LOCUS       sequence-178135-        7925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic gene circuit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7925)
  AUTHORS   Rubens JR, Selvaggio G, Lu TK
  TITLE     Synthetic mixed-signal computation in living cells.
  JOURNAL   Nat Commun. 2016 Jun 3;7:11658. doi: 10.1038/ncomms11658.
  PUBMED    27255669
REFERENCE   2  (bases 1 to 7925)
  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 
            Commun."; date: "2016-06-3"; volume: "7:11658. doi"; pages: " 
            10.1038/ncomms11658"
COMMENT     Sequence Label: pBAC Bxbi TetR
FEATURES             Location/Qualifiers
     source          1..7925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        1..50
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             93..101
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             107..730
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(180..199)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      790..876
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    896..944
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     primer_bind     complement(1117..1135)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(1178..1837)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1752..1771
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1838..1940)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      2798..3412
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      2889..3420
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      3488..3707
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             3798..4553
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    4556..4806
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             5132..6307
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             6307..7278
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     protein_bind    7860..7917
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attP site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
ORIGIN
        1 ctgtcaccgg atgtgctttc cggtctgatg agtccgtgag gacgaaacag cctctacaaa
       61 taattttgtt taagcgttag cggaaataag attaaggagg ttcaacatgt ctagattaga
      121 taaaagtaaa gtgattaaca gcgcattaga gctgcttaat gaggtcggaa tcgaaggttt
      181 aacaacccgt aaactcgccc agaagctagg tgtagagcag cctacattgt attggcatgt
      241 aaaaaataag cgggctttgc tcgacgcctt agccattgag atgttagata ggcaccatac
      301 tcacttttgc cctttagaag gggaaagctg gcaagatttt ttacgtaata acgctaaaag
      361 ttttagatgt gctttactaa gtcatcgcga tggagcaaaa gtacatttag gtacacggcc
      421 tacagaaaaa cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc aacaaggttt
      481 ttcactagag aatgcattat atgcactcag cgctgtgggg cattttactt taggttgcgt
      541 attggaagat caagagcatc aagtcgctaa agaagaaagg gaaacaccta ctactgatag
      601 tatgccgcca ttattacgac aagctatcga attatttgat caccaaggtg cagagccagc
      661 cttcttattc ggccttgaat tgatcatatg cggattagaa aaacaactta aatgtgaaag
      721 tgggtcttaa aagcttgata tcgaattcct gcagcccggg ggatcccatg gtacgcgtgc
      781 tagaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
      841 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgccctagaa tcgatgcccg
      901 gatgatcctg acgacggaga ccgccgtcgt cgacaagccg gccgttaatt aaaaagttaa
      961 acaaaattat ttgtagaggg aaaccgttgt ggtctccctg aatatattat acgagcctta
     1021 tgcatgcccg taaagttatc cagcaaccac tcatagacct agggcagcag atagggacga
     1081 cgtggtgtta gctgtgctcg aggtgaagac gaaagggcct cgtgatacgc ctatttttat
     1141 aggttaatgt catgataata atggtttctt agacgtctta cgccccgccc tgccactcat
     1201 cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat
     1261 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
     1321 tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga
     1381 aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat
     1441 aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga
     1501 aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg
     1561 tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgaa
     1621 attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt
     1681 gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat
     1741 aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata
     1801 tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa
     1861 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
     1921 aacctcttac gtgccgatca acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg
     1981 tatcaacagg gacaccagga tttatttatt ctgcgaagtg atcttccgtc acaggtattt
     2041 attcgcgata agctcatgga gcggcgtaac cgtcgcacag gaaggacaga gaaagcgcgg
     2101 atctgggaag tgacggacag aacggtcagg acctggattg gggaggcggt tgccgccgct
     2161 gctgctgacg gtgtgacgtt ctctgttccg gtcacaccac atacgttccg ccattcctat
     2221 gcgatgcaca tgctgtatgc cggtataccg ctgaaagttc tgcaaagcct gatgggacat
     2281 aagtccatca gttcaacgga agtctacacg aaggtttttg cgctggatgt ggctgcccgg
     2341 caccgggtgc agtttgcgat gccggagtct gatgcggttg cgatgctgaa acaattatcc
     2401 tgagaataaa tgccttggcc tttatatgga aatgtggaac tgagtggata tgctgttttt
     2461 gtctgttaaa cagagaagct ggctgttatc cactgagaag cgaacgaaac agtcgggaaa
     2521 atctcccatt atcgtagaga tccgcattat taatctcagg agcctgtgta gcgtttatag
     2581 gaagtagtgt tctgtcatga tgcctgcaag cggtaacgaa aacgatttga atatgccttc
     2641 aggaacaata gaaatcttcg tgcggtgtta cgttgaagtg gagcggatta tgtcagcaat
     2701 ggacagaaca acctaatgaa cacagaacca tgatgtggtc tgtcctttta cagccagtag
     2761 tgctcgccgc agtcgagcga cagggcgaag ccctcggctg gttgccctcg ccgctgggct
     2821 ggcggccgtc tatggccctg caaacgcgcc agaaacgccg tcgaagccgt gtgcgagaca
     2881 ccgcggccgg ccgccggcgt tgtggatacc tcgcggaaaa cttggccctc actgacagat
     2941 gaggggcgga cgttgacact tgaggggccg actcacccgg cgcggcgttg acagatgagg
     3001 ggcaggctcg atttcggccg gcgacgtgga gctggccagc ctcgcaaatc ggcgaaaacg
     3061 cctgatttta cgcgagtttc ccacagatga tgtggacaag cctggggata agtgccctgc
     3121 ggtattgaca cttgaggggc gcgactactg acagatgagg ggcgcgatcc ttgacacttg
     3181 aggggcagag tgctgacaga tgaggggcgc acctattgac atttgagggg ctgtccacag
     3241 gcagaaaatc cagcatttgc aagggtttcc gcccgttttt cggccaccgc taacctgtct
     3301 tttaacctgc ttttaaacca atatttataa accttgtttt taaccagggc tgcgccctgt
     3361 gcgcgtgacc gcgcacgccg aaggggggtg cccccccttc tcgaaccctc ccggtcgagt
     3421 gagcgaggaa gcaccaggga acagcactta tatattctgc ttacacacga tgcctgaaaa
     3481 aacttccctt ggggttatcc acttatccac ggggatattt ttataattat tttttttata
     3541 gtttttagat cttctttttt agagcgcctt gtaggccttt atccatgctg gttctagaga
     3601 aggtgttgtg acaaattgcc ctttcagtgt gacaaatcac cctcaaatga cagtcctgtc
     3661 tgtgacaaat tgcccttaac cctgtgacaa attgccctca gaagaagctg ttttttcaca
     3721 aagttatccc tgcttattga ctctttttta tttagtgtga caatctaaaa acttgtcaca
     3781 cttcacatgg atctgtcatg gcggaaacag cggttatcaa tcacaagaaa cgtaaaaata
     3841 gcccgcgaat cgtccagtca aacgacctca ctgaggcggc atatagtctc tcccgggatc
     3901 aaaaacgtat gctgtatctg ttcgttgacc agatcagaaa atctgatggc accctacagg
     3961 aacatgacgg tatctgcgag atccatgttg ctaaatatgc tgaaatattc ggattgacct
     4021 ctgcggaagc cagtaaggat atacggcagg cattgaagag tttcgcgggg aaggaagtgg
     4081 ttttttatcg ccctgaagag gatgccggcg atgaaaaagg ctatgaatct tttccttggt
     4141 ttatcaaacg tgcgcacagt ccatccagag ggctttacag tgtacatatc aacccatatc
     4201 tcattccctt ctttatcggg ttacagaacc ggtttacgca gtttcggctt agtgaaacaa
     4261 aagaaatcac caatccgtat gccatgcgtt tatacgaatc cctgtgtcag tatcgtaagc
     4321 cggatggctc aggcatcgtc tctctgaaaa tcgactggat catagagcgt taccagctgc
     4381 ctcaaagtta ccagcgtatg cctgacttcc gccgccgctt cctgcaggtc tgtgttaatg
     4441 agatcaacag cagaactcca atgcgcctct catacattga gaaaaagaaa ggccgccaga
     4501 cgactcatat cgtattttcc ttccgcgata tcacttccat gacgacagga tagtctgagg
     4561 gttatctgtc acagatttga gggtggttcg tcacatttgt tctgacctac tgagggtaat
     4621 ttgtcacagt tttgctgttt ccttcagcct gcatggattt tctcatactt tttgaactgt
     4681 aatttttaag gaagccaaat ttgagggcag tttgtcacag ttgatttcct tctctttccc
     4741 ttcgtcatgt gacttgatat cgggggttag ttcgtcatca ttgatgaggg ttgattatca
     4801 cagtttatta ctctgaattg gctatccgcg tgtgtacctc tacctggagt ttttcccacg
     4861 gtggatattt cttcttgcgc tgagcgtaag agctatctga cagaacagtt cttctttgct
     4921 tcctcgccag ttcgctcgct atgctcggtt acacggctgc ggcgagcgct agtgataata
     4981 agtgactgag gtatgtgctc ttcttatctc cttttgtagt gttgctctta ttttaaacaa
     5041 ctttgcggtt ttttgatgac tttgcgattt tgttgttgct ttgcagtaaa ttgcaagatt
     5101 taataaaaaa acgcaaagca atgattaaag gatgttcaga atgaaactca tggaaacact
     5161 taaccagtgc ataaacgctg gtcatgaaat gacgaaggct atcgccattg cacagtttaa
     5221 tgatgacagc ccggaagcga ggaaaataac ccggcgctgg agaataggtg aagcagcgga
     5281 tttagttggg gtttcttctc aggctatcag agatgccgag aaagcagggc gactaccgca
     5341 cccggatatg gaaattcgag gacgggttga gcaacgtgtt ggttatacaa ttgaacaaat
     5401 taatcatatg cgtgatgtgt ttggtacgcg attgcgacgt gctgaagacg tatttccacc
     5461 ggtgatcggg gttgctgccc ataaaggtgg cgtttacaaa acctcagttt ctgttcatct
     5521 tgctcaggat ctggctctga aggggctacg tgttttgctc gtggaaggta acgaccccca
     5581 gggaacagcc tcaatgtatc acggatgggt accagatctt catattcatg cagaagacac
     5641 tctcctgcct ttctatcttg gggaaaagga cgatgtcact tatgcaataa agcccacttg
     5701 ctggccgggg cttgacatta ttccttcctg tctggctctg caccgtattg aaactgagtt
     5761 aatgggcaaa tttgatgaag gtaaactgcc caccgatcca cacctgatgc tccgactggc
     5821 cattgaaact gttgctcatg actatgatgt catagttatt gacagcgcgc ctaacctggg
     5881 tatcggcacg attaatgtcg tatgtgctgc tgatgtgctg attgttccca cgcctgctga
     5941 gttgtttgac tacacctccg cactgcagtt tttcgatatg cttcgtgatc tgctcaagaa
     6001 cgttgatctt aaagggttcg agcctgatgt acgtattttg cttaccaaat acagcaatag
     6061 taatggctct cagtccccgt ggatggagga gcaaattcgg gatgcctggg gaagcatggt
     6121 tctaaaaaat gttgtacgtg aaacggatga agttggtaaa ggtcagatcc ggatgagaac
     6181 tgtttttgaa caggccattg atcaacgctc ttcaactggt gcctggagaa atgctctttc
     6241 tatttgggaa cctgtctgca atgaaatttt cgatcgtctg attaaaccac gctgggagat
     6301 tagataatga agcgtgcgcc tgttattcca aaacatacgc tcaatactca accggttgaa
     6361 gatacttcgt tatcgacacc agctgccccg atggtggatt cgttaattgc gcgcgtagga
     6421 gtaatggctc gcggtaatgc cattactttg cctgtatgtg gtcgggatgt gaagtttact
     6481 cttgaagtgc tccggggtga tagtgttgag aagacctctc gggtatggtc aggtaatgaa
     6541 cgtgaccagg agctgcttac tgaggacgca ctggatgatc tcatcccttc ttttctactg
     6601 actggtcaac agacaccggc gttcggtcga agagtatctg gtgtcataga aattgccgat
     6661 gggagtcgcc gtcgtaaagc tgctgcactt accgaaagtg attatcgtgt tctggttggc
     6721 gagctggatg atgagcagat ggctgcatta tccagattgg gtaacgatta tcgcccaaca
     6781 agtgcttatg aacgtggtca gcgttatgca agccgattgc agaatgaatt tgctggaaat
     6841 atttctgcgc tggctgatgc ggaaaatatt tcacgtaaga ttattacccg ctgtatcaac
     6901 accgccaaat tgcctaaatc agttgttgct cttttttctc accccggtga actatctgcc
     6961 cggtcaggtg atgcacttca aaaagccttt acagataaag aggaattact taagcagcag
     7021 gcatctaacc ttcatgagca gaaaaaagct ggggtgatat ttgaagctga agaagttatc
     7081 actcttttaa cttctgtgct taaaacgtca tctgcatcaa gaactagttt aagctcacga
     7141 catcagtttg ctcctggagc gacagtattg tataagggcg ataaaatggt gcttaacctg
     7201 gacaggtccc gtgttccaac tgagtgtata gagaaaattg aggccattct taaggaactt
     7261 gaaaagccag caccctgatg cgaccacgtt ttagtctacg tttatctgtc tttacttaat
     7321 gtcctttgtt acaggccaga aagcataact ggcctgaata ttctctctgg gcccactgtt
     7381 ccacttgtat cgtcggtctg ataatcagac tgggaccacg gtcccactcg tatcgtcggt
     7441 ctgattatta gtctgggacc acggtcccac tcgtatcgtc ggtctgatta ttagtctggg
     7501 accacggtcc cactcgtatc gtcggtctga taatcagact gggaccacgg tcccactcgt
     7561 atcgtcggtc tgattattag tctgggacca tggtcccact cgtatcgtcg gtctgattat
     7621 tagtctggga ccacggtccc actcgtatcg tcggtctgat tattagtctg ggaccacgat
     7681 cccactcgtg ttgtcggtct gattatcggt ctgggaccac ggtcccactt gtattgtcga
     7741 tcagactatc agcgtgagac tacgattcca tcaatgcctg tcaagggcaa gtggaaacac
     7801 agaaaaaagc ccgcacctga cagtgcgggc tttttttttc gaccaaagga ttgacatgtg
     7861 tcgtggtttg tctggtcaac caccgcggtc tcagtggtgt acggtacaaa ccccgacgtg
     7921 cacag
//
