgcgcc
     1261 cctgtagaga aatataaaag gttaggattt gccactgagg ttcttctttc atatacttcc
     1321 ttttaaaatc ttgctaggat acagttctca catcacatcc gaacataaac aaccatgggt
     1381 aaggaaaaga ctcacgtttc gaggccgcga ttaaattcca acatggatgc tgatttatat
     1441 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat
     1501 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     1561 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     1621 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggcaaa
     1681 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     1741 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     1801 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     1861 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataag
     1921 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     1981 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     2041 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     2101 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     2161 ttgatgctcg atgagttttt ctaatcagta ctgacaataa aaagattctt gttttcaaga
     2221 acttgtcatt tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg
     2281 tgatttatat tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga
     2341 aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat
     2401 actaacgccg ccatccagtt taaacgagct cgaattcatc gatgatatca gatccactag
     2461 tggcctatgc ggccgcggat ctgccggtct ccctatagtg agtcgtatta atttcgataa
     2521 gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2581 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2641 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2701 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2761 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2821 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2881 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2941 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg
     3001 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3061 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3121 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3181 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     3241 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3301 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3361 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3421 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3481 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3541 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3601 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3661 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3721 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3781 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3841 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3901 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     3961 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4021 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4081 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4141 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4201 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4261 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4321 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4381 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4441 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4501 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     4561 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     4621 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     4681 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     4741 catcagagca gattgtactg agagtgcacc atatggacat attgtcgtta gaacgcggct
     4801 acaattaata cataacctta tgtatcatac acatacgatt taggtgacac tata
//
