LOCUS       sequence-179068-        8988 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 8988)
  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 8988)
  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: Bxbi+PhiC31 GFP Bandpass BAC Reporter
FEATURES             Location/Qualifiers
     source          1..8988
                     /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             86..94
                     /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)"
     primer_bind     complement(130..158)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     753..773
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      876..962
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    982..1030
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     protein_bind    2154..2187
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     CDS             complement(2212..2871)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2786..2805
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2872..2974)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      3832..4446
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      3923..4454
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      4522..4741
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             4832..5587
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    5590..5840
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             6166..7341
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             7341..8312
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    8385..8858
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     protein_bind    8923..8980
                     /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 taatagttta cgaggtaagg aggtttaaga tgagtaaagg agaagaactt
      121 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
      181 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
      241 tgcactactg gaaaactacc tgttccatgg ccaacacttg tcactacttt cggttatggt
      301 gttcaatgct ttgcgagata cccagatcat atgaaacagc atgacttttt caagagtgcc
      361 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
      421 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
      481 attgatttta aagaagatgg aaacattctt ggacacaaat tggaatacaa ctataactca
      541 cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagttaa cttcaaaatt
      601 agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca
      661 attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt
      721 tcgaaagatc ccaacgaaaa gagagaccac atggtccttc ttgagtttgt aacagctgct
      781 gggattacac atggcatgga tgatctctac aaataaaagc ttgatatcga attcctgcag
      841 cccgggggat cccatggtac gcgtgctaga ggcatcaaat aaaacgaaag gctcagtcga
      901 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
      961 atccgccgcc ctagaatcga tgcccggatg atcctgacga cggagaccgc cgtcgtcgac
     1021 aagccggccg ttaattaacc tcccgtccgc acagggctcg ctggtgaccg gtccatgttg
     1081 tcggcatacg aggtattgaa gattgggcta agtgccccaa ctggggtaac ctttgagttc
     1141 tctcagttgg ggggctagca aagttaaaca aaattatttg tagagggaaa ccgttgtggt
     1201 ctccctgaat atattatacg agccttatgc atgcccgtaa agttatccag caaccactca
     1261 tagacctagg gcagcagata gggacgacgt ggtgttagct gtgctcgagc tgcaggcgaa
     1321 cacagtggta cgattcatga ttctgcattc gatccaggtt aagagatgca tctcgtatat
     1381 atagtacttg tgcctgtgta acctgagcct tcagcatcgt tcctgcaggg acttgcgaac
     1441 cccctgccaa agtcgtaata taggatcgga caatcctgtc ttcgtggtaa gggttgttgc
     1501 acgttgggga tagcgctagc taacaaagac gcctgctaca acaggagtcg gaaacccgta
     1561 caaagggaac atccacacta cggtgaatcg aagcgcggca tcaggatttc cttttggata
     1621 cctggggcaa agcccatcgt ggtcgaggtg acttggcaca cttacacctg cagcgcgcgc
     1681 atgtggaatt agaggccaag ttcgatccct acaccgacgt acgatgcaac tgtgtggatg
     1741 tgacgagctt catttatacg cttcgcgcgc cggaccggcc tccgcaaggc gcagcagtgc
     1801 acaagcagtc atcaattaac caccgtgtat tcggtatgca gcatcaggca gtttgggcca
     1861 aaaccccagg ggccggttta cacagagacc cgcatcttga cctaactgac aaactgccat
     1921 ggacgactag cccagccaac ggctcttaga cagcccgata cagtgattac cccaggtttg
     1981 cggggcatgg ctacgacttg ctcagctacg tgcgagggca gaaacttttc cgcatttgta
     2041 tgttcaccta tttcagccaa aaaacttaag accgccggtc ttgtccacta ccttgcagta
     2101 atgcggtgga caggatcggc ggttttcttt tctcttctca aatcgatgga gtacgcgccc
     2161 ggggagccca agggcacgcc ctggcacccg cataatggtt tcttagacgt cttacgcccc
     2221 gccctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc
     2281 atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt
     2341 ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa
     2401 atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa
     2461 ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg
     2521 tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg
     2581 ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt
     2641 cattgccata cgaaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc
     2701 cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg
     2761 aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg
     2821 atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc
     2881 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
     2941 atggtgaaag ttggaacctc ttacgtgccg atcaacgtct cattttcgcc aaaagttggc
     3001 ccagggcttc ccggtatcaa cagggacacc aggatttatt tattctgcga agtgatcttc
     3061 cgtcacaggt atttattcgc gataagctca tggagcggcg taaccgtcgc acaggaagga
     3121 cagagaaagc gcggatctgg gaagtgacgg acagaacggt caggacctgg attggggagg
     3181 cggttgccgc cgctgctgct gacggtgtga cgttctctgt tccggtcaca ccacatacgt
     3241 tccgccattc ctatgcgatg cacatgctgt atgccggtat accgctgaaa gttctgcaaa
     3301 gcctgatggg acataagtcc atcagttcaa cggaagtcta cacgaaggtt tttgcgctgg
     3361 atgtggctgc ccggcaccgg gtgcagtttg cgatgccgga gtctgatgcg gttgcgatgc
     3421 tgaaacaatt atcctgagaa taaatgcctt ggcctttata tggaaatgtg gaactgagtg
     3481 gatatgctgt ttttgtctgt taaacagaga agctggctgt tatccactga gaagcgaacg
     3541 aaacagtcgg gaaaatctcc cattatcgta gagatccgca ttattaatct caggagcctg
     3601 tgtagcgttt ataggaagta gtgttctgtc atgatgcctg caagcggtaa cgaaaacgat
     3661 ttgaatatgc cttcaggaac aatagaaatc ttcgtgcggt gttacgttga agtggagcgg
     3721 attatgtcag caatggacag aacaacctaa tgaacacaga accatgatgt ggtctgtcct
     3781 tttacagcca gtagtgctcg ccgcagtcga gcgacagggc gaagccctcg gctggttgcc
     3841 ctcgccgctg ggctggcggc cgtctatggc cctgcaaacg cgccagaaac gccgtcgaag
     3901 ccgtgtgcga gacaccgcgg ccggccgccg gcgttgtgga tacctcgcgg aaaacttggc
     3961 cctcactgac agatgagggg cggacgttga cacttgaggg gccgactcac ccggcgcggc
     4021 gttgacagat gaggggcagg ctcgatttcg gccggcgacg tggagctggc cagcctcgca
     4081 aatcggcgaa aacgcctgat tttacgcgag tttcccacag atgatgtgga caagcctggg
     4141 gataagtgcc ctgcggtatt gacacttgag gggcgcgact actgacagat gaggggcgcg
     4201 atccttgaca cttgaggggc agagtgctga cagatgaggg gcgcacctat tgacatttga
     4261 ggggctgtcc acaggcagaa aatccagcat ttgcaagggt ttccgcccgt ttttcggcca
     4321 ccgctaacct gtcttttaac ctgcttttaa accaatattt ataaaccttg tttttaacca
     4381 gggctgcgcc ctgtgcgcgt gaccgcgcac gccgaagggg ggtgcccccc cttctcgaac
     4441 cctcccggtc gagtgagcga ggaagcacca gggaacagca cttatatatt ctgcttacac
     4501 acgatgcctg aaaaaacttc ccttggggtt atccacttat ccacggggat atttttataa
     4561 ttattttttt tatagttttt agatcttctt ttttagagcg ccttgtaggc ctttatccat
     4621 gctggttcta gagaaggtgt tgtgacaaat tgccctttca gtgtgacaaa tcaccctcaa
     4681 atgacagtcc tgtctgtgac aaattgccct taaccctgtg acaaattgcc ctcagaagaa
     4741 gctgtttttt cacaaagtta tccctgctta ttgactcttt tttatttagt gtgacaatct
     4801 aaaaacttgt cacacttcac atggatctgt catggcggaa acagcggtta tcaatcacaa
     4861 gaaacgtaaa aatagcccgc gaatcgtcca gtcaaacgac ctcactgagg cggcatatag
     4921 tctctcccgg gatcaaaaac gtatgctgta tctgttcgtt gaccagatca gaaaatctga
     4981 tggcacccta caggaacatg acggtatctg cgagatccat gttgctaaat atgctgaaat
     5041 attcggattg acctctgcgg aagccagtaa ggatatacgg caggcattga agagtttcgc
     5101 ggggaaggaa gtggtttttt atcgccctga agaggatgcc ggcgatgaaa aaggctatga
     5161 atcttttcct tggtttatca aacgtgcgca cagtccatcc agagggcttt acagtgtaca
     5221 tatcaaccca tatctcattc ccttctttat cgggttacag aaccggttta cgcagtttcg
     5281 gcttagtgaa acaaaagaaa tcaccaatcc gtatgccatg cgtttatacg aatccctgtg
     5341 tcagtatcgt aagccggatg gctcaggcat cgtctctctg aaaatcgact ggatcataga
     5401 gcgttaccag ctgcctcaaa gttaccagcg tatgcctgac ttccgccgcc gcttcctgca
     5461 ggtctgtgtt aatgagatca acagcagaac tccaatgcgc ctctcataca ttgagaaaaa
     5521 gaaaggccgc cagacgactc atatcgtatt ttccttccgc gatatcactt ccatgacgac
     5581 aggatagtct gagggttatc tgtcacagat ttgagggtgg ttcgtcacat ttgttctgac
     5641 ctactgaggg taatttgtca cagttttgct gtttccttca gcctgcatgg attttctcat
     5701 actttttgaa ctgtaatttt taaggaagcc aaatttgagg gcagtttgtc acagttgatt
     5761 tccttctctt tcccttcgtc atgtgacttg atatcggggg ttagttcgtc atcattgatg
     5821 agggttgatt atcacagttt attactctga attggctatc cgcgtgtgta cctctacctg
     5881 gagtttttcc cacggtggat atttcttctt gcgctgagcg taagagctat ctgacagaac
     5941 agttcttctt tgcttcctcg ccagttcgct cgctatgctc ggttacacgg ctgcggcgag
     6001 cgctagtgat aataagtgac tgaggtatgt gctcttctta tctccttttg tagtgttgct
     6061 cttattttaa acaactttgc ggttttttga tgactttgcg attttgttgt tgctttgcag
     6121 taaattgcaa gatttaataa aaaaacgcaa agcaatgatt aaaggatgtt cagaatgaaa
     6181 ctcatggaaa cacttaacca gtgcataaac gctggtcatg aaatgacgaa ggctatcgcc
     6241 attgcacagt ttaatgatga cagcccggaa gcgaggaaaa taacccggcg ctggagaata
     6301 ggtgaagcag cggatttagt tggggtttct tctcaggcta tcagagatgc cgagaaagca
     6361 gggcgactac cgcacccgga tatggaaatt cgaggacggg ttgagcaacg tgttggttat
     6421 acaattgaac aaattaatca tatgcgtgat gtgtttggta cgcgattgcg acgtgctgaa
     6481 gacgtatttc caccggtgat cggggttgct gcccataaag gtggcgttta caaaacctca
     6541 gtttctgttc atcttgctca ggatctggct ctgaaggggc tacgtgtttt gctcgtggaa
     6601 ggtaacgacc cccagggaac agcctcaatg tatcacggat gggtaccaga tcttcatatt
     6661 catgcagaag acactctcct gcctttctat cttggggaaa aggacgatgt cacttatgca
     6721 ataaagccca cttgctggcc ggggcttgac attattcctt cctgtctggc tctgcaccgt
     6781 attgaaactg agttaatggg caaatttgat gaaggtaaac tgcccaccga tccacacctg
     6841 atgctccgac tggccattga aactgttgct catgactatg atgtcatagt tattgacagc
     6901 gcgcctaacc tgggtatcgg cacgattaat gtcgtatgtg ctgctgatgt gctgattgtt
     6961 cccacgcctg ctgagttgtt tgactacacc tccgcactgc agtttttcga tatgcttcgt
     7021 gatctgctca agaacgttga tcttaaaggg ttcgagcctg atgtacgtat tttgcttacc
     7081 aaatacagca atagtaatgg ctctcagtcc ccgtggatgg aggagcaaat tcgggatgcc
     7141 tggggaagca tggttctaaa aaatgttgta cgtgaaacgg atgaagttgg taaaggtcag
     7201 atccggatga gaactgtttt tgaacaggcc attgatcaac gctcttcaac tggtgcctgg
     7261 agaaatgctc tttctatttg ggaacctgtc tgcaatgaaa ttttcgatcg tctgattaaa
     7321 ccacgctggg agattagata atgaagcgtg cgcctgttat tccaaaacat acgctcaata
     7381 ctcaaccggt tgaagatact tcgttatcga caccagctgc cccgatggtg gattcgttaa
     7441 ttgcgcgcgt aggagtaatg gctcgcggta atgccattac tttgcctgta tgtggtcggg
     7501 atgtgaagtt tactcttgaa gtgctccggg gtgatagtgt tgagaagacc tctcgggtat
     7561 ggtcaggtaa tgaacgtgac caggagctgc ttactgagga cgcactggat gatctcatcc
     7621 cttcttttct actgactggt caacagacac cggcgttcgg tcgaagagta tctggtgtca
     7681 tagaaattgc cgatgggagt cgccgtcgta aagctgctgc acttaccgaa agtgattatc
     7741 gtgttctggt tggcgagctg gatgatgagc agatggctgc attatccaga ttgggtaacg
     7801 attatcgccc aacaagtgct tatgaacgtg gtcagcgtta tgcaagccga ttgcagaatg
     7861 aatttgctgg aaatatttct gcgctggctg atgcggaaaa tatttcacgt aagattatta
     7921 cccgctgtat caacaccgcc aaattgccta aatcagttgt tgctcttttt tctcaccccg
     7981 gtgaactatc tgcccggtca ggtgatgcac ttcaaaaagc ctttacagat aaagaggaat
     8041 tacttaagca gcaggcatct aaccttcatg agcagaaaaa agctggggtg atatttgaag
     8101 ctgaagaagt tatcactctt ttaacttctg tgcttaaaac gtcatctgca tcaagaacta
     8161 gtttaagctc acgacatcag tttgctcctg gagcgacagt attgtataag ggcgataaaa
     8221 tggtgcttaa cctggacagg tcccgtgttc caactgagtg tatagagaaa attgaggcca
     8281 ttcttaagga acttgaaaag ccagcaccct gatgcgacca cgttttagtc tacgtttatc
     8341 tgtctttact taatgtcctt tgttacaggc cagaaagcat aactggcctg aatattctct
     8401 ctgggcccac tgttccactt gtatcgtcgg tctgataatc agactgggac cacggtccca
     8461 ctcgtatcgt cggtctgatt attagtctgg gaccacggtc ccactcgtat cgtcggtctg
     8521 attattagtc tgggaccacg gtcccactcg tatcgtcggt ctgataatca gactgggacc
     8581 acggtcccac tcgtatcgtc ggtctgatta ttagtctggg accatggtcc cactcgtatc
     8641 gtcggtctga ttattagtct gggaccacgg tcccactcgt atcgtcggtc tgattattag
     8701 tctggaacca cggtcccact cgtatcgtcg gtctgattat tagtctggga ccacggtccc
     8761 actcgtatcg tcggtctgat tattagtctg ggaccacgat cccactcgtg ttgtcggtct
     8821 gattatcggt ctgggaccac ggtcccactt gtattgtcga tcagactatc agcgtgagac
     8881 tacgattcca tcaatgcctg tcaagggcaa gtattgacat gtgtcgtggt ttgtctggtc
     8941 aaccaccgcg gtctcagtgg tgtacggtac aaaccccgac gtgcacag
//
