LOCUS       sequence-178114-        8956 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  synthetic gene circuit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8956)
  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 8956)
  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: Mixed-signal integration BAC Reporter
FEATURES             Location/Qualifiers
     source          1..8956
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(254..1225)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(1225..2400)
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     misc_feature    2726..2976
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(2979..3734)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      3825..4044
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      4112..4643
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      4120..4734
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     promoter        5592..5694
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5695..6354
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5761..5780)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    7549..7582
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    7597..7645
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     terminator      complement(7665..7751)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(7854..7874)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     8469..8497
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     RBS             8533..8541
                     /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)"
     misc_RNA        complement(8577..8626)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     protein_bind    8635..8692
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attP site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
ORIGIN
        1 atcagaccga cgatacgagt gggaccgtgg tcccagacta ataatcagac cgacgatacg
       61 agtgggaccg tggtcccaga ctaataatca gaccgacgat acgagtggga ccgtggtccc
      121 agtctgatta tcagaccgac gatacaagtg gaacagtggg cccagagaga atattcaggc
      181 cagttatgct ttctggcctg taacaaagga cattaagtaa agacagataa acgtagacta
      241 aaacgtggtc gcatcagggt gctggctttt caagttcctt aagaatggcc tcaattttct
      301 ctatacactc agttggaaca cgggacctgt ccaggttaag caccatttta tcgcccttat
      361 acaatactgt cgctccagga gcaaactgat gtcgtgagct taaactagtt cttgatgcag
      421 atgacgtttt aagcacagaa gttaaaagag tgataacttc ttcagcttca aatatcaccc
      481 cagctttttt ctgctcatga aggttagatg cctgctgctt aagtaattcc tctttatctg
      541 taaaggcttt ttgaagtgca tcacctgacc gggcagatag ttcaccgggg tgagaaaaaa
      601 gagcaacaac tgatttaggc aatttggcgg tgttgataca gcgggtaata atcttacgtg
      661 aaatattttc cgcatcagcc agcgcagaaa tatttccagc aaattcattc tgcaatcggc
      721 ttgcataacg ctgaccacgt tcataagcac ttgttgggcg ataatcgtta cccaatctgg
      781 ataatgcagc catctgctca tcatccagct cgccaaccag aacacgataa tcactttcgg
      841 taagtgcagc agctttacga cggcgactcc catcggcaat ttctatgaca ccagatactc
      901 ttcgaccgaa cgccggtgtc tgttgaccag tcagtagaaa agaagggatg agatcatcca
      961 gtgcgtcctc agtaagcagc tcctggtcac gttcattacc tgaccatacc cgagaggtct
     1021 tctcaacact atcaccccgg agcacttcaa gagtaaactt cacatcccga ccacatacag
     1081 gcaaagtaat ggcattaccg cgagccatta ctcctacgcg cgcaattaac gaatccacca
     1141 tcggggcagc tggtgtcgat aacgaagtat cttcaaccgg ttgagtattg agcgtatgtt
     1201 ttggaataac aggcgcacgc ttcattatct aatctcccag cgtggtttaa tcagacgatc
     1261 gaaaatttca ttgcagacag gttcccaaat agaaagagca tttctccagg caccagttga
     1321 agagcgttga tcaatggcct gttcaaaaac agttctcatc cggatctgac ctttaccaac
     1381 ttcatccgtt tcacgtacaa cattttttag aaccatgctt ccccaggcat cccgaatttg
     1441 ctcctccatc cacggggact gagagccatt actattgctg tatttggtaa gcaaaatacg
     1501 tacatcaggc tcgaaccctt taagatcaac gttcttgagc agatcacgaa gcatatcgaa
     1561 aaactgcagt gcggaggtgt agtcaaacaa ctcagcaggc gtgggaacaa tcagcacatc
     1621 agcagcacat acgacattaa tcgtgccgat acccaggtta ggcgcgctgt caataactat
     1681 gacatcatag tcatgagcaa cagtttcaat ggccagtcgg agcatcaggt gtggatcggt
     1741 gggcagttta ccttcatcaa atttgcccat taactcagtt tcaatacggt gcagagccag
     1801 acaggaagga ataatgtcaa gccccggcca gcaagtgggc tttattgcat aagtgacatc
     1861 gtccttttcc ccaagataga aaggcaggag agtgtcttct gcatgaatat gaagatctgg
     1921 tacccatccg tgatacattg aggctgttcc ctgggggtcg ttaccttcca cgagcaaaac
     1981 acgtagcccc ttcagagcca gatcctgagc aagatgaaca gaaactgagg ttttgtaaac
     2041 gccaccttta tgggcagcaa ccccgatcac cggtggaaat acgtcttcag cacgtcgcaa
     2101 tcgcgtacca aacacatcac gcatatgatt aatttgttca attgtataac caacacgttg
     2161 ctcaacccgt cctcgaattt ccatatccgg gtgcggtagt cgccctgctt tctcggcatc
     2221 tctgatagcc tgagaagaaa ccccaactaa atccgctgct tcacctattc tccagcgccg
     2281 ggttattttc ctcgcttccg ggctgtcatc attaaactgt gcaatggcga tagccttcgt
     2341 catttcatga ccagcgttta tgcactggtt aagtgtttcc atgagtttca ttctgaacat
     2401 cctttaatca ttgctttgcg tttttttatt aaatcttgca atttactgca aagcaacaac
     2461 aaaatcgcaa agtcatcaaa aaaccgcaaa gttgtttaaa ataagagcaa cactacaaaa
     2521 ggagataaga agagcacata cctcagtcac ttattatcac tagcgctcgc cgcagccgtg
     2581 taaccgagca tagcgagcga actggcgagg aagcaaagaa gaactgttct gtcagatagc
     2641 tcttacgctc agcgcaagaa gaaatatcca ccgtgggaaa aactccaggt agaggtacac
     2701 acgcggatag ccaattcaga gtaataaact gtgataatca accctcatca atgatgacga
     2761 actaaccccc gatatcaagt cacatgacga agggaaagag aaggaaatca actgtgacaa
     2821 actgccctca aatttggctt ccttaaaaat tacagttcaa aaagtatgag aaaatccatg
     2881 caggctgaag gaaacagcaa aactgtgaca aattaccctc agtaggtcag aacaaatgtg
     2941 acgaaccacc ctcaaatctg tgacagataa ccctcagact atcctgtcgt catggaagtg
     3001 atatcgcgga aggaaaatac gatatgagtc gtctggcggc ctttcttttt ctcaatgtat
     3061 gagaggcgca ttggagttct gctgttgatc tcattaacac agacctgcag gaagcggcgg
     3121 cggaagtcag gcatacgctg gtaactttga ggcagctggt aacgctctat gatccagtcg
     3181 attttcagag agacgatgcc tgagccatcc ggcttacgat actgacacag ggattcgtat
     3241 aaacgcatgg catacggatt ggtgatttct tttgtttcac taagccgaaa ctgcgtaaac
     3301 cggttctgta acccgataaa gaagggaatg agatatgggt tgatatgtac actgtaaagc
     3361 cctctggatg gactgtgcgc acgtttgata aaccaaggaa aagattcata gcctttttca
     3421 tcgccggcat cctcttcagg gcgataaaaa accacttcct tccccgcgaa actcttcaat
     3481 gcctgccgta tatccttact ggcttccgca gaggtcaatc cgaatatttc agcatattta
     3541 gcaacatgga tctcgcagat accgtcatgt tcctgtaggg tgccatcaga ttttctgatc
     3601 tggtcaacga acagatacag catacgtttt tgatcccggg agagactata tgccgcctca
     3661 gtgaggtcgt ttgactggac gattcgcggg ctatttttac gtttcttgtg attgataacc
     3721 gctgtttccg ccatgacaga tccatgtgaa gtgtgacaag tttttagatt gtcacactaa
     3781 ataaaaaaga gtcaataagc agggataact ttgtgaaaaa acagcttctt ctgagggcaa
     3841 tttgtcacag ggttaagggc aatttgtcac agacaggact gtcatttgag ggtgatttgt
     3901 cacactgaaa gggcaatttg tcacaacacc ttctctagaa ccagcatgga taaaggccta
     3961 caaggcgctc taaaaaagaa gatctaaaaa ctataaaaaa aataattata aaaatatccc
     4021 cgtggataag tggataaccc caagggaagt tttttcaggc atcgtgtgta agcagaatat
     4081 ataagtgctg ttccctggtg cttcctcgct cactcgaccg ggagggttcg agaagggggg
     4141 gcacccccct tcggcgtgcg cggtcacgcg cacagggcgc agccctggtt aaaaacaagg
     4201 tttataaata ttggtttaaa agcaggttaa aagacaggtt agcggtggcc gaaaaacggg
     4261 cggaaaccct tgcaaatgct ggattttctg cctgtggaca gcccctcaaa tgtcaatagg
     4321 tgcgcccctc atctgtcagc actctgcccc tcaagtgtca aggatcgcgc ccctcatctg
     4381 tcagtagtcg cgcccctcaa gtgtcaatac cgcagggcac ttatccccag gcttgtccac
     4441 atcatctgtg ggaaactcgc gtaaaatcag gcgttttcgc cgatttgcga ggctggccag
     4501 ctccacgtcg ccggccgaaa tcgagcctgc ccctcatctg tcaacgccgc gccgggtgag
     4561 tcggcccctc aagtgtcaac gtccgcccct catctgtcag tgagggccaa gttttccgcg
     4621 aggtatccac aacgccggcg gccggccgcg gtgtctcgca cacggcttcg acggcgtttc
     4681 tggcgcgttt gcagggccat agacggccgc cagcccagcg gcgagggcaa ccagccgagg
     4741 gcttcgccct gtcgctcgac tgcggcgagc actactggct gtaaaaggac agaccacatc
     4801 atggttctgt gttcattagg ttgttctgtc cattgctgac ataatccgct ccacttcaac
     4861 gtaacaccgc acgaagattt ctattgttcc tgaaggcata ttcaaatcgt tttcgttacc
     4921 gcttgcaggc atcatgacag aacactactt cctataaacg ctacacaggc tcctgagatt
     4981 aataatgcgg atctctacga taatgggaga ttttcccgac tgtttcgttc gcttctcagt
     5041 ggataacagc cagcttctct gtttaacaga caaaaacagc atatccactc agttccacat
     5101 ttccatataa aggccaaggc atttattctc aggataattg tttcagcatc gcaaccgcat
     5161 cagactccgg catcgcaaac tgcacccggt gccgggcagc cacatccagc gcaaaaacct
     5221 tcgtgtagac ttccgttgaa ctgatggact tatgtcccat caggctttgc agaactttca
     5281 gcggtatacc ggcatacagc atgtgcatcg cataggaatg gcggaacgta tgtggtgtga
     5341 ccggaacaga gaacgtcaca ccgtcagcag cagcggcggc aaccgcctcc ccaatccagg
     5401 tcctgaccgt tctgtccgtc acttcccaga tccgcgcttt ctctgtcctt cctgtgcgac
     5461 ggttacgccg ctccatgagc ttatcgcgaa taaatacctg tgacggaaga tcacttcgca
     5521 gaataaataa atcctggtgt ccctgttgat accgggaagc cctgggccaa cttttggcga
     5581 aaatgagacg ttgatcggca cgtaagaggt tccaactttc accataatga aataagatca
     5641 ctaccgggcg tattttttga gttatcgaga ttttcaggag ctaaggaagc taaaatggag
     5701 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
     5761 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
     5821 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
     5881 cttgcccgcc tgatgaatgc tcatccggaa tttcgtatgg caatgaaaga cggtgagctg
     5941 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
     6001 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
     6061 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
     6121 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
     6181 atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag
     6241 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
     6301 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaagacgtc
     6361 taagaaacca ttaatgccaa cacaattaac atctcaatca aggtaaatgc tttttgcttt
     6421 ttttgcatcg atttgagaag agaaaagaaa accgccgatc ctgtccaccg cattactgca
     6481 aggtagtgga caagaccggc ggtcttaagt tttttggctg aaataggtga acatacaaat
     6541 gcggaaaagt ttctgccctc gcacgtagct gagcaagtcg tagccatgcc ccgcaaacct
     6601 ggggtaatca ctgtatcggg ctgtctaaga gccgttggct gggctagtcg tccatggcag
     6661 tttgtcagtt aggtcaagat gcgggtctct gtgtaaaccg gcccctgggg ttttggccca
     6721 aactgcctga tgctgcatac cgaatacacg gtggttaatt gatgactgct tgtgcactgc
     6781 tgcgccttgc ggaggccggt ccggcgcgcg aagcgtataa atgaagctcg tcacatccac
     6841 acagttgcat cgtacgtcgg tgtagggatc gaacttggcc tctaattcca catgcgcgcg
     6901 ctgcaggtgt aagtgtgcca agtcacctcg accacgatgg gctttgcccc aggtatccaa
     6961 aaggaaatcc tgatgccgcg cttcgattca ccgtagtgtg gatgttccct ttgtacgggt
     7021 ttccgactcc tgttgtagca ggcgtctttg ttagctagcg ctatccccaa cgtgcaacaa
     7081 cccttaccac gaagacagga ttgtccgatc ctatattacg actttggcag ggggttcgca
     7141 agtccctgca ggaacgatgc tgaaggctca ggttacacag gcacaagtac tatatatacg
     7201 agatgcatct cttaacctgg atcgaatgca gaatcatgaa tcgtaccact gtgttcgcct
     7261 gcagctcgag cacagctaac accacgtcgt ccctatctgc tgccctaggt ctatgagtgg
     7321 ttgctggata actttacggg catgcataag gctcgtataa tatattcagg gagaccacaa
     7381 cggtttccct ctacaaataa ttttgtttaa ctttgctagc aaaggagttt tttagttacc
     7441 ttaattgaaa taaacgaaat aaaaactcgc ttagcccaat cttcaatacc tcgtatgccg
     7501 acaacatgga ccggtcacca gcgagccctg tgcggacggg aggtgcgggt gccagggcgt
     7561 gcccttgggc tccccgggcg cgtactcctt aattaacggc cggcttgtcg acgacggcgg
     7621 tctccgtcgt caggatcatc cgggcatcga ttctagggcg gcggatttgt cctactcagg
     7681 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt
     7741 cgttttattt gatgcctcta gcacgcgtac catgggatcc cccgggctgc aggaattcga
     7801 tatcaagctt ttatttgtag agatcatcca tgccatgtgt aatcccagca gctgttacaa
     7861 actcaagaag gaccatgtgg tctctctttt cgttgggatc tttcgaaagg gcagattgtg
     7921 tggacaggta atggttgtct ggtaaaagga cagggccatc gccaattgga gtattttgtt
     7981 gataatggtc tgctagttga acgcttccat cttcaatgtt gtgtctaatt ttgaagttaa
     8041 ctttgattcc attcttttgt ttgtctgcca tgatgtatac attgtgtgag ttatagttgt
     8101 attccaattt gtgtccaaga atgtttccat cttctttaaa atcaatacct tttaactcga
     8161 ttctattaac aagggtatca ccttcaaact tgacttcagc acgtgtcttg tagttcccgt
     8221 catctttgaa aaatatagtt ctttcctgta cataaccttc gggcatggca ctcttgaaaa
     8281 agtcatgctg tttcatatga tctgggtatc tcgcaaagca ttgaacacca taaccgaaag
     8341 tagtgacaag tgttggccat ggaacaggta gttttccagt agtgcaaata aatttaaggg
     8401 taagttttcc gtatgttgca tcaccttcac cctctccact gacagaaaat ttgtgcccat
     8461 taacatcacc atctaattca acaagaattg ggacaactcc agtgaaaagt tcttctcctt
     8521 tactcatctt aaacctcctt acctcgtaaa ctattaaaca aaattatttg tagaggctgt
     8581 ttcgtcctca cggactcatc agaccggaaa gcacatccgg tgacagctgt gcacgtcggg
     8641 gtttgtaccg tacaccactg agaccgcggt ggttgaccag acaaaccacg acacatgtcc
     8701 cccaactgag agaactcaaa ggttacccca gttggggcac cgagcaccgt acttgccctt
     8761 gacaggcatt gatggaatcg tagtctcacg ctgatagtct gatcgacaat acaagtggga
     8821 ccgtggtccc agaccgataa tcagaccgac aacacgagtg ggatcgtggt cccagactaa
     8881 taatcagacc gacgatacga gtgggaccgt ggtcccagac taataatcag accgacgata
     8941 cgagtgggac cgtggt
//
