LOCUS       sequence-47095-e        4909 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 4909)
  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 4909)
  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 3080-01
FEATURES             Location/Qualifiers
     source          1..4909
                     /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..2442
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1086..1429
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1430..2239
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1497..1516)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2245..2442
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2546..2565)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2547..2565)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2652..2669)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2823..3411)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2903..2922)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3582..4373,4374..4442))
                     /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     4205..4224
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4443..4547)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4615..4633
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4671..4693)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4793..4812
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4893..4909,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4893..4909,1)
                     /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 tgggtaagga
     1441 aaagactcac gtttcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     1501 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa
     1561 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     1621 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     1681 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gcaaaacagc
     1741 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     1801 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     1861 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     1921 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataagctttt
     1981 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     2041 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     2101 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     2161 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     2221 gctcgatgag tttttctaat cagtactgac aataaaaaga ttcttgtttt caagaacttg
     2281 tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt tagcgtgatt
     2341 tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc gcagaaagta
     2401 atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa
     2461 cgccgccatc cagtttaaac gagctcgaat tcatcgatga tatcagatcc actagtggcc
     2521 tatgcggccg cggatctgcc ggtctcccta tagtgagtcg tattaatttc gataagccag
     2581 gttaacctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2641 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2701 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2761 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2821 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2881 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2941 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3001 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3061 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3121 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3181 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3241 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     3301 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3361 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3421 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3481 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3541 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3601 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3661 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3721 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3781 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3841 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3901 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3961 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4021 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4081 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4141 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4201 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4261 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4321 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4381 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4441 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4501 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4561 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     4621 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     4681 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     4741 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     4801 gagcagattg tactgagagt gcaccatatg gacatattgt cgttagaacg cggctacaat
     4861 taatacataa ccttatgtat catacacata cgatttaggt gacactata
//
