LOCUS       sequence-6731-ex        4894 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 4894)
  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 4894)
  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: pKT0103
FEATURES             Location/Qualifiers
     source          1..4894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            45..1401
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        45..388
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             389..1198
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(456..475)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1204..1401
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1505..1524)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1506..1524)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1611..1628)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1782..2370)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1862..1881)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2541..3332,3333..3401))
                     /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     3164..3183
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3402..3506)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3574..3592
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3630..3652)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3752..3771
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3852..3870
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3852..3869
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             3940..4656
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      4677..4864
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4732..4756)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
ORIGIN
        1 gatctgttta gcttgcctcg tccccgccgg gtcacccggc cagcgacatg gaggcccaga
       61 ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt
      121 acatttagcc catacatccc catgtataat catttgcatc catacatttt gatggccgca
      181 cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga aacgctcccc
      241 tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata aaaggttagg
      301 atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta ggatacagtt
      361 ctcacatcac atccgaacat aaacaaccat gggtaaggaa aagactcacg tttcgaggcc
      421 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
      481 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
      541 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
      601 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
      661 tgcatggtta ctcaccactg cgatccccgg caaaacagca ttccaggtat tagaagaata
      721 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
      781 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
      841 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
      901 gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt
      961 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     1021 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     1081 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     1141 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     1201 agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata gtttttttat
     1261 attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt tcgcctcgac
     1321 atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg
     1381 tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc agtttaaacg
     1441 agctcgaatt catcgatgat atcagatcca ctagtggcct atgcggccgc ggatctgccg
     1501 gtctccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca ttaatgaatc
     1561 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     1621 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1681 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1741 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1801 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1861 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1921 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     1981 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2041 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2101 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2161 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2221 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2281 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2341 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2401 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2461 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2521 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2581 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2641 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     2701 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2761 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2821 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     2881 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     2941 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3001 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3061 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3121 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3181 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3241 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3301 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3361 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3421 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3481 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     3541 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     3601 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     3661 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     3721 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     3781 caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac cttatgtatc
     3841 atacacatac gatttaggtg acactataga acgcggccgc cagctgaagc ttcgtacgct
     3901 gcaggtcgac ggatcggtga cggtgctggt ttaattaaca tgtctaaagg tgaagaatta
     3961 ttcactggtg ttgtcccaat tttggttgaa ttagatggtg atgttaatgg tcacaaattt
     4021 tctgtctccg gtgaaggtga aggtgatgct acttacggta aattgacctt aaaattgatt
     4081 tgtactactg gtaaattgcc agttccatgg ccaaccttag tcactacttt aggttatggt
     4141 ttgcaatgtt ttgctagata cccagatcat atgaaacaac atgacttttt caagtctgcc
     4201 atgccagaag gttatgttca agaaagaact atttttttca aagatgacgg taactacaag
     4261 accagagctg aagtcaagtt tgaaggtgat accttagtta atagaatcga attaaaaggt
     4321 attgatttta aagaagatgg taacatttta ggtcacaaat tggaatacaa ctataactct
     4381 cacaatgttt acatcactgc tgacaaacaa aagaatggta tcaaagctaa cttcaaaatt
     4441 agacacaaca ttgaagatgg tggtgttcaa ttagctgacc attatcaaca aaatactcca
     4501 attggtgatg gtccagtctt gttaccagac aaccattact tatcctatca atctgcctta
     4561 tccaaagatc caaacgaaaa gagagaccac atggtcttgt tagaatttgt tactgctgct
     4621 ggtattaccc atggtatgga tgaattgtac aaataaggcg cgccacttct aaataagcga
     4681 atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca
     4741 aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc
     4801 tgtaggtcag gttgctttct caggtatagt atgaggtcgc tcttattgac cacacctcta
     4861 ccggcagatc cgctagggat aacagggtaa tata
//
