LOCUS       sequence-9993-ex        6797 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 6797)
  AUTHORS   Yu RC, Pesce CG, Colman-Lerner A, Lok L, Pincus D, Serra E, Holl M, 
            Benjamin K, Gordon A, Brent R
  TITLE     Negative feedback that improves information transmission in yeast 
            signalling.
  JOURNAL   Nature. 2008 Dec 11. 456(7223):755-61.
  PUBMED    19079053
REFERENCE   2  (bases 1 to 6797)
  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. 
            2008 Dec 11. 456(7223):755-61."
COMMENT     Sequence Label: pES1002
FEATURES             Location/Qualifiers
     source          1..6797
                     /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     complement(4509..4529)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4509..4527)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4548..4564)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4548..4564)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4561..4583)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4572..4588
                     /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(4596..4602,4603..4620,4621..4626))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4779..4796)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4950..5538)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5030..5049)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5709..6500,6501..6569))
                     /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     6332..6351
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6570..6674)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6770..6788
                     /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 ttgggtacca atcatcaaag cttaatatat caaatgaatc
     2041 ttgcgcgata gtctttgtca ctttgggggt gaattcttcg gcattagtta aaggaatgag
     2101 aagtaaggcc caaagagaat agacaaaatg aagtaatatc ataacgtgtg atgactactg
     2161 gacacttcag ggcttttcca gatatacaga gctttataat acaatctaag ggcaaaataa
     2221 aaaaggaaaa agagtacggt agttgagaaa aaaaccggaa aggaattgct ttaaagatct
     2281 tagtagaatg aaatagtacc acctactcgc ctcctttttt tgtgcttctt gagaggtctt
     2341 ttgagctgtt gcatcatgct gttatttaga ggttattttt tgaccttggt ttttccttct
     2401 tttcgccgta ctcagcccga gaaaaaagca gcgactgcac taaactagag gaaagaaggc
     2461 ttaaaacatc acgcgatcca tataaagacc cgagtagttg gagagattga ctacttgacg
     2521 ttcaaaagaa catacataag gataaattcg ttcggacttt ttaacttaca cacatctttt
     2581 ggtgttcatt caaggggcgg tagtgcttgt agtccgcaca ggcggatact ttttagtttg
     2641 ggaagagtta actggagagg gcagtttact ttcacatact cttttatctt tctgaagtaa
     2701 aatgtgatgt acctacaagc aatgcataag ctcttgagtt ttagatctgt ttcaccctgg
     2761 aactcaaaat tcttttactc gaaattttta cttttttttt ttttgtttct gcattctctc
     2821 agattttaga tgatgcggtt ttttacaggg cattgaaaca attgcagaaa agcaacatac
     2881 taatatatca taacttttta ctcttgcctc tcagaaacta tatatacgtt gtaatcattt
     2941 tctttcttct aatagctagt tcgtttgaac tacaaggaaa taaggcagag aaaaagaaag
     3001 gaaaataata tgccaaagag aattgtatac aatatatcca gtgacttcca gttgaagtcg
     3061 ttactgggag agggtgcata cggtgtggta tgttctgcaa cgcataagcc cacgggagaa
     3121 atcgtggcaa taaaaaagat cgaaccattc gataagcctt tgttcgcatt acgtacgctg
     3181 cgtgaaataa agatcctgaa gcacttcaag cacgaaaata tcataacaat cttcaacatt
     3241 caacgccctg actcgttcga aaacttcaat gaggtctaca taattgctga gctaatgcag
     3301 acagatttac accgtgtaat ctccacccag atgctgagtg acgatcatat acaatatttt
     3361 atataccaaa ccttgagagc agtgaaagtg ctgcatggtt cgaacgtcat ccatcgtgat
     3421 ttaaagccct ccaaccttct cataaactcc aactgtgact tgaaagtatg tgatttcggt
     3481 ttagcaagaa tcattgacga gtcagccgcg gacaattcag agcccacagg tcagcaaagc
     3541 ggcatgaccg agtatgtggc cacacgttgg tacagggcgc cagaggtgat gttaacctct
     3601 gccaaatact caagggccat ggacgtgtgg tcctgcggat gtattctcgc tgaacttttc
     3661 ttaagacggc caatcttccc tggcagagat tatcgccatc aactactact gatattcggt
     3721 atcatcggta cacctcactc agataatgat ttgcggtgta tagagtcacc cagggctaga
     3781 gagtacataa agtcgcttcc catgtaccct gccgcgccac tggagaagat gttccctcga
     3841 gtcaacccga aaggcataga tcttttacag cgtatgcttg tttttgaccc tgcgaagagg
     3901 attactgcta aggaggcact ggagcatccg tatttgcaaa cataccacga tccaaacgac
     3961 gaacctgaag gcgaacccat cccacccagc ttcttcgagt ttgatcacta caaggaggca
     4021 ctaacgacga aagacctcaa gaaactcatt tggaacgaaa tatttagtta gccatcatta
     4081 tcattaaaat atcaacccga agaacaatgt atacatatac atatacgtac acatatacat
     4141 atgtacatat gacatacgta ttagccgctg aggacgcgga cgtataaaag gacaatactt
     4201 atatggagct aaggggagca gttacgcaac tccgtgatcg cgcgccacgg gccgtcggcg
     4261 gctgttaatt gaagaaaaaa aaaatgaaga accacaaggg gtgatccata taggtgacta
     4321 gcatcatccc ctgcgacgcg cggcccgccg ggccaaaggc gggcaatgcg cgctgctgat
     4381 tggcctcgag gacaacgccc tcaaccacat ccgcaacagc caatcccatc ggagcgtcaa
     4441 actaccaaag tagtgattgt atggatgacc acgcggccgc caccgcggtg gagctccagc
     4501 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     4561 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca taggagccgg aagcataaag
     4621 tgtaaagcct ggggtgccta atgagtgagg taactcacat taattgcgtt gcgctcactg
     4681 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4741 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4801 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4861 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4921 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4981 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5041 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5101 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5161 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5221 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5281 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5341 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5401 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5461 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5521 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5581 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5641 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5701 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5761 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5821 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5881 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5941 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6001 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6061 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6121 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6181 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6241 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6301 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6361 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6421 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6481 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6541 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6601 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6661 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ggtccttttc gacgtcaggc
     6721 aagcgatccg tcctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     6781 cacgaggccc tttcgtc
//
