LOCUS       sequence-9994-ex        5820 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 5820)
  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 5820)
  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: pES1004
FEATURES             Location/Qualifiers
     source          1..5820
                     /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(3532..3552)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3532..3550)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3571..3587)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3571..3587)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3584..3606)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3595..3611
                     /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(3619..3625,3626..3643,3644..3649))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3802..3819)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3973..4561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4053..4072)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4732..5523,5524..5592))
                     /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     5355..5374
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5593..5697)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5793..5811
                     /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 atatcccttc ccaatataaa ctcgtagatt
     2041 taataggtga gggagcgtac ggaacagtat gttcagcaat tcataagcct tccggcataa
     2101 aggtagctat caagaaaata caaccgttta gcaaaaaatt gtttgttaca agaactatac
     2161 gtgagatcaa gcttttacgg tatttccatg aacacgaaaa cataataagt atattggata
     2221 aagtaaggcc agtatccata gacaaactaa acgctgttta tttagtcgct gagttgatgg
     2281 aaaccgattt acaaaaagta attaataatc agaatagcgg gttttccact ttaagtgatg
     2341 accatgttca atactttaca taccaaatcc tcagagcctt aaagtctatt cacagtgcac
     2401 aagttatcca tagagacata aagccatcaa acctgttact aaattccaat tgtgatctca
     2461 aagtctgcga ttttggacta gcgaggtgtt tagctagcag tagcgattca agagaaacat
     2521 tggtaggatt catgacggag tacgtcgcaa cgcgatggta cagggcaccc gagataatgc
     2581 taacttttca agagtacaca actgcgatgg atatatggtc atgcggatgc attttggctg
     2641 aaatggtctc cgggaagcct ttgttcccag gcagagacta tcatcatcaa ttatggctaa
     2701 ttctagaagt cttgggaact ccatctttcg aagactttaa tcagatcaaa tccaagaggg
     2761 ctaaagagta tatagcaaac ttacctatga ggccaccctt gccatgggag accgtctggt
     2821 caaagaccga tctgaatcca gatatgatag atttactaga caaaatgctt caattcaatc
     2881 ctgacaaaag aataagcgca gcagaagctt taagacaccc ttacctggca atgtaccatg
     2941 acccaagtga tgagccggaa tatcctccac ttaatttgga tgatgaattt tggaaactgg
     3001 ataacaagat aatgcgtccg gaagaggagg aagaagtgcc catagaaatg ctcaaagaca
     3061 tgctttacga tgaactaatg aagaccatgg aatagtattc acaagaacat ttctgccata
     3121 cttctaaaat ttccctatat tcagcttagc agtgacacgt tgtggtctgt aggtcaatat
     3181 gtaagtaaga aacttcaact cacatatgca cgatgcatgc caatggaaaa atgcaaggaa
     3241 cgaaatggcg ccacggcaac aagttttttt ttttcgccag cagaagtaca cgaaatgcgg
     3301 cttcatgagc ctcttcactg ctttgcctaa acgggaaatg cagagaaaaa ccagccatcg
     3361 cgtgtgcttg gagagctgac gcgactgtaa tcaaagaggc gatatcaaca ccttttatcc
     3421 agcactattc aacagtgaat gggctcccaa gtaagtcttg gcattgtgct ttctattctt
     3481 aagtattaag tagaagcggc cgccaccgcg gtggagctcc agcttttgtt ccctttagtg
     3541 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3601 tccgctcaca attccacaca acataggagc cggaagcata aagtgtaaag cctggggtgc
     3661 ctaatgagtg aggtaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3721 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3781 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3841 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3901 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3961 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4021 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4081 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4141 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4201 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4261 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4321 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4381 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     4441 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4501 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4561 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4621 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4681 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4741 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4801 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4861 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4921 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4981 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5041 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5101 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5161 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5221 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5281 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5341 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5401 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5461 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5521 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5581 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5641 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5701 atttccccga aaagtgccac ctgggtcctt ttcgacgtca ggcaagcgat ccgtcctaag
     5761 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
