LOCUS       sequence-178111-        7977 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Synthetic gene circuit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7977)
  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 7977)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: PhiC31 GFP BAC Reporter
FEATURES             Location/Qualifiers
     source          1..7977
                     /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    889..922
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     primer_bind     complement(1100..1118)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(1161..1820)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1735..1754
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1821..1923)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      2781..3395
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      2872..3403
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      3471..3690
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             3781..4536
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    4539..4789
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             5115..6290
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             6290..7261
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_RNA        7879..7928
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             7964..7972
                     /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 atggagtacg cgcccgggga
      901 gcccaagggc acgccctggc acccgcatta attaaaaagt taaacaaaat tatttgtaga
      961 gggaaaccgt tgtggtctcc ctgaatatat tatacgagcc ttatgcatgc ccgtaaagtt
     1021 atccagcaac cactcataga cctagggcag cagataggga cgacgtggtg ttagctgtgc
     1081 tcgaggtgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     1141 ataatggttt cttagacgtc ttacgccccg ccctgccact catcgcagta ctgttgtaat
     1201 tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc
     1261 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg
     1321 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg
     1381 gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg
     1441 taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca
     1501 ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca
     1561 ctatcccata tcaccagctc accgtctttc attgccatac gaaattccgg atgagcattc
     1621 atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg
     1681 gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact
     1741 gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat
     1801 ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa
     1861 aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga
     1921 tcaacgtctc attttcgcca aaagttggcc cagggcttcc cggtatcaac agggacacca
     1981 ggatttattt attctgcgaa gtgatcttcc gtcacaggta tttattcgcg ataagctcat
     2041 ggagcggcgt aaccgtcgca caggaaggac agagaaagcg cggatctggg aagtgacgga
     2101 cagaacggtc aggacctgga ttggggaggc ggttgccgcc gctgctgctg acggtgtgac
     2161 gttctctgtt ccggtcacac cacatacgtt ccgccattcc tatgcgatgc acatgctgta
     2221 tgccggtata ccgctgaaag ttctgcaaag cctgatggga cataagtcca tcagttcaac
     2281 ggaagtctac acgaaggttt ttgcgctgga tgtggctgcc cggcaccggg tgcagtttgc
     2341 gatgccggag tctgatgcgg ttgcgatgct gaaacaatta tcctgagaat aaatgccttg
     2401 gcctttatat ggaaatgtgg aactgagtgg atatgctgtt tttgtctgtt aaacagagaa
     2461 gctggctgtt atccactgag aagcgaacga aacagtcggg aaaatctccc attatcgtag
     2521 agatccgcat tattaatctc aggagcctgt gtagcgttta taggaagtag tgttctgtca
     2581 tgatgcctgc aagcggtaac gaaaacgatt tgaatatgcc ttcaggaaca atagaaatct
     2641 tcgtgcggtg ttacgttgaa gtggagcgga ttatgtcagc aatggacaga acaacctaat
     2701 gaacacagaa ccatgatgtg gtctgtcctt ttacagccag tagtgctcgc cgcagtcgag
     2761 cgacagggcg aagccctcgg ctggttgccc tcgccgctgg gctggcggcc gtctatggcc
     2821 ctgcaaacgc gccagaaacg ccgtcgaagc cgtgtgcgag acaccgcggc cggccgccgg
     2881 cgttgtggat acctcgcgga aaacttggcc ctcactgaca gatgaggggc ggacgttgac
     2941 acttgagggg ccgactcacc cggcgcggcg ttgacagatg aggggcaggc tcgatttcgg
     3001 ccggcgacgt ggagctggcc agcctcgcaa atcggcgaaa acgcctgatt ttacgcgagt
     3061 ttcccacaga tgatgtggac aagcctgggg ataagtgccc tgcggtattg acacttgagg
     3121 ggcgcgacta ctgacagatg aggggcgcga tccttgacac ttgaggggca gagtgctgac
     3181 agatgagggg cgcacctatt gacatttgag gggctgtcca caggcagaaa atccagcatt
     3241 tgcaagggtt tccgcccgtt tttcggccac cgctaacctg tcttttaacc tgcttttaaa
     3301 ccaatattta taaaccttgt ttttaaccag ggctgcgccc tgtgcgcgtg accgcgcacg
     3361 ccgaaggggg gtgccccccc ttctcgaacc ctcccggtcg agtgagcgag gaagcaccag
     3421 ggaacagcac ttatatattc tgcttacaca cgatgcctga aaaaacttcc cttggggtta
     3481 tccacttatc cacggggata tttttataat tatttttttt atagttttta gatcttcttt
     3541 tttagagcgc cttgtaggcc tttatccatg ctggttctag agaaggtgtt gtgacaaatt
     3601 gccctttcag tgtgacaaat caccctcaaa tgacagtcct gtctgtgaca aattgccctt
     3661 aaccctgtga caaattgccc tcagaagaag ctgttttttc acaaagttat ccctgcttat
     3721 tgactctttt ttatttagtg tgacaatcta aaaacttgtc acacttcaca tggatctgtc
     3781 atggcggaaa cagcggttat caatcacaag aaacgtaaaa atagcccgcg aatcgtccag
     3841 tcaaacgacc tcactgaggc ggcatatagt ctctcccggg atcaaaaacg tatgctgtat
     3901 ctgttcgttg accagatcag aaaatctgat ggcaccctac aggaacatga cggtatctgc
     3961 gagatccatg ttgctaaata tgctgaaata ttcggattga cctctgcgga agccagtaag
     4021 gatatacggc aggcattgaa gagtttcgcg gggaaggaag tggtttttta tcgccctgaa
     4081 gaggatgccg gcgatgaaaa aggctatgaa tcttttcctt ggtttatcaa acgtgcgcac
     4141 agtccatcca gagggcttta cagtgtacat atcaacccat atctcattcc cttctttatc
     4201 gggttacaga accggtttac gcagtttcgg cttagtgaaa caaaagaaat caccaatccg
     4261 tatgccatgc gtttatacga atccctgtgt cagtatcgta agccggatgg ctcaggcatc
     4321 gtctctctga aaatcgactg gatcatagag cgttaccagc tgcctcaaag ttaccagcgt
     4381 atgcctgact tccgccgccg cttcctgcag gtctgtgtta atgagatcaa cagcagaact
     4441 ccaatgcgcc tctcatacat tgagaaaaag aaaggccgcc agacgactca tatcgtattt
     4501 tccttccgcg atatcacttc catgacgaca ggatagtctg agggttatct gtcacagatt
     4561 tgagggtggt tcgtcacatt tgttctgacc tactgagggt aatttgtcac agttttgctg
     4621 tttccttcag cctgcatgga ttttctcata ctttttgaac tgtaattttt aaggaagcca
     4681 aatttgaggg cagtttgtca cagttgattt ccttctcttt cccttcgtca tgtgacttga
     4741 tatcgggggt tagttcgtca tcattgatga gggttgatta tcacagttta ttactctgaa
     4801 ttggctatcc gcgtgtgtac ctctacctgg agtttttccc acggtggata tttcttcttg
     4861 cgctgagcgt aagagctatc tgacagaaca gttcttcttt gcttcctcgc cagttcgctc
     4921 gctatgctcg gttacacggc tgcggcgagc gctagtgata ataagtgact gaggtatgtg
     4981 ctcttcttat ctccttttgt agtgttgctc ttattttaaa caactttgcg gttttttgat
     5041 gactttgcga ttttgttgtt gctttgcagt aaattgcaag atttaataaa aaaacgcaaa
     5101 gcaatgatta aaggatgttc agaatgaaac tcatggaaac acttaaccag tgcataaacg
     5161 ctggtcatga aatgacgaag gctatcgcca ttgcacagtt taatgatgac agcccggaag
     5221 cgaggaaaat aacccggcgc tggagaatag gtgaagcagc ggatttagtt ggggtttctt
     5281 ctcaggctat cagagatgcc gagaaagcag ggcgactacc gcacccggat atggaaattc
     5341 gaggacgggt tgagcaacgt gttggttata caattgaaca aattaatcat atgcgtgatg
     5401 tgtttggtac gcgattgcga cgtgctgaag acgtatttcc accggtgatc ggggttgctg
     5461 cccataaagg tggcgtttac aaaacctcag tttctgttca tcttgctcag gatctggctc
     5521 tgaaggggct acgtgttttg ctcgtggaag gtaacgaccc ccagggaaca gcctcaatgt
     5581 atcacggatg ggtaccagat cttcatattc atgcagaaga cactctcctg cctttctatc
     5641 ttggggaaaa ggacgatgtc acttatgcaa taaagcccac ttgctggccg gggcttgaca
     5701 ttattccttc ctgtctggct ctgcaccgta ttgaaactga gttaatgggc aaatttgatg
     5761 aaggtaaact gcccaccgat ccacacctga tgctccgact ggccattgaa actgttgctc
     5821 atgactatga tgtcatagtt attgacagcg cgcctaacct gggtatcggc acgattaatg
     5881 tcgtatgtgc tgctgatgtg ctgattgttc ccacgcctgc tgagttgttt gactacacct
     5941 ccgcactgca gtttttcgat atgcttcgtg atctgctcaa gaacgttgat cttaaagggt
     6001 tcgagcctga tgtacgtatt ttgcttacca aatacagcaa tagtaatggc tctcagtccc
     6061 cgtggatgga ggagcaaatt cgggatgcct ggggaagcat ggttctaaaa aatgttgtac
     6121 gtgaaacgga tgaagttggt aaaggtcaga tccggatgag aactgttttt gaacaggcca
     6181 ttgatcaacg ctcttcaact ggtgcctgga gaaatgctct ttctatttgg gaacctgtct
     6241 gcaatgaaat tttcgatcgt ctgattaaac cacgctggga gattagataa tgaagcgtgc
     6301 gcctgttatt ccaaaacata cgctcaatac tcaaccggtt gaagatactt cgttatcgac
     6361 accagctgcc ccgatggtgg attcgttaat tgcgcgcgta ggagtaatgg ctcgcggtaa
     6421 tgccattact ttgcctgtat gtggtcggga tgtgaagttt actcttgaag tgctccgggg
     6481 tgatagtgtt gagaagacct ctcgggtatg gtcaggtaat gaacgtgacc aggagctgct
     6541 tactgaggac gcactggatg atctcatccc ttcttttcta ctgactggtc aacagacacc
     6601 ggcgttcggt cgaagagtat ctggtgtcat agaaattgcc gatgggagtc gccgtcgtaa
     6661 agctgctgca cttaccgaaa gtgattatcg tgttctggtt ggcgagctgg atgatgagca
     6721 gatggctgca ttatccagat tgggtaacga ttatcgccca acaagtgctt atgaacgtgg
     6781 tcagcgttat gcaagccgat tgcagaatga atttgctgga aatatttctg cgctggctga
     6841 tgcggaaaat atttcacgta agattattac ccgctgtatc aacaccgcca aattgcctaa
     6901 atcagttgtt gctctttttt ctcaccccgg tgaactatct gcccggtcag gtgatgcact
     6961 tcaaaaagcc tttacagata aagaggaatt acttaagcag caggcatcta accttcatga
     7021 gcagaaaaaa gctggggtga tatttgaagc tgaagaagtt atcactcttt taacttctgt
     7081 gcttaaaacg tcatctgcat caagaactag tttaagctca cgacatcagt ttgctcctgg
     7141 agcgacagta ttgtataagg gcgataaaat ggtgcttaac ctggacaggt cccgtgttcc
     7201 aactgagtgt atagagaaaa ttgaggccat tcttaaggaa cttgaaaagc cagcaccctg
     7261 atgcgaccac gttttagtct acgtttatct gtctttactt aatgtccttt gttacaggcc
     7321 agaaagcata actggcctga atattctctc tgggcccact gttccacttg tatcgtcggt
     7381 ctgataatca gactgggacc acggtcccac tcgtatcgtc ggtctgatta ttagtctggg
     7441 accacggtcc cactcgtatc gtcggtctga ttattagtct gggaccacgg tcccactcgt
     7501 atcgtcggtc tgataatcag actgggacca cggtcccact cgtatcgtcg gtctgattat
     7561 tagtctggga ccacggtccc actcgtatcg tcggtctgat tattagtctg ggaccacggt
     7621 cccactcgta tcgtcggtct gattattagt ctgggaccac ggtcccactc gtatcgtcgg
     7681 tctgattatt agtctgggac cacgatccca ctcgtgttgt cggtctgatt atcggtctgg
     7741 gaccacggtc ccacttgtat tgtcgatcag actatcagcg tgagactacg attccatcaa
     7801 tgcctgtcaa gggcaagtac ggtgctcggt gccccaactg gggtaacctt tgagttctct
     7861 cagttggggg gtgcacagct gtcaccggat gtgctttccg gtctgatgag tccgtgagga
     7921 cgaaacagcc tctacaaata attttgttta atagtttacg aggtaaggag gtttaag
//
