LOCUS       sequence-47103-e        4443 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 4443)
  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 4443)
  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 3090-06
FEATURES             Location/Qualifiers
     source          1..4443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             87..797
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(232..250)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     500..519
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        869..1089
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             1090..1893
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     primer_bind     complement(2080..2099)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2081..2099)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2186..2203)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2357..2945)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2437..2456)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3116..3907,3908..3976))
                     /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     3739..3758
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3977..4081)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4149..4167
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4205..4227)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        join(4427..4443,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4427..4443,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggcttaaga
       61 tcggtgacgg tgctggttta attaacatgg tgagcaaggg cgaggaggat aacatggcca
      121 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
      181 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
      241 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
      301 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
      361 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
      421 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
      481 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
      541 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
      601 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
      661 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
      721 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
      781 acgagctgta caagtagggc gcgccagatc tggggctggc ttaactatgc ggcatcagag
      841 cagattgtac tgagagtgca ccacgctttt caattcaatt catcattttt tttttattct
      901 tttttttgat ttcggtttct ttgaaatttt tttgattcgg taatctccga acagaaggaa
      961 gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa
     1021 acatgaaatt gcccagtatt cttaacccaa ctgcacagaa caaaaacctg caggaaacga
     1081 agataaatca tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt
     1141 gctgccaagc tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat
     1201 gttcgtacca ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta
     1261 ctaaaaacac atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta
     1321 aaggcattat ccgccaagta caatttttta ctcttcgaag acagaaaatt tgctgacatt
     1381 ggtaatacag tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac
     1441 attacgaatg cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggca
     1501 gaagaagtaa caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc
     1561 tccctatcta ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat
     1621 tttgttatcg gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg
     1681 ttgattatga cacccggtgt gggtttagat gacaagggag acgcattggg tcaacagtat
     1741 agaaccgtgg atgatgtggt ctctacagga tctgacatta ttattgttgg aagaggacta
     1801 tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa
     1861 gcatatttga gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt
     1921 atactaaact cacaaattag agcttcaatt taattatatc agttattacc ctgcggtgtg
     1981 aaataccgca cagatgcgta aggagaaaat acccgagctc gaattcatcg atgatatcag
     2041 atccactagt ggcctatgcg gccgcggatc tgccggtctc cctatagtga gtcgtattaa
     2101 tttcgataag ccaggttaac ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2161 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2221 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2281 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2341 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2401 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2461 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2521 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     2581 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2641 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2701 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2761 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     2821 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2881 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2941 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3001 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3061 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3121 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3181 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3241 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3301 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3361 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3421 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3481 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3541 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3601 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3661 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3721 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3781 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3841 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     3901 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     3961 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4021 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4081 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     4141 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     4201 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     4261 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     4321 actatgcggc atcagagcag attgtactga gagtgcacca tatggacata ttgtcgttag
     4381 aacgcggcta caattaatac ataaccttat gtatcataca catacgattt aggtgacact
     4441 ata
//
