LOCUS       sequence-6749-ex        4901 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 4901)
  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 4901)
  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: pKT0239
FEATURES             Location/Qualifiers
     source          1..4901
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        45..388
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             389..1087
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSNRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1091..1288
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1392..1411)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1393..1411)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1498..1515)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1669..2257)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1749..1768)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2428..3219,3220..3288))
                     /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     3051..3070
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3289..3393)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3461..3479
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3517..3539)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3639..3658
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3739..3757
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3739..3756
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             3827..4540
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             4562..4651
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     terminator      4684..4871
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4739..4763)
                     /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 ggcagaacca gcccaaaaaa agcaaaaaca
      421 aactgttcag gagcgcaagg cgtttatctc ccgtatcact aatgaaacta aaattcaaat
      481 cgctatttcg ctgaatggtg gttatattca aataaaagat tcgattcttc ctgcaaagaa
      541 ggatgacgat gtagcttccc aagctactca gtcacaggtc atcgatattc acacaggtgt
      601 tggctttttg gatcatatga tccatgcgtt ggcaaaacac tctggttggt ctcttattgt
      661 tgaatgtatc ggtgacctgc acattgacga tcaccatact accgaagatt gcggtatcgc
      721 attagggcaa gcgttcaaag aagcaatggg tgctgtccgt ggtgtaaaaa gattcggtac
      781 tgggttcgca ccattggatg aggcgctatc acgtgccgta gtcgatttat ctaatagacc
      841 atttgctgta atcgaccttg gattgaagag agagatgatt ggtgatttat ccactgaaat
      901 gattccacac tttttggaaa gtttcgcgga ggcggccaga attactttgc atgttgattg
      961 tctgagaggt ttcaacgatc accacagaag tgagagtgcg ttcaaggctt tggctgttgc
     1021 cataagagaa gctatttcta gcaatggcac caatgacgtt ccctcaacca aaggtgtttt
     1081 gatgtgaagt actgacaata aaaagattct tgttttcaag aacttgtcat ttgtatagtt
     1141 tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata ttttttttcg
     1201 cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa
     1261 tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc gccatccagt
     1321 ttaaacgagc tcgaattcat cgatgatatc agatccacta gtggcctatg cggccgcgga
     1381 tctgccggtc tccctatagt gagtcgtatt aatttcgata agccaggtta acctgcatta
     1441 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     1501 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1561 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1621 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1681 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1741 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1801 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1861 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1921 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1981 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2041 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2101 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2161 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2221 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2281 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2341 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2401 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2461 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     2521 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     2581 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     2641 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     2701 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     2761 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     2821 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     2881 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     2941 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3001 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3061 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     3121 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     3181 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     3241 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3301 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3361 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     3421 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     3481 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     3541 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     3601 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     3661 gagagtgcac catatggaca tattgtcgtt agaacgcggc tacaattaat acataacctt
     3721 atgtatcata cacatacgat ttaggtgaca ctatagaacg cggccgccag ctgaagcttc
     3781 gtacgctgca ggtcgacgga tcggtgacgg tgctggttta attaacatgt ctaaaggtga
     3841 agaattattc actggtgttg tcccaatttt ggttgaatta gatggtgatg ttaatggtca
     3901 caaattttct gtctccggtg aaggtgaagg tgatgctact tacggtaaat tgaccttaaa
     3961 atttatttgt actactggta aattgccagt tccatggcca accttagtca ctactttagg
     4021 ttatggtttg atgtgttttg ctagataccc agatcatatg aaacaacatg actttttcaa
     4081 gtctgccatg ccagaaggtt atgttcaaga aagaactatt tttttcaaag atgacggtaa
     4141 ctacaagacc agagctgaag tcaagtttga aggtgatacc ttagttaata gaatcgaatt
     4201 aaaaggtatt gattttaaag aagatggtaa cattttaggt cacaaattgg aatacaacta
     4261 taactctcac aatgtttaca tcatggctga caaacaaaag aatggtatca aagttaactt
     4321 caaaattaga cacaacattg aagatggttc tgttcaatta gctgaccatt atcaacaaaa
     4381 tactccaatt ggtgatggtc cagtcttgtt accagacaac cattacttat cctatcaatc
     4441 tgccttatcc aaagatccaa acgaaaagag agaccacatg gtcttgttag aatttgttac
     4501 tgctgctggt attacccatg gtatggatga attgtacaaa cccgggttaa ttaacatctt
     4561 ttacccatac gatgttcctg actatgcggg ctatccgtat gacgtcccgg actatgcagg
     4621 atcctatcca tatgacgttc cagattacgc tgctcagtgc tgaggcgcgc cacttctaaa
     4681 taagcgaatt tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt
     4741 gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac
     4801 tctttcctgt aggtcaggtt gctttctcag gtatagtatg aggtcgctct tattgaccac
     4861 acctctaccg gcagatccgc tagggataac agggtaatat a
//
