LOCUS       sequence-6733-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: pKT0174
FEATURES             Location/Qualifiers
     source          1..4900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      19..206
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(74..98)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        281..624
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      1443..1640
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1747..1766)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1748..1766)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1853..1870)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2024..2612)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2104..2123)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2783..3574,3575..3643))
                     /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     3406..3425
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3644..3748)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3816..3834
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3872..3894)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3994..4013
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4094..4112
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4094..4111
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4182..4898
                     /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 cgcgccactt ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt
       61 ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc
      121 ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc
      181 gctcttattg accacacctc taccggcaga tccgctaggg ataacagggt aatatagatc
      241 tgtttagctt gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac
      301 cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat
      361 ttagcccata catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc
      421 gcgaagcaaa aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac
      481 agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt
      541 gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca
      601 catcacatcc gaacataaac aaccatgaca gtcaacacta agacctatag tgagagagca
      661 gaaactcatg cctcaccagt agcacagcga ttatttcgat taatggaact gaagaaaacc
      721 aatttatgtg catcaattga cgttgatacc actaaggaat tccttgaatt aattgataaa
      781 ttaggtcctt atgtatgctt aatcaagact catattgata taatcaatga tttttcctat
      841 gaatccacta ttgaaccatt attagaactt tcacgtaaac atcaatttat gatttttgaa
      901 gatagaaaat ttgctgatat tggtaatacc gtaaagaaac aatatattgg tggagtttat
      961 aaaattagta gttgggcaga tattaccaat gctcatggtg tcactgggaa tggagtggtt
     1021 gaaggattaa aacagggagc taaagaaacc accaccaacc aagagccaag agggttattg
     1081 atgttagctg aattatcatc agtgggatca ttagcatatg gagaatattc tcaaaaaact
     1141 gttgaaattg ctaaatccga taaggaattt gttattggat ttattgccca acgtgatatg
     1201 ggtggccaag aagaaggatt tgattggctt attatgacac ctggagttgg attagatgat
     1261 aaaggtgatg gattaggaca acaatataga actgttgatg aagttgttag cactggaact
     1321 gatattatca ttgttggtag aggattgttt ggtaaaggaa gagatccaga tattgaaggt
     1381 aaaaggtata gaaatgctgg ttggaatgct tatttgaaaa agactggcca attataatca
     1441 gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata
     1501 ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca
     1561 tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt
     1621 gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatcca gtgtcgaaaa
     1681 cgagctcgaa ttcatcgatg atatcagatc cactagtggc ctatgcggcc gcggatctgc
     1741 cggtctccct atagtgagtc gtattaattt cgataagcca ggttaacctg cattaatgaa
     1801 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     1861 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     1921 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     1981 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2041 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2101 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2161 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     2221 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2281 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2341 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2401 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2461 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2521 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2581 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     2641 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2701 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     2761 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     2821 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     2881 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     2941 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3001 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3061 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3121 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3181 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3241 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3301 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     3361 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3421 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3481 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3541 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     3601 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     3661 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     3721 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     3781 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     3841 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     3901 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     3961 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     4021 tgcaccatat ggacatattg tcgttagaac gcggctacaa ttaatacata accttatgta
     4081 tcatacacat acgatttagg tgacactata gaacgcggcc gccagctgaa gcttcgtacg
     4141 ctgcaggtcg acggatcggt gacggtgctg gtttaattaa catgtctaaa ggtgaagaat
     4201 tattcactgg tgttgtccca attttggttg aattagatgg tgatgttaat ggtcacaaat
     4261 tttctgtctc cggtgaaggt gaaggtgatg ctacttacgg taaattgacc ttaaaattta
     4321 tttgtactac tggtaaattg ccagttccat ggccaacctt agtcactact ttaacttggg
     4381 gtgttcaatg tttttctaga tacccagatc atatgaaaca acatgacttt ttcaagtctg
     4441 ccatgccaga aggttatgtt caagaaagaa ctattttttt caaagatgac ggtaactaca
     4501 agaccagagc tgaagtcaag tttgaaggtg ataccttagt taatagaatc gaattaaaag
     4561 gtattgattt taaagaagat ggtaacattt taggtcacaa attggaatac atttataact
     4621 ctcacaatgt ttacatcact gctgacaaac aaaagaatgg tatcaaagct aacttcaaaa
     4681 ttagacacaa cattgaagat ggttctgttc aattagctga ccattatcaa caaaatactc
     4741 caattggtga tggtccagtc ttgttaccag acaaccatta cttatccact caatctgcct
     4801 tatccaaaga tccaaacgaa aagagagacc acatggtctt gttagaattt gttactgctg
     4861 ctggtattac ccatggtatg gatgaattgt acaaataagg
//
