LOCUS       sequence-47091-e        4580 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 4580)
  AUTHORS   Young CL, Raden DL, Caplan JL, Czymmek KJ, Robinson AS
  TITLE     Cassette series designed for live-cell imaging of proteins and 
            high-resolution techniques in yeast.
  JOURNAL   Yeast. 2012 Mar;29(3-4):119-36. doi: 10.1002/yea.2895. Epub 2012 Apr
            4.
  PUBMED    22473760
REFERENCE   2  (bases 1 to 4580)
  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: "2012-03"; volume: "29(3-4):119-36. doi"; pages: " 
            10.1002/yea.2895. Epub 2012 Apr 4"
COMMENT     Sequence Label: pCY 3050-06
FEATURES             Location/Qualifiers
     source          1..4580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(114..142)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     737..757
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      821..1008
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(876..900)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1050..1266
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             1267..2070
                     /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
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLATGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     primer_bind     complement(2217..2236)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2218..2236)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2323..2340)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2494..3082)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2574..2593)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3253..4044,4045..4113))
                     /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     3876..3895
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4114..4218)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4286..4304
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4342..4364)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4464..4483
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4564..4580,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4564..4580,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggcttaaga
       61 tcggtgacgg tgctggttta attaacagta aaggagaaga acttttcact ggagttgtcc
      121 caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg
      181 gtgaaggtga tgcaacatac ggaaaactta cccttaaatt gatttgcact actggaaaac
      241 tacctgttcc atggccaaca cttgtcacta ctttgggtta tggtctaatg tgctttgcta
      301 gatacccaga tcatatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg
      361 tacaggaaag aactatattt ttcaaagatg acgggaacta caagacacgt gctgaagtca
      421 agtttgaagg tgataccctt gttaatagaa tcgagttaaa aggtattgat tttaaagaag
      481 atggaaacat tcttggacac aaattggaat acaactataa ctcacacaat gtatacatca
      541 ctgcagacaa acaaaagaat ggaatcaaag ctaacttcaa aattagacac aacattgaag
      601 atggaggtgt tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg
      661 tccttttacc agacaaccat tacctgtcct atcaatctgc cctttcgaaa gatcccaacg
      721 aaaagagaga ccacatggtc cttcttgagt ttgtaacagc tgctgggatt acacatggca
      781 tggatgaact atacaaatag ggcgcgccac ttctaaataa gcgaatttct tatgatttat
      841 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
      901 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
      961 ttctcaggta tagtatgagg tcgctcttat tgaccacacc tctaccggca gatccgctag
     1021 ggataacagg gtaatataga tctaagcttt tcaattcatc tttttttttt ttgttctttt
     1081 ttttgattcc ggtttctttg aaattttttt gattcggtaa tctccgagca gaaggaagaa
     1141 cgaaggaagg agcacagact tagattggta tatatacgca tatgtggtgt tgaagaaaca
     1201 tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga
     1261 taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct
     1321 gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt
     1381 cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta
     1441 aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag
     1501 gcattatccg ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt
     1561 aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt
     1621 acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcggaa
     1681 gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc
     1741 ctagctactg gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt
     1801 gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg
     1861 attatgacac ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga
     1921 accgtggatg atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt
     1981 gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca
     2041 tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata
     2101 ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccgg ggagctcgaa
     2161 ttcatcgatg atatcagatc cactagtggc ctatgcggcc gcggatctgc cggtctccct
     2221 atagtgagtc gtattaattt cgataagcca ggttaacctg cattaatgaa tcggccaacg
     2281 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     2341 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2401 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2461 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2521 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2581 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2641 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     2701 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2761 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2821 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2881 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     2941 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3001 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3061 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3121 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3181 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3241 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3301 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3361 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3421 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3481 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3541 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3601 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3661 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3721 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3781 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3841 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3901 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3961 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4021 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4081 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4141 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4201 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     4261 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     4321 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     4381 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     4441 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     4501 ggacatattg tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat
     4561 acgatttagg tgacactata
//
