LOCUS       sequence-13253-e        9046 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 9046)
  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 9046)
  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: pCFPT7
FEATURES             Location/Qualifiers
     source          1..9046
                     /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(3824..3842)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3838..3885
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4060..4065,4066..4082,4083..4088))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(4115..4133)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4201..4305
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4306..4374,4375..5166)
                     /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(4524..4543)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5337..5925
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5826..5845
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6079..6096
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6111..6253
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6179..6198)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6339..6361
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6355..6546)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(7858..8940)
                     /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     8902..8921
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8941..9018)
                     /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 ctaccgcatg aacacgatta acatcgctaa
     1141 gaacgacttc tctgacatcg aactggctgc tatcccgttc aacactctgg ctgaccatta
     1201 cggtgagcgt ttagctcgcg aacagttggc ccttgagcat gagtcttacg agatgggtga
     1261 agcacgcttc cgcaagatgt ttgagcgtca acttaaagct gtgaggttgc ggataacgct
     1321 gccgccaagc ctctcatcac taccctactc cctaagatga ttgcacgcat caacgactgg
     1381 tttgaggaag tgaaagctaa gcgcggcaag cgcccgacag ccttccagtt cctgcaagaa
     1441 atcaagccgg aagccgtagc gtacatcacc attaagacca ctctggcttg cctaaccagt
     1501 gctgacaata caaccgttca ggctgtagca agcgcaatcg gtcgggccat tgaggacgag
     1561 gctcgcttcg gtcgtatccg tgaccttgaa gctaagcact tcaagaaaaa cgttgaggaa
     1621 caactcaaca agcgcgtagg gcacgtctac aagaaagcat ttatgcaagt tgtcgaggct
     1681 gacatgctct ctaagggtct actcggtggc gaggcgtggt cttcgtggca taaggaagac
     1741 tctattcatg taggagtacg ctgcatcgag atgctcattg agtcaaccgg aatggttagc
     1801 ttacaccgcc aaaatgctgg cgtagtaggt caagactctg agactatcga actcgcacct
     1861 gaatacgctg aggctatcgc aacccgtgca ggtgcgctgg ctggcatctc tccgatgttc
     1921 caaccttgcg tagttcctcc taagccgtgg actggcatta ctggtggtgg ctattgggct
     1981 aacggtcgtc gtcctctggc gctggtgcgt actcacagta agaaagcact gatgcgctac
     2041 gaagacgttt acatgcctga ggtgtacaaa gcgattaaca ttgcgcaaaa caccgcatgg
     2101 aaaatcaaca agaaagtcct agcggtcgcc aacgtaatca ccaagtggaa gcattgtccg
     2161 gtcgaggaca tccctgcgat tgagcgtgaa gaactcccga tgaaaccgga agacatcgac
     2221 atgaatcctg aggctctcac cgcgtggaaa cgtgctgccg ctgctgtgta ccgcaaggac
     2281 aaggctcgca agtctcgccg tatcagcctt gagttcatgc ttgagcaagc caataagttt
     2341 gctaaccata aggccatctg gttcccttac aacatggact ggcgcggtcg tgtttacgct
     2401 gtgtcaatgt tcaacccgca aggtaacgat atgaccaaag gactgcttac gctggcgaaa
     2461 ggtaaaccaa tcggtaagga aggttactac tggctgaaaa tccacggtgc aaactgtgcg
     2521 ggtgtcgata aggttccgtt ccctgagcgc atcaagttca ttgaggaaaa ccacgagaac
     2581 atcatggctt gcgctaagtc tccactggag aacacttggt gggctgagca agattctccg
     2641 ttctgcttcc ttgcgttctg ctttgagtac gctggggtac agcaccacgg cctgagctat
     2701 aactgctccc ttccgctggc gtttgacggg tcttgctctg gcatccagca cttctccgcg
     2761 atgctccgag atgaggtagg tggtcgcgcg gttaacttgc ttcctagtga aaccgttcag
     2821 gacatctacg ggattgttgc taagaaagtc aacgagattc tacaagcaga cgcaatcaat
     2881 gggaccgata acgaagtagt taccgtgacc gatgagaaca ctggtgaaat ctctgagaaa
     2941 gtcaagctgg gcactaaggc actggctggt caatggctgg cttacggtgt tactcgcagt
     3001 gtgactaagc gttcagtcat gacgctggct tacgggtcca aagagttcgg cttccgtcaa
     3061 caagtgctgg aagataccat tcagccagct attgattccg gcaagggtct gatgttcact
     3121 cagccgaatc aggctgctgg atacatggct aagctgattt gggaatctgt gagcgtgacg
     3181 gtggtagctg cggttgaagc aatgaactgg cttaagtctg ctgctaagct gctggctgct
     3241 gaggtcaaag ataagaagac tggagagatt cttcgcaagc gttgcgctgt gcattgggta
     3301 actcctgatg gtttccctgt gtggcaggaa tacaagaagc ctattcagac gcgcttgaac
     3361 ctgatgttcc tcggtcagtt ccgcttacag cctaccatta acaccaacaa agatagcgag
     3421 attgatgcac acaaacagga gtctggtatc gctcctaact ttgtacacag ccaagacggt
     3481 agccaccttc gtaagactgt agtgtgggca cacgagaagt acggaatcga atcttttgca
     3541 ctgattcacg actccttcgg taccattccg gctgacgctg cgaacctgtt caaagcagtg
     3601 cgcgaaacta tggttgacac atatgagtct tgtgatgtac tggctgattt ctacgaccag
     3661 ttcgctgacc agttgcacga gtctcaattg gacaaaatgc cagcacttcc ggctaaaggt
     3721 aacttgaacc tccgtgacat cttagagtcg gacttcgcgt tcgcgtaact cgaggatccg
     3781 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     3841 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
     3901 atatccggat atcccgcaag aggcccggca gtaccggcat aaccaagcct atgcctacag
     3961 catccagggt gacggtgccg aggatgacga tgagcgcatt gttagatttc atacacggtg
     4021 cctgactgcg ttagcaattt aactgtgata aactaccgca ttaaagctta tcgatgataa
     4081 gctgtcaaac atgagaattc ttgaagacga aagggcctcg tgatacgcct atttttatag
     4141 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     4201 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4261 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4321 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4381 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4441 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4501 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     4561 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4621 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4681 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4741 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4801 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     4861 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4921 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4981 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5041 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5101 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5161 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5221 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5281 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5341 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5401 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5461 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     5521 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5581 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5641 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5701 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5761 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5821 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5881 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5941 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6001 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6061 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     6121 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta
     6181 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     6241 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     6301 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     6361 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     6421 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     6481 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     6541 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     6601 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     6661 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     6721 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     6781 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     6841 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     6901 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     6961 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga
     7021 cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat
     7081 gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat
     7141 tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc
     7201 cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca
     7261 atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc
     7321 ggtccagtga tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga
     7381 tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg
     7441 gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag
     7501 acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     7561 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     7621 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
     7681 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
     7741 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     7801 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     7861 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     7921 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     7981 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     8041 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     8101 tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
     8161 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     8221 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     8281 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     8341 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     8401 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
     8461 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     8521 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     8581 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     8641 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     8701 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     8761 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     8821 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     8881 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     8941 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     9001 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcga
//
