LOCUS       sequence-47085-e        4902 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 4902)
  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 4902)
  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 3030-02
FEATURES             Location/Qualifiers
     source          1..4902
                     /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
                     FICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYIYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSRLSKDPNEKRDHMVLLE
                     FVTAAGITHGIDELYK"
     gene            857..2432
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        857..1200
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1201..2229
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             1201..2229
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      2235..2432
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2539..2558)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2540..2558)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2645..2662)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2816..3404)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2896..2915)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3575..4366,4367..4435))
                     /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     4198..4217
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4436..4540)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4608..4626
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4664..4686)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4786..4805
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4886..4902,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4886..4902,1)
                     /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 ttggggtgtt 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 aatacattta taactctcac aatgtttaca
      541 tcactgctga caaacaaaag aatggtatca aagctaactt caaaattaga cacaacattg
      601 aagatggttc tgttcaatta gctgaccatt atcaacaaaa tactccaatt ggtgatggtc
      661 cagtcttgtt accagacaac cattacttat ccactcaatc tagattatcc aaagatccaa
      721 acgaaaagag agaccacatg gtcttgttag aatttgttac tgctgctggt attacccatg
      781 gtattgatga attgtacaaa taaggcgcgc cagatctgtt tagcttgcct cgtccccgcc
      841 gggtcacccg gccagcgaca tggaggccca gaataccctc cttgacagtc ttgacgtgcg
      901 cagctcaggg gcatgatgtg actgtcgccc gtacatttag cccatacatc cccatgtata
      961 atcatttgca tccatacatt ttgatggccg cacggcgcga agcaaaaatt acggctcctc
     1021 gctgcagacc tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt gtccccacgc
     1081 cgcgcccctg tagagaaata taaaaggtta ggatttgcca ctgaggttct tctttcatat
     1141 acttcctttt aaaatcttgc taggatacag ttctcacatc acatccgaac ataaacaacc
     1201 atgggtaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc
     1261 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc
     1321 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa
     1381 gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac
     1441 attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg
     1501 ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg
     1561 gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa
     1621 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg
     1681 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat
     1741 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc
     1801 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag
     1861 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg
     1921 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg
     1981 ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt
     2041 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc
     2101 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat
     2161 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca
     2221 aaggaataat cagtactgac aataaaaaga ttcttgtttt caagaacttg tcatttgtat
     2281 agttttttta tattgtagtt gttctatttt aatcaaatgt tagcgtgatt tatatttttt
     2341 ttcgcctcga catcatctgc ccagatgcga agttaagtgc gcagaaagta atatcatgcg
     2401 tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa cgccgccatc
     2461 cagtgtcgaa aacgagctcg aattcatcga tgatatcaga tccactagtg gcctatgcgg
     2521 ccgcggatct gccggtctcc ctatagtgag tcgtattaat ttcgataagc caggttaacc
     2581 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2641 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2701 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2761 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2821 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2881 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2941 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3001 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3061 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3121 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3181 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3241 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3301 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3361 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3421 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3481 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3541 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3601 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3661 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3721 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3781 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3841 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3901 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3961 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4021 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4081 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4141 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4201 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4261 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4321 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4381 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4441 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4501 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4561 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     4621 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     4681 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     4741 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     4801 ttgtactgag agtgcaccat atggacatat tgtcgttaga acgcggctac aattaataca
     4861 taaccttatg tatcatacac atacgattta ggtgacacta ta
//
