LOCUS       sequence-178109-        7954 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 7954)
  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 7954)
  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: TP901 GFP BAC Reporter
FEATURES             Location/Qualifiers
     source          1..7954
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        275..324
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             360..368
                     /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(404..432)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1027..1047
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1150..1236
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1481..1499)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(1542..2201)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2116..2135
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2202..2304)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      3162..3776
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      3253..3784
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      3852..4071
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             4162..4917
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     CDS             5587..6762
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             6762..7733
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
ORIGIN
        1 accacggtcc cactcgtatc gtcggtctga ttattagtct gggaccacgg tcccactcgt
       61 atcgtcggtc tgattattag tctgggacca cgatcccact cgtgttgtcg gtctgattat
      121 cggtctggga ccacggtccc acttgtattg tcgatcagac tatcagcgtg agactacgat
      181 tccatcaatg cctgtcaagg gcaagtacgg tgctcggcga gtttttattt cgtttatttc
      241 aattaaggta actaaaaaac tcctttaaac acagctgtca ccggatgtgc tttccggtct
      301 gatgagtccg tgaggacgaa acagcctcta caaataattt tgtttaatag tttacgaggt
      361 aaggaggttt aagatgagta aaggagaaga acttttcact ggagttgtcc caattcttgt
      421 tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga
      481 tgcaacatac ggaaaactta cccttaaatt tatttgcact actggaaaac tacctgttcc
      541 atggccaaca cttgtcacta ctttcggtta tggtgttcaa tgctttgcga gatacccaga
      601 tcatatgaaa cagcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaaag
      661 aactatattt ttcaaagatg acgggaacta caagacacgt gctgaagtca agtttgaagg
      721 tgataccctt gttaatagaa tcgagttaaa aggtattgat tttaaagaag atggaaacat
      781 tcttggacac aaattggaat acaactataa ctcacacaat gtatacatca tggcagacaa
      841 acaaaagaat ggaatcaaag ttaacttcaa aattagacac aacattgaag atggaagcgt
      901 tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc
      961 agacaaccat tacctgtcca cacaatctgc cctttcgaaa gatcccaacg aaaagagaga
     1021 ccacatggtc cttcttgagt ttgtaacagc tgctgggatt acacatggca tggatgatct
     1081 ctacaaataa aagcttgata tcgaattcct gcagcccggg ggatcccatg gtacgcgtgc
     1141 tagaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     1201 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgccctagaa tcgatgcaaa
     1261 aaaagcaaaa agcatttacc ttgattgaga tgttaattgt gttggcattt aattaaaaag
     1321 ttaaacaaaa ttatttgtag agggaaaccg ttgtggtctc cctgaatata ttatacgagc
     1381 cttatgcatg cccgtaaagt tatccagcaa ccactcatag acctagggca gcagataggg
     1441 acgacgtggt gttagctgtg ctcgaggtga agacgaaagg gcctcgtgat acgcctattt
     1501 ttataggtta atgtcatgat aataatggtt tcttagacgt cttacgcccc gccctgccac
     1561 tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg
     1621 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     1681 cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg
     1741 gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg
     1801 aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc
     1861 cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa
     1921 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata
     1981 cgaaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac
     2041 ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg
     2101 ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg
     2161 gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct
     2221 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
     2281 ttggaacctc ttacgtgccg atcaacgtct cattttcgcc aaaagttggc ccagggcttc
     2341 ccggtatcaa cagggacacc aggatttatt tattctgcga agtgatcttc cgtcacaggt
     2401 atttattcgc gataagctca tggagcggcg taaccgtcgc acaggaagga cagagaaagc
     2461 gcggatctgg gaagtgacgg acagaacggt caggacctgg attggggagg cggttgccgc
     2521 cgctgctgct gacggtgtga cgttctctgt tccggtcaca ccacatacgt tccgccattc
     2581 ctatgcgatg cacatgctgt atgccggtat accgctgaaa gttctgcaaa gcctgatggg
     2641 acataagtcc atcagttcaa cggaagtcta cacgaaggtt tttgcgctgg atgtggctgc
     2701 ccggcaccgg gtgcagtttg cgatgccgga gtctgatgcg gttgcgatgc tgaaacaatt
     2761 atcctgagaa taaatgcctt ggcctttata tggaaatgtg gaactgagtg gatatgctgt
     2821 ttttgtctgt taaacagaga agctggctgt tatccactga gaagcgaacg aaacagtcgg
     2881 gaaaatctcc cattatcgta gagatccgca ttattaatct caggagcctg tgtagcgttt
     2941 ataggaagta gtgttctgtc atgatgcctg caagcggtaa cgaaaacgat ttgaatatgc
     3001 cttcaggaac aatagaaatc ttcgtgcggt gttacgttga agtggagcgg attatgtcag
     3061 caatggacag aacaacctaa tgaacacaga accatgatgt ggtctgtcct tttacagcca
     3121 gtagtgctcg ccgcagtcga gcgacagggc gaagccctcg gctggttgcc ctcgccgctg
     3181 ggctggcggc cgtctatggc cctgcaaacg cgccagaaac gccgtcgaag ccgtgtgcga
     3241 gacaccgcgg ccggccgccg gcgttgtgga tacctcgcgg aaaacttggc cctcactgac
     3301 agatgagggg cggacgttga cacttgaggg gccgactcac ccggcgcggc gttgacagat
     3361 gaggggcagg ctcgatttcg gccggcgacg tggagctggc cagcctcgca aatcggcgaa
     3421 aacgcctgat tttacgcgag tttcccacag atgatgtgga caagcctggg gataagtgcc
     3481 ctgcggtatt gacacttgag gggcgcgact actgacagat gaggggcgcg atccttgaca
     3541 cttgaggggc agagtgctga cagatgaggg gcgcacctat tgacatttga ggggctgtcc
     3601 acaggcagaa aatccagcat ttgcaagggt ttccgcccgt ttttcggcca ccgctaacct
     3661 gtcttttaac ctgcttttaa accaatattt ataaaccttg tttttaacca gggctgcgcc
     3721 ctgtgcgcgt gaccgcgcac gccgaagggg ggtgcccccc cttctcgaac cctcccggtc
     3781 gagtgagcga ggaagcacca gggaacagca cttatatatt ctgcttacac acgatgcctg
     3841 aaaaaacttc ccttggggtt atccacttat ccacggggat atttttataa ttattttttt
     3901 tatagttttt agatcttctt ttttagagcg ccttgtaggc ctttatccat gctggttcta
     3961 gagaaggtgt tgtgacaaat tgccctttca gtgtgacaaa tcaccctcaa atgacagtcc
     4021 tgtctgtgac aaattgccct taaccctgtg acaaattgcc ctcagaagaa gctgtttttt
     4081 cacaaagtta tccctgctta ttgactcttt tttatttagt gtgacaatct aaaaacttgt
     4141 cacacttcac atggatctgt catggcggaa acagcggtta tcaatcacaa gaaacgtaaa
     4201 aatagcccgc gaatcgtcca gtcaaacgac ctcactgagg cggcatatag tctctcccgg
     4261 gatcaaaaac gtatgctgta tctgttcgtt gaccagatca gaaaatctga tggcacccta
     4321 caggaacatg acggtatctg cgagatccat gttgctaaat atgctgaaat attcggattg
     4381 acctctgcgg aagccagtaa ggatatacgg caggcattga agagtttcgc ggggaaggaa
     4441 gtggtttttt atcgccctga agaggatgcc ggcgatgaaa aaggctatga atcttttcct
     4501 tggtttatca aacgtgcgca cagtccatcc agagggcttt acagtgtaca tatcaaccca
     4561 tatctcattc ccttctttat cgggttacag aaccggttta cgcagtttcg gcttagtgaa
     4621 acaaaagaaa tcaccaatcc gtatgccatg cgtttatacg aatccctgtg tcagtatcgt
     4681 aagccggatg gctcaggcat cgtctctctg aaaatcgact ggatcataga gcgttaccag
     4741 ctgcctcaaa gttaccagcg tatgcctgac ttccgccgcc gcttcctgca ggtctgtgtt
     4801 aatgagatca acagcagaac tccaatgcgc ctctcataca ttgagaaaaa gaaaggccgc
     4861 cagacgactc atatcgtatt ttccttccgc gatatcactt ccatgacgac aggatagtct
     4921 gagggttatc tgtcacagat ttgagggtgg ttcgtcacat ttgttctgac ctactgaggg
     4981 taatttgtca cagttttgct gtttccttca gcctgcatgg attttctcat actttttgaa
     5041 ctgtaatttt taaggaagcc aaatttgagg gtaatttgtc acagttttgc tgtttccttc
     5101 agcctgcatg gattttctca tactttttga actgtaattt ttaaggaagc caaatttgag
     5161 ggcagtttgt cacagttgat ttccttctct ttcccttcgt catgtgactt gatatcgggg
     5221 gttagttcgt catcattgat gagggttgat tatcacagtt tattactctg aattggctat
     5281 ccgcgtgtgt acctctacct ggagtttttc ccacggtgga tatttcttct tgcgctgagc
     5341 gtaagagcta tctgacagaa cagttcttct ttgcttcctc gccagttcgc tcgctatgct
     5401 cggttacacg gctgcggcga gcgctagtga taataagtga ctgaggtatg tgctcttctt
     5461 atctcctttt gtagtgttgc tcttatttta aacaactttg cggttttttg atgactttgc
     5521 gattttgttg ttgctttgca gtaaattgca agatttaata aaaaaacgca aagcaatgat
     5581 taaaggatgt tcagaatgaa actcatggaa acacttaacc agtgcataaa cgctggtcat
     5641 gaaatgacga aggctatcgc cattgcacag tttaatgatg acagcccgga agcgaggaaa
     5701 ataacccggc gctggagaat aggtgaagca gcggatttag ttggggtttc ttctcaggct
     5761 atcagagatg ccgagaaagc agggcgacta ccgcacccgg atatggaaat tcgaggacgg
     5821 gttgagcaac gtgttggtta tacaattgaa caaattaatc atatgcgtga tgtgtttggt
     5881 acgcgattgc gacgtgctga agacgtattt ccaccggtga tcggggttgc tgcccataaa
     5941 ggtggcgttt acaaaacctc agtttctgtt catcttgctc aggatctggc tctgaagggg
     6001 ctacgtgttt tgctcgtgga aggtaacgac ccccagggaa cagcctcaat gtatcacgga
     6061 tgggtaccag atcttcatat tcatgcagaa gacactctcc tgcctttcta tcttggggaa
     6121 aaggacgatg tcacttatgc aataaagccc acttgctggc cggggcttga cattattcct
     6181 tcctgtctgg ctctgcaccg tattgaaact gagttaatgg gcaaatttga tgaaggtaaa
     6241 ctgcccaccg atccacacct gatgctccga ctggccattg aaactgttgc tcatgactat
     6301 gatgtcatag ttattgacag cgcgcctaac ctgggtatcg gcacgattaa tgtcgtatgt
     6361 gctgctgatg tgctgattgt tcccacgcct gctgagttgt ttgactacac ctccgcactg
     6421 cagtttttcg atatgcttcg tgatctgctc aagaacgttg atcttaaagg gttcgagcct
     6481 gatgtacgta ttttgcttac caaatacagc aatagtaatg gctctcagtc cccgtggatg
     6541 gaggagcaaa ttcgggatgc ctggggaagc atggttctaa aaaatgttgt acgtgaaacg
     6601 gatgaagttg gtaaaggtca gatccggatg agaactgttt ttgaacaggc cattgatcaa
     6661 cgctcttcaa ctggtgcctg gagaaatgct ctttctattt gggaacctgt ctgcaatgaa
     6721 attttcgatc gtctgattaa accacgctgg gagattagat aatgaagcgt gcgcctgtta
     6781 ttccaaaaca tacgctcaat actcaaccgg ttgaagatac ttcgttatcg acaccagctg
     6841 ccccgatggt ggattcgtta attgcgcgcg taggagtaat ggctcgcggt aatgccatta
     6901 ctttgcctgt atgtggtcgg gatgtgaagt ttactcttga agtgctccgg ggtgatagtg
     6961 ttgagaagac ctctcgggta tggtcaggta atgaacgtga ccaggagctg cttactgagg
     7021 acgcactgga tgatctcatc ccttcttttc tactgactgg tcaacagaca ccggcgttcg
     7081 gtcgaagagt atctggtgtc atagaaattg ccgatgggag tcgccgtcgt aaagctgctg
     7141 cacttaccga aagtgattat cgtgttctgg ttggcgagct ggatgatgag cagatggctg
     7201 cattatccag attgggtaac gattatcgcc caacaagtgc ttatgaacgt ggtcagcgtt
     7261 atgcaagccg attgcagaat gaatttgctg gaaatatttc tgcgctggct gatgcggaaa
     7321 atatttcacg taagattatt acccgctgta tcaacaccgc caaattgcct aaatcagttg
     7381 ttgctctttt ttctcacccc ggtgaactat ctgcccggtc aggtgatgca cttcaaaaag
     7441 cctttacaga taaagaggaa ttacttaagc agcaggcatc taaccttcat gagcagaaaa
     7501 aagctggggt gatatttgaa gctgaagaag ttatcactct tttaacttct gtgcttaaaa
     7561 cgtcatctgc atcaagaact agtttaagct cacgacatca gtttgctcct ggagcgacag
     7621 tattgtataa gggcgataaa atggtgctta acctggacag gtcccgtgtt ccaactgagt
     7681 gtatagagaa aattgaggcc attcttaagg aacttgaaaa gccagcaccc tgatgcgacc
     7741 acgttttagt ctacgtttat ctgtctttac ttaatgtcct ttgttacagg ccagaaagca
     7801 taactggcct gaatattctc tctgggccca ctgttccact tgtatcgtcg gtctgataat
     7861 cagactggga ccacggtccc actcgtatcg tcggtctgat tattagtctg ggaccacggt
     7921 cccactcgta tcgtcggtct gattattagt ctgg
//
