LOCUS       sequence-6742-ex        4900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4900)
  AUTHORS   Sheff MA, Thorn KS
  TITLE     Optimized cassettes for fluorescent protein tagging in Saccharomyces
            cerevisiae.
  JOURNAL   Yeast 2004 Jun;21(8):661-70.
  PUBMED    15197731
REFERENCE   2  (bases 1 to 4900)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Yeast"; 
            date: "2004-06"; volume: "21(8)"; pages: "661-70"
COMMENT     Sequence Label: pKT0209
FEATURES             Location/Qualifiers
     source          1..4900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(61..80)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(62..80)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(167..184)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(338..926)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(418..437)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1097..1888,1889..1957))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1720..1739
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1958..2062)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2130..2148
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2186..2208)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2308..2327
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2408..2426
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2408..2425
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             2496..3212
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             2496..3209
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      3233..3420
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3288..3312)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        3495..3838
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4657..4854
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
ORIGIN
        1 cgaattcatc gatgatatca gatccactag tggcctatgc ggccgcggat ctgccggtct
       61 ccctatagtg agtcgtatta atttcgataa gccaggttaa cctgcattaa tgaatcggcc
      121 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
      181 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
      241 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
      301 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
      361 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
      421 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
      481 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
      541 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
      601 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
      661 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
      721 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
      781 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
      841 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
      901 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
      961 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     1021 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     1081 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     1141 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     1201 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     1261 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     1321 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     1381 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     1441 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     1501 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     1561 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     1621 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     1681 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     1741 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     1801 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     1861 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     1921 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     1981 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     2041 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     2101 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     2161 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     2221 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     2281 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     2341 atatggacat attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac
     2401 acatacgatt taggtgacac tatagaacgc ggccgccagc tgaagcttcg tacgctgcag
     2461 gtcgacggat cggtgacggt gctggtttaa ttaacatgtc taaaggtgaa gaattattca
     2521 ctggtgttgt cccaattttg gttgaattag atggtgatgt taatggtcac aaattttctg
     2581 tctccggtga aggtgaaggt gatgctactt acggtaaatt gaccttaaaa tttatttgta
     2641 ctactggtaa attgccagtt ccatggccaa ccttagtcac tactttaact tatggtgttc
     2701 aatgtttttc tagataccca gatcatatga aacaacatga ctttttcaag tctgccatgc
     2761 cagaaggtta tgttcaagaa agaactattt ttttcaaaga tgacggtaac tacaagacca
     2821 gagctgaagt caagtttgaa ggtgatacct tagttaatag aatcgaatta aaaggtattg
     2881 attttaaaga agatggtaac attttaggtc acaaattgga atacaactat aactctcaca
     2941 atgtttacat catggctgac aaacaaaaga atggtatcaa agttaacttc aaaattagac
     3001 acaacattga agatggttct gttcaattag ctgaccatta tcaacaaaat actccaattg
     3061 gtgatggtcc agtcttgtta ccagacaacc attacttatc cactcaatct gccttatcca
     3121 aagatccaaa cgaaaagaga gaccacatgg tcttgttaga atttgttact gctgctggta
     3181 ttacccatgg tatggatgaa ttgtacaaat aaggcgcgcc acttctaaat aagcgaattt
     3241 cttatgattt atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt
     3301 ttaaagtgac tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta
     3361 ggtcaggttg ctttctcagg tatagtatga ggtcgctctt attgaccaca cctctaccgg
     3421 cagatccgct agggataaca gggtaatata gatctgttta gcttgcctcg tccccgccgg
     3481 gtcacccggc cagcgacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca
     3541 gctcaggggc atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat
     3601 catttgcatc catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc
     3661 tgcagacctg cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg
     3721 cgcccctgta gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac
     3781 ttccttttaa aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaaccat
     3841 gacagtcaac actaagacct atagtgagag agcagaaact catgcctcac cagtagcaca
     3901 gcgattattt cgattaatgg aactgaagaa aaccaattta tgtgcatcaa ttgacgttga
     3961 taccactaag gaattccttg aattaattga taaattaggt ccttatgtat gcttaatcaa
     4021 gactcatatt gatataatca atgatttttc ctatgaatcc actattgaac cattattaga
     4081 actttcacgt aaacatcaat ttatgatttt tgaagataga aaatttgctg atattggtaa
     4141 taccgtaaag aaacaatata ttggtggagt ttataaaatt agtagttggg cagatattac
     4201 caatgctcat ggtgtcactg ggaatggagt ggttgaagga ttaaaacagg gagctaaaga
     4261 aaccaccacc aaccaagagc caagagggtt attgatgtta gctgaattat catcagtggg
     4321 atcattagca tatggagaat attctcaaaa aactgttgaa attgctaaat ccgataagga
     4381 atttgttatt ggatttattg cccaacgtga tatgggtggc caagaagaag gatttgattg
     4441 gcttattatg acacctggag ttggattaga tgataaaggt gatggattag gacaacaata
     4501 tagaactgtt gatgaagttg ttagcactgg aactgatatt atcattgttg gtagaggatt
     4561 gtttggtaaa ggaagagatc cagatattga aggtaaaagg tatagaaatg ctggttggaa
     4621 tgcttatttg aaaaagactg gccaattata atcagtactg acaataaaaa gattcttgtt
     4681 ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat
     4741 gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt
     4801 gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg
     4861 attcgatact aacgccgcca tccagtgtcg aaaacgagct
//
