LOCUS       sequence-47089-e        5698 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 5698)
  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 5698)
  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 3040-05
FEATURES             Location/Qualifiers
     source          1..5698
                     /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        1288..1695
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             1696..2790
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(3335..3354)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3336..3354)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3441..3458)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3612..4200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3692..3711)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4371..5162,5163..5231))
                     /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     4994..5013
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5232..5336)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5404..5422
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5460..5482)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5582..5601
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(5682..5698,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(5682..5698,1)
                     /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 agatctctcg aggagaactt ctagtatatc tacataccta
     1081 atattattgc cttattaaaa atggaatccc aacaattaca tcaaaatcca cattctcttc
     1141 aaaatcaatt gtcctgtact tccttgttca tgtgtgttca aaaacgttat atttatagga
     1201 taattatact ctatttctca acaagtaatt ggttgtttgg ccgagcggtc taaggcgcct
     1261 gattcaagaa atatcttgac cgcagttaac tgtgggaata ctcaggtatc gtaagatgca
     1321 agagttcgaa tctcttagca accattattt ttttcctcaa cataacgaga acacacaggg
     1381 gcgctatcgc acagaatcaa attcgatgac tggaaatttt ttgttaattt cagaggtcgc
     1441 ctgacgcata tacctttttc aactgaaaaa ttgggagaaa aaggaaaggt gagagcgccg
     1501 gaaccggctt ttcatataga atagagaagc gttcatgact aaatgcttgc atcacaatac
     1561 ttgaagttga caatattatt taaggaccta ttgttttttc caataggtgg ttagcaatcg
     1621 tcttactttc taacttttct taccttttac atttcagcaa tatatatata tatatttcaa
     1681 ggatatacca ttctaatgtc tgcccctaag aagatcgtcg ttttgccagg tgaccacgtt
     1741 ggtcaagaaa tcacagccga agccattaag gttcttaaag ctatttctga tgttcgttcc
     1801 aatgtcaagt tcgatttcga aaatcattta attggtggtg ctgctatcga tgctacaggt
     1861 gttccacttc cagatgaggc gctggaagcc tccaagaagg ctgatgccgt tttgttaggt
     1921 gctgtgggtg gtcctaaatg gggtaccggt agtgttagac ctgaacaagg tttactaaaa
     1981 atccgtaaag aacttcaatt gtacgccaac ttaagaccat gtaactttgc atccgactct
     2041 cttttagact tatctccaat caagccacaa tttgctaaag gtactgactt cgttgttgtc
     2101 agagaattag tgggaggtat ttactttggt aagagaaagg aagacgatgg tgatggtgtc
     2161 gcttgggata gtgaacaata caccgttcca gaagtgcaaa gaatcacaag aatggccgct
     2221 ttcatggccc tacaacatga gccaccattg cctatttggt ccttggataa agctaatgtt
     2281 ttggcctctt caagattatg gagaaaaact gtggaggaaa ccatcaagaa cgaattccct
     2341 acattgaagg ttcaacatca attgattgat tctgccgcca tgatcctagt taagaaccca
     2401 acccacctaa atggtattat aatcaccagc aacatgtttg gtgatatcat ctccgatgaa
     2461 gcctccgtta tcccaggttc cttgggtttg ttgccatctg cgtccttggc ctctttgcca
     2521 gacaagaaca ccgcatttgg tttgtacgaa ccatgccacg gttctgctcc agatttgcca
     2581 aagaataagg tcaaccctat cgccactatc ttgtctgctg caatgatgtt gaaattgtca
     2641 ttgaacttgc ctgaagaagg taaggccatt gaagatgcag ttaaaaaggt tttggatgca
     2701 ggtatcagaa ctggtgattt aggtggttcc aacagtacca ccgaagtcgg tgatgctgtc
     2761 gccgaagaag ttaagaaaat ccttgcttaa aaagattctc tttttttatg atatttgtac
     2821 ataaacttta taaatgaaat tcataataga aacgacacga aattacaaaa tggaatatgt
     2881 tcatagggta gacgaaacta tatacgcaat ctacatacat ttatcaagaa ggagaaaaag
     2941 gaggatgtaa aggaatacag gtaagcaaat tgatactaat ggctcaacgt gataaggaaa
     3001 aagaattgca ctttaacatt aatattgaca aggaggaggg caccacacaa aaagttaggt
     3061 gtaacagaaa atcatgaaac tatgattcct aatttatata ttggaggatt ttctctaaaa
     3121 aaaaaaaaat acaacaaata aaaaacactc aatgacctga ccatttgatg gagtttaagt
     3181 caataccttc ttgaaccatt tcccataatg gtgaaagttc cctcaagaat tttactctgt
     3241 cagaaacggc cttaacgacg tagtcgatcg agctcgaatt catcgatgat atcagatcca
     3301 ctagtggcct atgcggccgc ggatctgccg gtctccctat agtgagtcgt attaatttcg
     3361 ataagccagg ttaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3421 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3481 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3541 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3601 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3661 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3721 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3781 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     3841 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3901 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3961 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4021 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     4081 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4141 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4201 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4261 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4321 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4381 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4441 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4501 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4561 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4621 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4681 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4741 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4801 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4861 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4921 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4981 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5041 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5101 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5161 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5221 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5281 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5341 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     5401 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     5461 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     5521 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     5581 gcggcatcag agcagattgt actgagagtg caccatatgg acatattgtc gttagaacgc
     5641 ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg acactata
//
