LOCUS       sequence-47096-e        4469 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 4469)
  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 4469)
  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 3080-07
FEATURES             Location/Qualifiers
     source          1..4469
                     /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
                     FICTTGKLPVPWPTLVTTLGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSRLSKDPNEKRDHMVLLE
                     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"
     gene            1086..2010
                     /label=bleMX6
                     /note="yeast selectable marker conferring phleomycin 
                     resistance"
     promoter        1086..1429
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1430..1804
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      1813..2010
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2106..2125)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2107..2125)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2212..2229)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2383..2971)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2463..2482)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3142..3933,3934..4002))
                     /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     3765..3784
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4003..4107)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4175..4193
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4231..4253)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4353..4372
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4453..4469,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4453..4469,1)
                     /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 ctactttagg ttatggtttg atgtgttttg
      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 cctatcaatc tagattatcc 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 agatctgttt agcttgcctc gtccccgccg ggtcacccgg
     1081 ccagcgacat ggaggcccag aataccctcc ttgacagtct tgacgtgcgc agctcagggg
     1141 catgatgtga ctgtcgcccg tacatttagc ccatacatcc ccatgtataa tcatttgcat
     1201 ccatacattt tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct
     1261 gcgagcaggg aaacgctccc ctcacagacg cgttgaattg tccccacgcc gcgcccctgt
     1321 agagaaatat aaaaggttag gatttgccac tgaggttctt ctttcatata cttcctttta
     1381 aaatcttgct aggatacagt tctcacatca catccgaaca taaacaacca tggccaagtt
     1441 gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac
     1501 cgaccggctc gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga
     1561 cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc
     1621 ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac
     1681 gaacttccgg gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg
     1741 ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga
     1801 ctgataatca gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag
     1861 tttttttata ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt
     1921 cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc
     1981 aatcgtatgt gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatccc
     2041 gagctcgaat tcatcgatga tatcagatcc actagtggcc tatgcggccg cggatctgcc
     2101 ggtctcccta tagtgagtcg tattaatttc gataagccag gttaacctgc attaatgaat
     2161 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2221 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2281 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2341 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2401 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2461 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2521 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     2581 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2641 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2701 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2761 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2821 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2881 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2941 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3001 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3061 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3121 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3181 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3241 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3301 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3361 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3421 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3481 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3541 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3601 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3661 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3721 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3781 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3841 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3901 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3961 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4021 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4081 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     4141 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     4201 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     4261 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     4321 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     4381 gcaccatatg gacatattgt cgttagaacg cggctacaat taatacataa ccttatgtat
     4441 catacacata cgatttaggt gacactata
//
