LOCUS       sequence-6739-ex        5444 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 5444)
  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 5444)
  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: pKT0232
FEATURES             Location/Qualifiers
     source          1..5444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             18..515
                     /codon_start=1
                     /product="13 tandem Myc epitope tags"
                     /label=13xMyc
                     /translation="EQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLIS
                     EEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLD
                     GEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISE
                     EDL"
     terminator      573..760
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(628..652)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            835..2191
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        835..1178
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1179..1988
                     /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(1246..1265)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1994..2191
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2295..2314)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2296..2314)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2401..2418)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2572..3160)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2652..2671)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3331..4122,4123..4191))
                     /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     3954..3973
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4192..4296)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4364..4382
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4420..4442)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4542..4561
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4642..4660
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4642..4659
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4730..5443
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYIYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
ORIGIN
        1 ccgggttaat taacggtgaa caaaagctaa tctccgagga agacttgaac ggtgaacaaa
       61 aattaatctc agaagaagac ttgaacggac tcgacggtga acaaaagttg atttctgaag
      121 aagatttgaa cggtgaacaa aagctaatct ccgaggaaga cttgaacggt gaacaaaaat
      181 taatctcaga agaagacttg aacggactcg acggtgaaca aaagttgatt tctgaagaag
      241 atttgaacgg tgaacaaaag ctaatctccg aggaagactt gaacggtgaa caaaaattaa
      301 tctcagaaga agacttgaac ggactcgacg gtgaacaaaa gttgatttct gaagaagatt
      361 tgaacggtga acaaaagcta atctccgagg aagacttgaa cggtgaacaa aaattaatct
      421 cagaagaaga cttgaacgga ctcgacggtg aacaaaagtt gatttctgaa gaagatttga
      481 acggtgaaca aaagctaatc tccgaggaag acttgaacgg tgaacaaaaa ttaatcaatc
      541 actagtgaat tcggcgcgcc acttctaaat aagcgaattt cttatgattt atgattttta
      601 ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac tcttaggttt
      661 taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg ctttctcagg
      721 tatagtatga ggtcgctctt attgaccaca cctctaccgg cagatccgct agggataaca
      781 gggtaatata gatctgttta gcttgcctcg tccccgccgg gtcacccggc cagcgacatg
      841 gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac
      901 tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc catacatttt
      961 gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga
     1021 aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata
     1081 aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta
     1141 ggatacagtt ctcacatcac atccgaacat aaacaaccat gggtaaggaa aagactcacg
     1201 tttcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     1261 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
     1321 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     1381 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     1441 ctcctgatga tgcatggtta ctcaccactg cgatccccgg caaaacagca ttccaggtat
     1501 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     1561 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     1621 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     1681 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac
     1741 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     1801 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     1861 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     1921 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     1981 ttttctaatc agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata
     2041 gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt
     2101 tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt
     2161 caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc
     2221 agtttaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct atgcggccgc
     2281 ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca
     2341 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     2401 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2461 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2521 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2581 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2641 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2701 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2761 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2821 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2881 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2941 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3001 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3061 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3121 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3181 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3241 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3301 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3361 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3421 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3481 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3541 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3601 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3661 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3721 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3781 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3841 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3901 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3961 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4021 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4081 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4141 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4201 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4261 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4321 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     4381 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     4441 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     4501 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     4561 actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac
     4621 cttatgtatc atacacatac gatttaggtg acactataga acgcggccgc cagctgaagc
     4681 ttcgtacgct gcaggtcgac ggatcggtga cggtgctggt ttaattaaca tgtctaaagg
     4741 tgaagaatta ttcactggtg ttgtcccaat tttggttgaa ttagatggtg atgttaatgg
     4801 tcacaaattt tctgtctccg gtgaaggtga aggtgatgct acttacggta aattgacctt
     4861 aaaatttatt tgtactactg gtaaattgcc agttccatgg ccaaccttag tcactacttt
     4921 aacttggggt gttcaatgtt tttctagata cccagatcat atgaaacaac atgacttttt
     4981 caagtctgcc atgccagaag gttatgttca agaaagaact atttttttca aagatgacgg
     5041 taactacaag accagagctg aagtcaagtt tgaaggtgat accttagtta atagaatcga
     5101 attaaaaggt attgatttta aagaagatgg taacatttta ggtcacaaat tggaatacat
     5161 ttataactct cacaatgttt acatcactgc tgacaaacaa aagaatggta tcaaagctaa
     5221 cttcaaaatt agacacaaca ttgaagatgg ttctgttcaa ttagctgacc attatcaaca
     5281 aaatactcca attggtgatg gtccagtctt gttaccagac aaccattact tatccactca
     5341 atctgcctta tccaaagatc caaacgaaaa gagagaccac atggtcttgt tagaatttgt
     5401 tactgctgct ggtattaccc atggtatgga tgaattgtac aaac
//
