LOCUS       sequence-47088-e        4574 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 4574)
  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 4574)
  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 3040-04
FEATURES             Location/Qualifiers
     source          1..4574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             87..803
                     /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
                     FVTAAGITHGIDELYK"
     terminator      824..1011
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(879..903)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1101..1381
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             1382..2056
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     primer_bind     complement(2211..2230)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2212..2230)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2317..2334)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2488..3076)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2568..2587)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3247..4038,4039..4107))
                     /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     3870..3889
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4108..4212)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4280..4298
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4336..4358)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        join(4558..4574,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4558..4574,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 acggtgacgg tgctggttta attaacatgt ctaaaggtga agaattattc actggtgttg
      121 tcccaatttt ggttgaatta gatggtgatg ttaatggtca caaattttct gtctccggtg
      181 aaggtgaagg tgatgctact tacggtaaat tgaccttaaa atttatttgt actactggta
      241 aattgccagt tccatggcca accttagtca ctactttaac ttatggtgtt caatgttttt
      301 ctagataccc agatcatatg aaacaacatg actttttcaa gtctgccatg ccagaaggtt
      361 atgttcaaga aagaactatt tttttcaaag atgacggtaa ctacaagacc agagctgaag
      421 tcaagtttga aggtgatacc ttagttaata gaatcgaatt aaaaggtatt gattttaaag
      481 aagatggtaa cattttaggt cacaaattgg aatacaacta taactctcac aatgtttaca
      541 tcatggctga caaacaaaag aatggtatca aagttaactt caaaattaga cacaacattg
      601 aagatggttc tgttcaatta gctgaccatt atcaacaaaa tactccaatt ggtgatggtc
      661 cagtcttgtt accagacaac cattacttat ccactcaatc tgccttatcc aaagatccaa
      721 acgaaaagag agaccacatg gtcttgttag aatttgttac tgctgctggt attacccatg
      781 gtattgatga attgtacaaa taaggcgcgc cacttctaaa taagcgaatt tcttatgatt
      841 tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga
      901 ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt
      961 gctttctcag gtatagtatg aggtcgctct tattgaccac acctctaccg gcagatccgc
     1021 tagggataac agggtaatat agatctgggg ctggcttaac tatgcggcat cagagcagat
     1081 tgtactgaga gtgcaccata aacgacatta ctatatatat aatataggaa gcatttaata
     1141 gacagcatcg taatatatgt gtactttgca gttatgacgc cagatggcag tagtggaaga
     1201 tattctttat tgaaaaatag cttgtcacct tacgtacaat cttgatccgg agcttttctt
     1261 tttttgccga ttaagaatta attcggtcga aaaaagaaaa ggagagggcc aagagggagg
     1321 gcattggtga ctattgagca cgtgagtata cgtgattaag cacacaaagg cagcttggag
     1381 tatgtctgtt attaatttca caggtagttc tggtccattg gtgaaagttt gcggcttgca
     1441 gagcacagag gccgcagaat gtgctctaga ttccgatgct gacttgctgg gtattatatg
     1501 tgtgcccaat agaaagagaa caattgaccc ggttattgca aggaaaattt caagtcttgt
     1561 aaaagcatat aaaaatagtt caggcactcc gaaatacttg gttggcgtgt ttcgtaatca
     1621 acctaaggag gatgttttgg ctctggtcaa tgattacggc attgatatcg tccaactgca
     1681 tggagatgag tcgtggcaag aataccaaga gttcctcggt ttgccagtta ttaaaagact
     1741 cgtatttcca aaagactgca acatactact cagtgcagct tcacagaaac ctcattcgtt
     1801 tattcccttg tttgattcag aagcaggtgg gacaggtgaa cttttggatt ggaactcgat
     1861 ttctgactgg gttggaaggc aagagagccc cgaaagctta cattttatgt tagctggtgg
     1921 actgacgcca gaaaatgttg gtgatgcgct tagattaaat ggcgttattg gtgttgatgt
     1981 aagcggaggt gtggagacaa atggtgtaaa agactctaac aaaatagcaa atttcgtcaa
     2041 aaatgctaag aaataggtta ttactgagta gtatttattt aagtattgtt tgtgcacttg
     2101 cctgcggtgt gaaataccgc acagatgcgt aaggagaaaa tacccgagct cgaattcatc
     2161 gatgatatca gatccactag tggcctatgc ggccgcggat ctgccggtct ccctatagtg
     2221 agtcgtatta atttcgataa gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg
     2281 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2341 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2401 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2461 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2521 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2581 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2641 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     2701 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2761 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2821 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2881 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     2941 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3001 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3061 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3121 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3181 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3241 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3301 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3361 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3421 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3481 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3541 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3601 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3661 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3721 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3781 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3841 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     3901 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3961 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4021 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4081 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4141 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4201 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     4261 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     4321 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4381 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4441 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatggacat
     4501 attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
     4561 taggtgacac tata
//
