LOCUS       sequence-178113-        9004 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 9004)
  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 9004)
  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+TP901 GFP Bandpass BAC Reporter
FEATURES             Location/Qualifiers
     source          1..9004
                     /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)"
     CDS             complement(2129..2788)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2703..2722
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2789..2891)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      3749..4363
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      3840..4371
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      4439..4658
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             4749..5504
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    5507..5757
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             6083..7258
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             7258..8229
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    8302..8775
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     protein_bind    8840..8897
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attP site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     misc_RNA        8906..8955
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             8991..8999
                     /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 gttaattaac ctcccgtccg cacagggctc
      961 gctggtgacc ggtccatgtt gtcggcatac gaggtattga agattgggct aagcgagttt
     1021 ttatttcgtt tatttcaatt aaggtaacta aaaaactcct ttgctagcaa agttaaacaa
     1081 aattatttgt agagggaaac cgttgtggtc tccctgaata tattatacga gccttatgca
     1141 tgcccgtaaa gttatccagc aaccactcat agacctaggg cagcagatag ggacgacgtg
     1201 gtgttagctg tgctcgagct gcaggcgaac acagtggtac gattcatgat tctgcattcg
     1261 atccaggtta agagatgcat ctcgtatata tagtacttgt gcctgtgtaa cctgagcctt
     1321 cagcatcgtt cctgcaggga cttgcgaacc ccctgccaaa gtcgtaatat aggatcggac
     1381 aatcctgtct tcgtggtaag ggttgttgca cgttggggat agcgctagct aacaaagacg
     1441 cctgctacaa caggagtcgg aaacccgtac aaagggaaca tccacactac ggtgaatcga
     1501 agcgcggcat caggatttcc ttttggatac ctggggcaaa gcccatcgtg gtcgaggtga
     1561 cttggcacac ttacacctgc agcgcgcgca tgtggaatta gaggccaagt tcgatcccta
     1621 caccgacgta cgatgcaact gtgtggatgt gacgagcttc atttatacgc ttcgcgcgcc
     1681 ggaccggcct ccgcaaggcg cagcagtgca caagcagtca tcaattaacc accgtgtatt
     1741 cggtatgcag catcaggcag tttgggccaa aaccccaggg gccggtttac acagagaccc
     1801 gcatcttgac ctaactgaca aactgccatg gacgactagc ccagccaacg gctcttagac
     1861 agcccgatac agtgattacc ccaggtttgc ggggcatggc tacgacttgc tcagctacgt
     1921 gcgagggcag aaacttttcc gcatttgtat gttcacctat ttcagccaaa aaacttaaga
     1981 ccgccggtct tgtccactac cttgcagtaa tgcggtggac aggatcggcg gttttctttt
     2041 ctcttctcaa atcgatgcaa aaaaagcaaa aagcatttac cttgattgag atgttaattg
     2101 tgttggcatt aatggtttct tagacgtctt acgccccgcc ctgccactca tcgcagtact
     2161 gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct
     2221 gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa
     2281 cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc
     2341 agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt
     2401 tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt
     2461 ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag
     2521 ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacga aattccggat
     2581 gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt
     2641 tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt
     2701 gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg
     2761 tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata
     2821 actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta
     2881 cgtgccgatc aacgtctcat tttcgccaaa agttggccca gggcttcccg gtatcaacag
     2941 ggacaccagg atttatttat tctgcgaagt gatcttccgt cacaggtatt tattcgcgat
     3001 aagctcatgg agcggcgtaa ccgtcgcaca ggaaggacag agaaagcgcg gatctgggaa
     3061 gtgacggaca gaacggtcag gacctggatt ggggaggcgg ttgccgccgc tgctgctgac
     3121 ggtgtgacgt tctctgttcc ggtcacacca catacgttcc gccattccta tgcgatgcac
     3181 atgctgtatg ccggtatacc gctgaaagtt ctgcaaagcc tgatgggaca taagtccatc
     3241 agttcaacgg aagtctacac gaaggttttt gcgctggatg tggctgcccg gcaccgggtg
     3301 cagtttgcga tgccggagtc tgatgcggtt gcgatgctga aacaattatc ctgagaataa
     3361 atgccttggc ctttatatgg aaatgtggaa ctgagtggat atgctgtttt tgtctgttaa
     3421 acagagaagc tggctgttat ccactgagaa gcgaacgaaa cagtcgggaa aatctcccat
     3481 tatcgtagag atccgcatta ttaatctcag gagcctgtgt agcgtttata ggaagtagtg
     3541 ttctgtcatg atgcctgcaa gcggtaacga aaacgatttg aatatgcctt caggaacaat
     3601 agaaatcttc gtgcggtgtt acgttgaagt ggagcggatt atgtcagcaa tggacagaac
     3661 aacctaatga acacagaacc atgatgtggt ctgtcctttt acagccagta gtgctcgccg
     3721 cagtcgagcg acagggcgaa gccctcggct ggttgccctc gccgctgggc tggcggccgt
     3781 ctatggccct gcaaacgcgc cagaaacgcc gtcgaagccg tgtgcgagac accgcggccg
     3841 gccgccggcg ttgtggatac ctcgcggaaa acttggccct cactgacaga tgaggggcgg
     3901 acgttgacac ttgaggggcc gactcacccg gcgcggcgtt gacagatgag gggcaggctc
     3961 gatttcggcc ggcgacgtgg agctggccag cctcgcaaat cggcgaaaac gcctgatttt
     4021 acgcgagttt cccacagatg atgtggacaa gcctggggat aagtgccctg cggtattgac
     4081 acttgagggg cgcgactact gacagatgag gggcgcgatc cttgacactt gaggggcaga
     4141 gtgctgacag atgaggggcg cacctattga catttgaggg gctgtccaca ggcagaaaat
     4201 ccagcatttg caagggtttc cgcccgtttt tcggccaccg ctaacctgtc ttttaacctg
     4261 cttttaaacc aatatttata aaccttgttt ttaaccaggg ctgcgccctg tgcgcgtgac
     4321 cgcgcacgcc gaaggggggt gccccccctt ctcgaaccct cccggtcgag tgagcgagga
     4381 agcaccaggg aacagcactt atatattctg cttacacacg atgcctgaaa aaacttccct
     4441 tggggttatc cacttatcca cggggatatt tttataatta ttttttttat agtttttaga
     4501 tcttcttttt tagagcgcct tgtaggcctt tatccatgct ggttctagag aaggtgttgt
     4561 gacaaattgc cctttcagtg tgacaaatca ccctcaaatg acagtcctgt ctgtgacaaa
     4621 ttgcccttaa ccctgtgaca aattgccctc agaagaagct gttttttcac aaagttatcc
     4681 ctgcttattg actctttttt atttagtgtg acaatctaaa aacttgtcac acttcacatg
     4741 gatctgtcat ggcggaaaca gcggttatca atcacaagaa acgtaaaaat agcccgcgaa
     4801 tcgtccagtc aaacgacctc actgaggcgg catatagtct ctcccgggat caaaaacgta
     4861 tgctgtatct gttcgttgac cagatcagaa aatctgatgg caccctacag gaacatgacg
     4921 gtatctgcga gatccatgtt gctaaatatg ctgaaatatt cggattgacc tctgcggaag
     4981 ccagtaagga tatacggcag gcattgaaga gtttcgcggg gaaggaagtg gttttttatc
     5041 gccctgaaga ggatgccggc gatgaaaaag gctatgaatc ttttccttgg tttatcaaac
     5101 gtgcgcacag tccatccaga gggctttaca gtgtacatat caacccatat ctcattccct
     5161 tctttatcgg gttacagaac cggtttacgc agtttcggct tagtgaaaca aaagaaatca
     5221 ccaatccgta tgccatgcgt ttatacgaat ccctgtgtca gtatcgtaag ccggatggct
     5281 caggcatcgt ctctctgaaa atcgactgga tcatagagcg ttaccagctg cctcaaagtt
     5341 accagcgtat gcctgacttc cgccgccgct tcctgcaggt ctgtgttaat gagatcaaca
     5401 gcagaactcc aatgcgcctc tcatacattg agaaaaagaa aggccgccag acgactcata
     5461 tcgtattttc cttccgcgat atcacttcca tgacgacagg atagtctgag ggttatctgt
     5521 cacagatttg agggtggttc gtcacatttg ttctgaccta ctgagggtaa tttgtcacag
     5581 ttttgctgtt tccttcagcc tgcatggatt ttctcatact ttttgaactg taatttttaa
     5641 ggaagccaaa tttgagggca gtttgtcaca gttgatttcc ttctctttcc cttcgtcatg
     5701 tgacttgata tcgggggtta gttcgtcatc attgatgagg gttgattatc acagtttatt
     5761 actctgaatt ggctatccgc gtgtgtacct ctacctggag tttttcccac ggtggatatt
     5821 tcttcttgcg ctgagcgtaa gagctatctg acagaacagt tcttctttgc ttcctcgcca
     5881 gttcgctcgc tatgctcggt tacacggctg cggcgagcgc tagtgataat aagtgactga
     5941 ggtatgtgct cttcttatct ccttttgtag tgttgctctt attttaaaca actttgcggt
     6001 tttttgatga ctttgcgatt ttgttgttgc tttgcagtaa attgcaagat ttaataaaaa
     6061 aacgcaaagc aatgattaaa ggatgttcag aatgaaactc atggaaacac ttaaccagtg
     6121 cataaacgct ggtcatgaaa tgacgaaggc tatcgccatt gcacagttta atgatgacag
     6181 cccggaagcg aggaaaataa cccggcgctg gagaataggt gaagcagcgg atttagttgg
     6241 ggtttcttct caggctatca gagatgccga gaaagcaggg cgactaccgc acccggatat
     6301 ggaaattcga ggacgggttg agcaacgtgt tggttataca attgaacaaa ttaatcatat
     6361 gcgtgatgtg tttggtacgc gattgcgacg tgctgaagac gtatttccac cggtgatcgg
     6421 ggttgctgcc cataaaggtg gcgtttacaa aacctcagtt tctgttcatc ttgctcagga
     6481 tctggctctg aaggggctac gtgttttgct cgtggaaggt aacgaccccc agggaacagc
     6541 ctcaatgtat cacggatggg taccagatct tcatattcat gcagaagaca ctctcctgcc
     6601 tttctatctt ggggaaaagg acgatgtcac ttatgcaata aagcccactt gctggccggg
     6661 gcttgacatt attccttcct gtctggctct gcaccgtatt gaaactgagt taatgggcaa
     6721 atttgatgaa ggtaaactgc ccaccgatcc acacctgatg ctccgactgg ccattgaaac
     6781 tgttgctcat gactatgatg tcatagttat tgacagcgcg cctaacctgg gtatcggcac
     6841 gattaatgtc gtatgtgctg ctgatgtgct gattgttccc acgcctgctg agttgtttga
     6901 ctacacctcc gcactgcagt ttttcgatat gcttcgtgat ctgctcaaga acgttgatct
     6961 taaagggttc gagcctgatg tacgtatttt gcttaccaaa tacagcaata gtaatggctc
     7021 tcagtccccg tggatggagg agcaaattcg ggatgcctgg ggaagcatgg ttctaaaaaa
     7081 tgttgtacgt gaaacggatg aagttggtaa aggtcagatc cggatgagaa ctgtttttga
     7141 acaggccatt gatcaacgct cttcaactgg tgcctggaga aatgctcttt ctatttggga
     7201 acctgtctgc aatgaaattt tcgatcgtct gattaaacca cgctgggaga ttagataatg
     7261 aagcgtgcgc ctgttattcc aaaacatacg ctcaatactc aaccggttga agatacttcg
     7321 ttatcgacac cagctgcccc gatggtggat tcgttaattg cgcgcgtagg agtaatggct
     7381 cgcggtaatg ccattacttt gcctgtatgt ggtcgggatg tgaagtttac tcttgaagtg
     7441 ctccggggtg atagtgttga gaagacctct cgggtatggt caggtaatga acgtgaccag
     7501 gagctgctta ctgaggacgc actggatgat ctcatccctt cttttctact gactggtcaa
     7561 cagacaccgg cgttcggtcg aagagtatct ggtgtcatag aaattgccga tgggagtcgc
     7621 cgtcgtaaag ctgctgcact taccgaaagt gattatcgtg ttctggttgg cgagctggat
     7681 gatgagcaga tggctgcatt atccagattg ggtaacgatt atcgcccaac aagtgcttat
     7741 gaacgtggtc agcgttatgc aagccgattg cagaatgaat ttgctggaaa tatttctgcg
     7801 ctggctgatg cggaaaatat ttcacgtaag attattaccc gctgtatcaa caccgccaaa
     7861 ttgcctaaat cagttgttgc tcttttttct caccccggtg aactatctgc ccggtcaggt
     7921 gatgcacttc aaaaagcctt tacagataaa gaggaattac ttaagcagca ggcatctaac
     7981 cttcatgagc agaaaaaagc tggggtgata tttgaagctg aagaagttat cactctttta
     8041 acttctgtgc ttaaaacgtc atctgcatca agaactagtt taagctcacg acatcagttt
     8101 gctcctggag cgacagtatt gtataagggc gataaaatgg tgcttaacct ggacaggtcc
     8161 cgtgttccaa ctgagtgtat agagaaaatt gaggccattc ttaaggaact tgaaaagcca
     8221 gcaccctgat gcgaccacgt tttagtctac gtttatctgt ctttacttaa tgtcctttgt
     8281 tacaggccag aaagcataac tggcctgaat attctctctg ggcccactgt tccacttgta
     8341 tcgtcggtct gataatcaga ctgggaccac ggtcccactc gtatcgtcgg tctgattatt
     8401 agtctgggac cacggtccca ctcgtatcgt cggtctgatt attagtctgg gaccacggtc
     8461 ccactcgtat cgtcggtctg ataatcagac tgggaccacg gtcccactcg tatcgtcggt
     8521 ctgattatta gtctgggacc atggtcccac tcgtatcgtc ggtctgatta ttagtctggg
     8581 accacggtcc cactcgtatc gtcggtctga ttattagtct ggaaccacgg tcccactcgt
     8641 atcgtcggtc tgattattag tctgggacca cggtcccact cgtatcgtcg gtctgattat
     8701 tagtctggga ccacgatccc actcgtgttg tcggtctgat tatcggtctg ggaccacggt
     8761 cccacttgta ttgtcgatca gactatcagc gtgagactac gattccatca atgcctgtca
     8821 agggcaagta ttgacatgtg tcgtggtttg tctggtcaac caccgcggtc tcagtggtgt
     8881 acggtacaaa ccccgacgtg cacagctgtc accggatgtg ctttccggtc tgatgagtcc
     8941 gtgaggacga aacagcctct acaaataatt ttgtttaata gtttacgagg taaggaggtt
     9001 taag
//
