LOCUS       sequence-8470-ex        5787 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 5787)
  AUTHORS   Di Talia S, Skotheim JM, Bean JM, Siggia ED, Cross FR
  TITLE     The effects of molecular noise and size control on variability in 
            the budding yeast cell cycle.
  JOURNAL   Nature. 2007 Aug 23. 448(7156):947-51.
  PUBMED    17713537
REFERENCE   2  (bases 1 to 5787)
  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: "Nature. 
            2007 Aug 23. 448(7156):947-51."
COMMENT     Sequence Label: pSD02
FEATURES             Location/Qualifiers
     source          1..5787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        196..416
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             417..1220
                     /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"
     rep_origin      complement(1351..1806)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1488..1509)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1700..1719
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1936..1958
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1950..1967
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1951..1967
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1977..1996
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1977..1995
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2020..2036
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2021..2037
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(2105..2292)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     2213..2237
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2313..2990)
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(2375..2398)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2573..2592)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2790..2810
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2842..2860
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(3474..3490)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3474..3490)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3527..3547)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3527..3545)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3566..3582)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3566..3582)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3579..3601)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3590..3606
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3614..3620,3621..3638,3639..3644))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3797..3814)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3968..4556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4048..4067)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4727..5518,5519..5587))
                     /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     5350..5369
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5588..5692)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5760..5778
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
      361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
      421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
      481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
      541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
      601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
      661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
      721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
      781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
      841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
      901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
      961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
     1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
     1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat
     1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1801 gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     1861 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     1921 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
     1981 acgactcact atagggcgaa ttgggtaccg ggccccccct cgaggtcgac ggtatcgata
     2041 agcttgatat cgaattcctg cagcccgggg gatctatatt accctgttat ccctagcgga
     2101 tctgccggta gaggtgtggt caataagagc gacctcatac tatacctgag aaagcaacct
     2161 gacctacagg aaagagttac tcaagaataa gaattttcgt tttaaaacct aagagtcact
     2221 ttaaaatttg tatacactta ttttttttat aacttattta ataataaaaa tcataaatca
     2281 taagaaattc gcttatttag aagtggcgcg ccctacagga acaggtggtg gcggccctcg
     2341 gcgcgctcgt actgctccac gatggtgtag tcctcgttgt gggaggtgat gtccagcttg
     2401 gagtccacgt agtagtagcc gggcagctgc acgggcttct tggccatgta gatggacttg
     2461 aactccacca ggtagtggcc gccgtccttc agcttcaggg ccttgtggat ctcgcccttc
     2521 agcacgccgt cgcgggggta caggcgctcg gtggaggcct cccagcccat agtcttcttc
     2581 tgcattacgg ggccgtcgga ggggaagttc acgccgatga acttcacctt gtagatgaag
     2641 gagccgtcct gcagggagga gtcctgggtc acggtcacca cgccgccgtc ctcgaagttc
     2701 atcacgcgct cccacttgaa gccctcgggg aaggacagct tcttgtagtc ggggatgtcg
     2761 gcggggtgct tcacgtacac cttggagccg tactggaact ggggggacag gatgtcccag
     2821 gcgaagggca gggggccgcc cttggtcacc ttcagcttgg cggtctgggt gccctcgtag
     2881 gggcggccct cgccctcgcc ctcgatctcg aactcgtggc cgttcacgga gccctccatg
     2941 cgcaccttga agcgcatgaa ctccttgatg acgtcctcgg aggaggccat gttaattaat
     3001 gttaattcag taaattttcg atcttgggaa gaaaaaagca gtaagcgtga aaaatctaaa
     3061 agctgatgta gtagaagatc ctattcttta acaaagattg accttttctt tttcttcttg
     3121 gtttgagtag aaaggggaag gaagaataca agagagagga aaaaaaggaa gataaaaaga
     3181 gagcgtgata taaatgaata tatattaaac aagagagatt gggaaggaaa ggatcaaaca
     3241 aacccaaaaa tatttcaaaa aggagagaga gaggcgagtt tggtttcaaa acggtttatt
     3301 tatttatgca agaggacgtg gaagaaaaag aagaaggaag aaaaaaattt gaaagaaaaa
     3361 aacgcgtggc gggtaaagaa gaaaatggaa aatagaggcc gggtgacaga gaaatattga
     3421 gggttaattg gaaaatatgt tagggtgagg catatgtttt taagggtttt gaggatccac
     3481 tagttctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt
     3541 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3601 tcacaattcc acacaacata ggagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3661 gagtgaggta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3721 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3781 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3841 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3901 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3961 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4021 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4081 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4141 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4201 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4261 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4321 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4381 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     4441 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4501 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4561 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4621 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4681 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4741 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4801 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4861 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4921 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4981 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5041 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5101 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5161 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5221 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5281 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5341 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5401 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5461 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5521 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5581 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5641 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5701 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     5761 ataggcgtat cacgaggccc tttcgtc
//
