LOCUS       sequence-62061-e        3049 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 3049)
  AUTHORS   Kitada T, Kuryan BG, Tran NN, Song C, Xue Y, Carey M, Grunstein M
  TITLE     Mechanism for epigenetic variegation of gene expression at yeast 
            telomeric heterochromatin.
  JOURNAL   Genes Dev. 2012 Nov 1;26(21):2443-55. doi: 10.1101/gad.201095.112.
  PUBMED    23124068
REFERENCE   2  (bases 1 to 3049)
  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"; doi: 
            "10.1101/gad.201095"; journalName: "Genes Dev"; date: "2012-11-1- 
            1"; volume: "26"; issue: "21"; pages: "2443-55"
COMMENT     Sequence Label: pTK023
FEATURES             Location/Qualifiers
     source          1..3049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             657..1457
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     CDS             1488..2201
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1488..2201
                     /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             2409..2429
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2701..2720)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2702..2720)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2727..2744)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2727..2743)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2736..2758)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2971..2990)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgtttatgag ttcgttcgat ttttttaata ttcataggct ttcttgataa tggcaaccac
       61 ttgttctttg gtgaattgaa ctgggttagt caaatggcag gcatcatgca tggcgtgttc
      121 agccaaaatg tcaaaatctt cggttttaac acctaagtct ttcaagtttc ttggaatgtt
      181 catggttctg tttaaaacgt gcaaagcctt gatggtttct tctggatctt cttgagaagc
      241 accgcaatgc aaggcaattt cacctaatct cttcttggcc tttggacatt gcatgttggc
      301 ctcttgaaca tgaggcaaca agacagcgtt acaaacacca tgaggcaagt ggtagaaacc
      361 accaagttga tgagcaaggg catgaacata acctagagaa gcattgttga aagccatacc
      421 tgccaagtat tctgcgtaac acatatcagt tctggccttc ttgtctttac cgtctttgta
      481 tgcggcgacc aagcttgatt cggtaatctc cgagcagaag gaagaacgaa ggaaggagca
      541 cagacttaga ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt
      601 attcttaacc caactgcaca gaacaaaaac ctgcaggaaa cgaagataaa tccggaatgt
      661 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
      721 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
      781 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
      841 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
      901 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
      961 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
     1021 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
     1081 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
     1141 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
     1201 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     1261 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
     1321 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     1381 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     1441 gatgcggcca gcaaaacggt ggaggtggag gtggaggtgg aatccggatg tctaaaggtg
     1501 aagaattatt cactggtgtt gtcccaattt tggttgaatt agatggtgat gttaatggtc
     1561 acaaattttc tgtctccggt gaaggtgaag gtgatgctac ttacggtaaa ttgaccttaa
     1621 aatttatttg tactactggt aaattgccag ttccatggcc aaccttagtc actactttaa
     1681 cttatggtgt tcaatgtttt tctagatacc cagatcatat gaaacaacat gactttttca
     1741 agtctgccat gccagaaggt tatgttcaag aaagaactat ttttttcaaa gatgacggta
     1801 actacaagac cagagctgaa gtcaagtttg aaggtgatac cttagttaat agaatcgaat
     1861 taaaaggtat tgattttaaa gaagatggta acattttagg tcacaaattg gaatacaact
     1921 ataactctca caatgtttac atcatggctg acaaacaaaa gaatggtatc aaagttaact
     1981 tcaaaattag acacaacatt gaagatggtt ctgttcaatt agctgaccat tatcaacaaa
     2041 atactccaat tggtgatggt ccagtcttgt taccagacaa ccattactta tccactcaat
     2101 ctgccttatc caaagatcca aacgaaaaga gagaccacat ggtcttgtta gaatttgtta
     2161 ctgctgctgg tattacccat ggtatggatg aattgtacaa agcatccaac ttgaacattt
     2221 cgagaaagct taccatatca accccatcat gctctttcga aaattcaaat agcacatcca
     2281 ttccttcgcc cgtttcctca tctcaaagcc acactccaat gagaaacatg agctcactct
     2341 ctgataacag cgttttcagc cggaatatgg aacaatcatc accaatcact ccagcgcgct
     2401 gtactccacc aaagaagaag agaaaggttg cctaagagaa gatgcggcca gcaaaactaa
     2461 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
     2521 ttatatcagt tattaccctc gacggatccc agagtagagg tagatgtgag agagtgtgtg
     2581 ggtgtgggtg tgggtgtgtg ggtgtgtggg tgtgtggatg tgggtgtggt gtgggtgtgg
     2641 tgtgggtgtg gtggcagggg gaattcgcgg ccgcggcgcc aatgcattgg gcccaattcg
     2701 ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa
     2761 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     2821 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2881 tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2941 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3001 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttt
//
