LOCUS       sequence-13252-e        6395 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 6395)
  AUTHORS   Tan C, Marguet P, You L
  TITLE     Emergent bistability by a growth-modulating positive feedback 
            circuit.
  JOURNAL   Nat Chem Biol. 2009 Nov . 5(11):842-8.
  PUBMED    19801994
REFERENCE   2  (bases 1 to 6395)
  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: "Nat Chem 
            Biol. 2009 Nov . 5(11):842-8."
COMMENT     Sequence Label: pCFP
FEATURES             Location/Qualifiers
     source          1..6395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     178..197
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     281..300
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        281..299
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    300..324
                     /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)."
     RBS             336..347
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             356..1069
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKA
                     HFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(386..414)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1009..1029
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     RBS             1098..1109
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(1173..1191)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1187..1234
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1409..1414,1415..1431,1432..1437))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1464..1482)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1550..1654
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1655..1723,1724..2515)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1873..1892)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2686..3274
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3175..3194
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3428..3445
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3460..3602
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3528..3547)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3688..3710
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3704..3895)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5207..6289)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6251..6270
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6290..6367)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag
       61 gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat
      121 acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt
      181 gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat
      241 gcgtccggcg tagaggatcg agatctcgat cccgcgaaat taatacgact cactataggg
      301 gaattgtgag cggataacaa ttcccctcta gaattaaaga ggagaaagct accgcatgcg
      361 taaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt
      421 taatgggcac aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact
      481 tacccttaaa tttatttgca ctactggaaa actacctgtt ccatggccaa cacttgtcac
      541 tactttgact tggggtgttc aatgcttttc aagataccca gatcatatga aacggcatga
      601 ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa agaactatat ttttcaaaga
      661 tgacgggaac tacaagacac gtgctgaagt caagtttgaa ggtgataccc ttgttaatag
      721 aatcgagtta aaaggtattg attttaaaga agatggaaac attcttggac acaaattgga
      781 atacaactat atttcacaca atgtatacat cactgcagac aaacaaaaga atggaatcaa
      841 agctcatttc aaaattagac acaacattga agatggaagc gttcaactag cagaccatta
      901 tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc
      961 cacacaatct gccctttcga aagatcccaa cgaaaagaga gaccacatgg tccttcttga
     1021 gtttgtaaca gctgctggga ttacacatgg catggatgaa ctatacaaat aagagctcca
     1081 ttagcatcgc tagcattaaa gaggagaaag ctaccgcctc gaggatccgg ctgctaacaa
     1141 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     1201 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata
     1261 tcccgcaaga ggcccggcag taccggcata accaagccta tgcctacagc atccagggtg
     1321 acggtgccga ggatgacgat gagcgcattg ttagatttca tacacggtgc ctgactgcgt
     1381 tagcaattta actgtgataa actaccgcat taaagcttat cgatgataag ctgtcaaaca
     1441 tgagaattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     1501 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     1561 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     1621 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     1681 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     1741 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     1801 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     1861 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     1921 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     1981 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     2041 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     2101 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     2161 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     2221 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     2281 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     2341 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     2401 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     2461 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     2521 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     2581 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     2641 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     2701 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2761 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2821 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     2881 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2941 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3001 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3061 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3121 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3181 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3241 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3301 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     3361 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     3421 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     3481 tacgcatctg tgcggtattt cacaccgcat atatggtgca ctctcagtac aatctgctct
     3541 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     3601 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3661 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3721 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     3781 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     3841 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     3901 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     3961 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     4021 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     4081 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     4141 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     4201 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     4261 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     4321 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc
     4381 ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag
     4441 ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg
     4501 tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc
     4561 accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa
     4621 cccgttccat gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccagtgat
     4681 cgaagttagg ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc
     4741 tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag
     4801 aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag
     4861 cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
     4921 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
     4981 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
     5041 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
     5101 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
     5161 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     5221 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5281 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
     5341 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
     5401 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
     5461 cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa
     5521 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
     5581 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
     5641 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
     5701 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
     5761 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
     5821 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
     5881 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
     5941 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
     6001 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
     6061 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
     6121 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
     6181 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
     6241 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     6301 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
     6361 tggtgtccgg gatctcgacg ctctccctta tgcga
//
