LOCUS       sequence-178112-        7911 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 7911)
  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 7911)
  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 GFP BAC Reporter
FEATURES             Location/Qualifiers
     source          1..7911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(31..59)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     654..674
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      777..863
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    883..931
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     primer_bind     complement(1104..1122)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(1165..1824)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1739..1758
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1825..1927)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      2785..3399
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      2876..3407
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      3475..3694
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             3785..4540
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    4543..4793
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             5119..6294
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             6294..7265
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     protein_bind    7747..7804
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attP site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     misc_RNA        7813..7862
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             7898..7906
                     /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)"
ORIGIN
        1 atgagtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
       61 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga
      121 aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt
      181 gtcactactt tcggttatgg tgttcaatgc tttgcgagat acccagatca tatgaaacag
      241 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc
      301 aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt
      361 aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa
      421 ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca aaagaatgga
      481 atcaaagtta acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac
      541 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
      601 ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt
      661 cttgagtttg taacagctgc tgggattaca catggcatgg atgatctcta caaataaaag
      721 cttgatatcg aattcctgca gcccggggga tcccatggta cgcgtgctag aggcatcaaa
      781 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
      841 acgctctcct gagtaggaca aatccgccgc cctagaatcg atgcccggat gatcctgacg
      901 acggagaccg ccgtcgtcga caagccggcc gttaattaaa aagttaaaca aaattatttg
      961 tagagggaaa ccgttgtggt ctccctgaat atattatacg agccttatgc atgcccgtaa
     1021 agttatccag caaccactca tagacctagg gcagcagata gggacgacgt ggtgttagct
     1081 gtgctcgagg tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     1141 gataataatg gtttcttaga cgtcttacgc cccgccctgc cactcatcgc agtactgttg
     1201 taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat
     1261 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg
     1321 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg
     1381 attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc
     1441 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta
     1501 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg
     1561 aacactatcc catatcacca gctcaccgtc tttcattgcc atacgaaatt ccggatgagc
     1621 attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt
     1681 tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc
     1741 aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt
     1801 atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc
     1861 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     1921 ccgatcaacg tctcattttc gccaaaagtt ggcccagggc ttcccggtat caacagggac
     1981 accaggattt atttattctg cgaagtgatc ttccgtcaca ggtatttatt cgcgataagc
     2041 tcatggagcg gcgtaaccgt cgcacaggaa ggacagagaa agcgcggatc tgggaagtga
     2101 cggacagaac ggtcaggacc tggattgggg aggcggttgc cgccgctgct gctgacggtg
     2161 tgacgttctc tgttccggtc acaccacata cgttccgcca ttcctatgcg atgcacatgc
     2221 tgtatgccgg tataccgctg aaagttctgc aaagcctgat gggacataag tccatcagtt
     2281 caacggaagt ctacacgaag gtttttgcgc tggatgtggc tgcccggcac cgggtgcagt
     2341 ttgcgatgcc ggagtctgat gcggttgcga tgctgaaaca attatcctga gaataaatgc
     2401 cttggccttt atatggaaat gtggaactga gtggatatgc tgtttttgtc tgttaaacag
     2461 agaagctggc tgttatccac tgagaagcga acgaaacagt cgggaaaatc tcccattatc
     2521 gtagagatcc gcattattaa tctcaggagc ctgtgtagcg tttataggaa gtagtgttct
     2581 gtcatgatgc ctgcaagcgg taacgaaaac gatttgaata tgccttcagg aacaatagaa
     2641 atcttcgtgc ggtgttacgt tgaagtggag cggattatgt cagcaatgga cagaacaacc
     2701 taatgaacac agaaccatga tgtggtctgt ccttttacag ccagtagtgc tcgccgcagt
     2761 cgagcgacag ggcgaagccc tcggctggtt gccctcgccg ctgggctggc ggccgtctat
     2821 ggccctgcaa acgcgccaga aacgccgtcg aagccgtgtg cgagacaccg cggccggccg
     2881 ccggcgttgt ggatacctcg cggaaaactt ggccctcact gacagatgag gggcggacgt
     2941 tgacacttga ggggccgact cacccggcgc ggcgttgaca gatgaggggc aggctcgatt
     3001 tcggccggcg acgtggagct ggccagcctc gcaaatcggc gaaaacgcct gattttacgc
     3061 gagtttccca cagatgatgt ggacaagcct ggggataagt gccctgcggt attgacactt
     3121 gaggggcgcg actactgaca gatgaggggc gcgatccttg acacttgagg ggcagagtgc
     3181 tgacagatga ggggcgcacc tattgacatt tgaggggctg tccacaggca gaaaatccag
     3241 catttgcaag ggtttccgcc cgtttttcgg ccaccgctaa cctgtctttt aacctgcttt
     3301 taaaccaata tttataaacc ttgtttttaa ccagggctgc gccctgtgcg cgtgaccgcg
     3361 cacgccgaag gggggtgccc ccccttctcg aaccctcccg gtcgagtgag cgaggaagca
     3421 ccagggaaca gcacttatat attctgctta cacacgatgc ctgaaaaaac ttcccttggg
     3481 gttatccact tatccacggg gatattttta taattatttt ttttatagtt tttagatctt
     3541 cttttttaga gcgccttgta ggcctttatc catgctggtt ctagagaagg tgttgtgaca
     3601 aattgccctt tcagtgtgac aaatcaccct caaatgacag tcctgtctgt gacaaattgc
     3661 ccttaaccct gtgacaaatt gccctcagaa gaagctgttt tttcacaaag ttatccctgc
     3721 ttattgactc ttttttattt agtgtgacaa tctaaaaact tgtcacactt cacatggatc
     3781 tgtcatggcg gaaacagcgg ttatcaatca caagaaacgt aaaaatagcc cgcgaatcgt
     3841 ccagtcaaac gacctcactg aggcggcata tagtctctcc cgggatcaaa aacgtatgct
     3901 gtatctgttc gttgaccaga tcagaaaatc tgatggcacc ctacaggaac atgacggtat
     3961 ctgcgagatc catgttgcta aatatgctga aatattcgga ttgacctctg cggaagccag
     4021 taaggatata cggcaggcat tgaagagttt cgcggggaag gaagtggttt tttatcgccc
     4081 tgaagaggat gccggcgatg aaaaaggcta tgaatctttt ccttggttta tcaaacgtgc
     4141 gcacagtcca tccagagggc tttacagtgt acatatcaac ccatatctca ttcccttctt
     4201 tatcgggtta cagaaccggt ttacgcagtt tcggcttagt gaaacaaaag aaatcaccaa
     4261 tccgtatgcc atgcgtttat acgaatccct gtgtcagtat cgtaagccgg atggctcagg
     4321 catcgtctct ctgaaaatcg actggatcat agagcgttac cagctgcctc aaagttacca
     4381 gcgtatgcct gacttccgcc gccgcttcct gcaggtctgt gttaatgaga tcaacagcag
     4441 aactccaatg cgcctctcat acattgagaa aaagaaaggc cgccagacga ctcatatcgt
     4501 attttccttc cgcgatatca cttccatgac gacaggatag tctgagggtt atctgtcaca
     4561 gatttgaggg tggttcgtca catttgttct gacctactga gggtaatttg tcacagtttt
     4621 gctgtttcct tcagcctgca tggattttct catacttttt gaactgtaat ttttaaggaa
     4681 gccaaatttg agggcagttt gtcacagttg atttccttct ctttcccttc gtcatgtgac
     4741 ttgatatcgg gggttagttc gtcatcattg atgagggttg attatcacag tttattactc
     4801 tgaattggct atccgcgtgt gtacctctac ctggagtttt tcccacggtg gatatttctt
     4861 cttgcgctga gcgtaagagc tatctgacag aacagttctt ctttgcttcc tcgccagttc
     4921 gctcgctatg ctcggttaca cggctgcggc gagcgctagt gataataagt gactgaggta
     4981 tgtgctcttc ttatctcctt ttgtagtgtt gctcttattt taaacaactt tgcggttttt
     5041 tgatgacttt gcgattttgt tgttgctttg cagtaaattg caagatttaa taaaaaaacg
     5101 caaagcaatg attaaaggat gttcagaatg aaactcatgg aaacacttaa ccagtgcata
     5161 aacgctggtc atgaaatgac gaaggctatc gccattgcac agtttaatga tgacagcccg
     5221 gaagcgagga aaataacccg gcgctggaga ataggtgaag cagcggattt agttggggtt
     5281 tcttctcagg ctatcagaga tgccgagaaa gcagggcgac taccgcaccc ggatatggaa
     5341 attcgaggac gggttgagca acgtgttggt tatacaattg aacaaattaa tcatatgcgt
     5401 gatgtgtttg gtacgcgatt gcgacgtgct gaagacgtat ttccaccggt gatcggggtt
     5461 gctgcccata aaggtggcgt ttacaaaacc tcagtttctg ttcatcttgc tcaggatctg
     5521 gctctgaagg ggctacgtgt tttgctcgtg gaaggtaacg acccccaggg aacagcctca
     5581 atgtatcacg gatgggtacc agatcttcat attcatgcag aagacactct cctgcctttc
     5641 tatcttgggg aaaaggacga tgtcacttat gcaataaagc ccacttgctg gccggggctt
     5701 gacattattc cttcctgtct ggctctgcac cgtattgaaa ctgagttaat gggcaaattt
     5761 gatgaaggta aactgcccac cgatccacac ctgatgctcc gactggccat tgaaactgtt
     5821 gctcatgact atgatgtcat agttattgac agcgcgccta acctgggtat cggcacgatt
     5881 aatgtcgtat gtgctgctga tgtgctgatt gttcccacgc ctgctgagtt gtttgactac
     5941 acctccgcac tgcagttttt cgatatgctt cgtgatctgc tcaagaacgt tgatcttaaa
     6001 gggttcgagc ctgatgtacg tattttgctt accaaataca gcaatagtaa tggctctcag
     6061 tccccgtgga tggaggagca aattcgggat gcctggggaa gcatggttct aaaaaatgtt
     6121 gtacgtgaaa cggatgaagt tggtaaaggt cagatccgga tgagaactgt ttttgaacag
     6181 gccattgatc aacgctcttc aactggtgcc tggagaaatg ctctttctat ttgggaacct
     6241 gtctgcaatg aaattttcga tcgtctgatt aaaccacgct gggagattag ataatgaagc
     6301 gtgcgcctgt tattccaaaa catacgctca atactcaacc ggttgaagat acttcgttat
     6361 cgacaccagc tgccccgatg gtggattcgt taattgcgcg cgtaggagta atggctcgcg
     6421 gtaatgccat tactttgcct gtatgtggtc gggatgtgaa gtttactctt gaagtgctcc
     6481 ggggtgatag tgttgagaag acctctcggg tatggtcagg taatgaacgt gaccaggagc
     6541 tgcttactga ggacgcactg gatgatctca tcccttcttt tctactgact ggtcaacaga
     6601 caccggcgtt cggtcgaaga gtatctggtg tcatagaaat tgccgatggg agtcgccgtc
     6661 gtaaagctgc tgcacttacc gaaagtgatt atcgtgttct ggttggcgag ctggatgatg
     6721 agcagatggc tgcattatcc agattgggta acgattatcg cccaacaagt gcttatgaac
     6781 gtggtcagcg ttatgcaagc cgattgcaga atgaatttgc tggaaatatt tctgcgctgg
     6841 ctgatgcgga aaatatttca cgtaagatta ttacccgctg tatcaacacc gccaaattgc
     6901 ctaaatcagt tgttgctctt ttttctcacc ccggtgaact atctgcccgg tcaggtgatg
     6961 cacttcaaaa agcctttaca gataaagagg aattacttaa gcagcaggca tctaaccttc
     7021 atgagcagaa aaaagctggg gtgatatttg aagctgaaga agttatcact cttttaactt
     7081 ctgtgcttaa aacgtcatct gcatcaagaa ctagtttaag ctcacgacat cagtttgctc
     7141 ctggagcgac agtattgtat aagggcgata aaatggtgct taacctggac aggtcccgtg
     7201 ttccaactga gtgtatagag aaaattgagg ccattcttaa ggaacttgaa aagccagcac
     7261 cctgatgcga ccacgtttta gtctacgttt atctgtcttt acttaatgtc ctttgttaca
     7321 ggccagaaag cataactggc ctgaatattc tctctgggcc cactgttcca cttgtatcgt
     7381 cggtctgata atcagactgg gaccacggtc ccactcgtat cgtcggtctg attattagtc
     7441 tgggaccacg gtcccactcg tatcgtcggt ctgattatta gtctgggacc acggtcccac
     7501 tcgtatcgtc ggtctgatta ttagtctgga accacggtcc cactcgtatc gtcggtctga
     7561 ttattagtct gggaccacgg tcccactcgt atcgtcggtc tgattattag tctgggacca
     7621 cgatcccact cgtgttgtcg gtctgattat cggtctggga ccacggtccc acttgtattg
     7681 tcgatcagac tatcagcgtg agactacgat tccatcaatg cctgtcaagg gcaagtattg
     7741 acatgtgtcg tggtttgtct ggtcaaccac cgcggtctca gtggtgtacg gtacaaaccc
     7801 cgacgtgcac agctgtcacc ggatgtgctt tccggtctga tgagtccgtg aggacgaaac
     7861 agcctctaca aataattttg tttaatagtt tacgaggtaa ggaggtttaa g
//
