LOCUS       sequence-8469-ex        5677 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 5677)
  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 5677)
  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: pJB06T
FEATURES             Location/Qualifiers
     source          1..5677
                     /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        187..467
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             468..1142
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1241..1696)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1378..1399)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1590..1609
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1826..1848
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1840..1857
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1841..1857
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1867..1886
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1867..1885
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1910..1926
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1911..1927
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(1995..2182)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     2103..2127
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2203..2880)
                     /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(2265..2288)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2463..2482)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2680..2700
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2732..2750
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(3364..3380)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3364..3380)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3417..3437)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3417..3435)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3456..3472)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3456..3472)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3469..3491)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3480..3496
                     /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(3504..3510,3511..3528,3529..3534))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3687..3704)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3858..4446)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3938..3957)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4617..5408,5409..5477))
                     /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     5240..5259
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5478..5582)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5650..5668
                     /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 accataaacg acattactat atatataata taggaagcat ttaatagaca gcatcgtaat
      241 atatgtgtac tttgcagtta tgacgccaga tggcagtagt ggaagatatt ctttattgaa
      301 aaatagcttg tcaccttacg tacaatcttg atccggagct tttctttttt tgccgattaa
      361 gaattaattc ggtcgaaaaa agaaaaggag agggccaaga gggagggcat tggtgactat
      421 tgagcacgtg agtatacgtg attaagcaca caaaggcagc ttggagtatg tctgttatta
      481 atttcacagg tagttctggt ccattggtga aagtttgcgg cttgcagagc acagaggccg
      541 cagaatgtgc tctagattcc gatgctgact tgctgggtat tatatgtgtg cccaatagaa
      601 agagaacaat tgacccggtt attgcaagga aaatttcaag tcttgtaaaa gcatataaaa
      661 atagttcagg cactccgaaa tacttggttg gcgtgtttcg taatcaacct aaggaggatg
      721 ttttggctct ggtcaatgat tacggcattg atatcgtcca actgcatgga gatgagtcgt
      781 ggcaagaata ccaagagttc ctcggtttgc cagttattaa aagactcgta tttccaaaag
      841 actgcaacat actactcagt gcagcttcac agaaacctca ttcgtttatt cccttgtttg
      901 attcagaagc aggtgggaca ggtgaacttt tggattggaa ctcgatttct gactgggttg
      961 gaaggcaaga gagccccgaa agcttacatt ttatgttagc tggtggactg acgccagaaa
     1021 atgttggtga tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg
     1081 agacaaatgg tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat
     1141 aggttattac tgagtagtat ttatttaagt attgtttgtg cacttgccta tgcggtgtga
     1201 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt
     1261 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     1321 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     1381 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     1441 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     1501 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     1561 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     1621 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     1681 ccgctacagg gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     1741 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     1801 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
     1861 cgcgcgtaat acgactcact atagggcgaa ttgggtaccg ggccccccct cgaggtcgac
     1921 ggtatcgata agcttgatat cgaattcctg cagcccgggg gatctatatt accctgttat
     1981 ccctagcgga tctgccggta gaggtgtggt caataagagc gacctcatac tatacctgag
     2041 aaagcaacct gacctacagg aaagagttac tcaagaataa gaattttcgt tttaaaacct
     2101 aagagtcact ttaaaatttg tatacactta ttttttttat aacttattta ataataaaaa
     2161 tcataaatca taagaaattc gcttatttag aagtggcgcg ccctacagga acaggtggtg
     2221 gcggccctcg gcgcgctcgt actgctccac gatggtgtag tcctcgttgt gggaggtgat
     2281 gtccagcttg gagtccacgt agtagtagcc gggcagctgc acgggcttct tggccatgta
     2341 gatggacttg aactccacca ggtagtggcc gccgtccttc agcttcaggg ccttgtggat
     2401 ctcgcccttc agcacgccgt cgcgggggta caggcgctcg gtggaggcct cccagcccat
     2461 agtcttcttc tgcattacgg ggccgtcgga ggggaagttc acgccgatga acttcacctt
     2521 gtagatgaag gagccgtcct gcagggagga gtcctgggtc acggtcacca cgccgccgtc
     2581 ctcgaagttc atcacgcgct cccacttgaa gccctcgggg aaggacagct tcttgtagtc
     2641 ggggatgtcg gcggggtgct tcacgtacac cttggagccg tactggaact ggggggacag
     2701 gatgtcccag gcgaagggca gggggccgcc cttggtcacc ttcagcttgg cggtctgggt
     2761 gccctcgtag gggcggccct cgccctcgcc ctcgatctcg aactcgtggc cgttcacgga
     2821 gccctccatg cgcaccttga agcgcatgaa ctccttgatg acgtcctcgg aggaggccat
     2881 gttaattaat gttaattcag taaattttcg atcttgggaa gaaaaaagca gtaagcgtga
     2941 aaaatctaaa agctgatgta gtagaagatc ctattcttta acaaagattg accttttctt
     3001 tttcttcttg gtttgagtag aaaggggaag gaagaataca agagagagga aaaaaaggaa
     3061 gataaaaaga gagcgtgata taaatgaata tatattaaac aagagagatt gggaaggaaa
     3121 ggatcaaaca aacccaaaaa tatttcaaaa aggagagaga gaggcgagtt tggtttcaaa
     3181 acggtttatt tatttatgca agaggacgtg gaagaaaaag aagaaggaag aaaaaaattt
     3241 gaaagaaaaa aacgcgtggc gggtaaagaa gaaaatggaa aatagaggcc gggtgacaga
     3301 gaaatattga gggttaattg gaaaatatgt tagggtgagg catatgtttt taagggtttt
     3361 gaggatccac tagttctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt
     3421 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     3481 tgttatccgc tcacaattcc acacaacata ggagccggaa gcataaagtg taaagcctgg
     3541 ggtgcctaat gagtgaggta actcacatta attgcgttgc gctcactgcc cgctttccag
     3601 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3661 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3721 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3781 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3841 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3901 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3961 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4021 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4081 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4141 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4201 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4261 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     4321 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4381 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4441 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4501 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4561 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4621 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4681 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4741 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4801 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4861 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4921 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4981 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5041 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5101 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5161 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5221 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5281 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5341 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5401 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5461 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5521 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5581 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     5641 acctataaaa ataggcgtat cacgaggccc tttcgtc
//
