LOCUS       sequence-148754-        7878 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Output plasmid: contains 5-state register made up of TP901 and BxbI 
            recognition sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7878)
  AUTHORS   Roquet N, Soleimany AP, Ferris AC, Aaronson S, Lu TK
  TITLE     Synthetic recombinase-based state machines in living cells.
  JOURNAL   Science. 2016 Jul 22;353(6297):aad8559. doi: 
            10.1126/science.aad8559. Epub 2016 Jul 21.
  PUBMED    27463678
REFERENCE   2  (bases 1 to 7878)
  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: "Science."; 
            date: "2016-07-22"; volume: "353(6297):aad8559. doi"; pages: " 
            10.1126/science.aad8559. Epub 2016 Jul 21"
COMMENT     Sequence Label: pNR160
FEATURES             Location/Qualifiers
     source          1..7878
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      340..559
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      627..1158
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      635..1249
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     promoter        2107..2209
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2210..2869
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2276..2295)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    3383..3431
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     protein_bind    3567..3624
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attP site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     misc_feature    4101..4574
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     CDS             complement(4647..5618)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(5618..6793)
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     misc_feature    7119..7369
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(join(7372..7878,1..249))
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
ORIGIN
        1 ccgtatatcc ttactggctt ccgcagaggt caatccgaat atttcagcat atttagcaac
       61 atggatctcg cagataccgt catgttcctg tagggtgcca tcagattttc tgatctggtc
      121 aacgaacaga tacagcatac gtttttgatc ccgggagaga ctatatgccg cctcagtgag
      181 gtcgtttgac tggacgattc gcgggctatt tttacgtttc ttgtgattga taaccgctgt
      241 ttccgccatg acagatccat gtgaagtgtg acaagttttt agattgtcac actaaataaa
      301 aaagagtcaa taagcaggga taactttgtg aaaaaacagc ttcttctgag ggcaatttgt
      361 cacagggtta agggcaattt gtcacagaca ggactgtcat ttgagggtga tttgtcacac
      421 tgaaagggca atttgtcaca acaccttctc tagaaccagc atggataaag gcctacaagg
      481 cgctctaaaa aagaagatct aaaaactata aaaaaaataa ttataaaaat atccccgtgg
      541 ataagtggat aaccccaagg gaagtttttt caggcatcgt gtgtaagcag aatatataag
      601 tgctgttccc tggtgcttcc tcgctcactc gaccgggagg gttcgagaag ggggggcacc
      661 ccccttcggc gtgcgcggtc acgcgcacag ggcgcagccc tggttaaaaa caaggtttat
      721 aaatattggt ttaaaagcag gttaaaagac aggttagcgg tggccgaaaa acgggcggaa
      781 acccttgcaa atgctggatt ttctgcctgt ggacagcccc tcaaatgtca ataggtgcgc
      841 ccctcatctg tcagcactct gcccctcaag tgtcaaggat cgcgcccctc atctgtcagt
      901 agtcgcgccc ctcaagtgtc aataccgcag ggcacttatc cccaggcttg tccacatcat
      961 ctgtgggaaa ctcgcgtaaa atcaggcgtt ttcgccgatt tgcgaggctg gccagctcca
     1021 cgtcgccggc cgaaatcgag cctgcccctc atctgtcaac gccgcgccgg gtgagtcggc
     1081 ccctcaagtg tcaacgtccg cccctcatct gtcagtgagg gccaagtttt ccgcgaggta
     1141 tccacaacgc cggcggccgg ccgcggtgtc tcgcacacgg cttcgacggc gtttctggcg
     1201 cgtttgcagg gccatagacg gccgccagcc cagcggcgag ggcaaccagc cgagggcttc
     1261 gccctgtcgc tcgactgcgg cgagcactac tggctgtaaa aggacagacc acatcatggt
     1321 tctgtgttca ttaggttgtt ctgtccattg ctgacataat ccgctccact tcaacgtaac
     1381 accgcacgaa gatttctatt gttcctgaag gcatattcaa atcgttttcg ttaccgcttg
     1441 caggcatcat gacagaacac tacttcctat aaacgctaca caggctcctg agattaataa
     1501 tgcggatctc tacgataatg ggagattttc ccgactgttt cgttcgcttc tcagtggata
     1561 acagccagct tctctgttta acagacaaaa acagcatatc cactcagttc cacatttcca
     1621 tataaaggcc aaggcattta ttctcaggat aattgtttca gcatcgcaac cgcatcagac
     1681 tccggcatcg caaactgcac ccggtgccgg gcagccacat ccagcgcaaa aaccttcgtg
     1741 tagacttccg ttgaactgat ggacttatgt cccatcaggc tttgcagaac tttcagcggt
     1801 ataccggcat acagcatgtg catcgcatag gaatggcgga acgtatgtgg tgtgaccgga
     1861 acagagaacg tcacaccgtc agcagcagcg gcggcaaccg cctccccaat ccaggtcctg
     1921 accgttctgt ccgtcacttc ccagatccgc gctttctctg tccttcctgt gcgacggtta
     1981 cgccgctcca tgagcttatc gcgaataaat acctgtgacg gaagatcact tcgcagaata
     2041 aataaatcct ggtgtccctg ttgataccgg gaagccctgg gccaactttt ggcgaaaatg
     2101 agacgttgat cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc
     2161 gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa
     2221 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
     2281 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
     2341 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
     2401 ccgcctgatg aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat
     2461 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
     2521 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
     2581 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     2641 cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     2701 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     2761 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
     2821 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaag acgtctaagt
     2881 aagtgcccat tcagccagta taaccagata aaaaaaatcc ttagctttcg ctaaggatga
     2941 tttctgtcag ccaggaaggt ataacaggaa aaattaaagg ggagataaaa tccccccttt
     3001 ttggttaact cgaggattcg aaaaggtgaa atctcctttc acggcccatt cctcatggat
     3061 gggccgttta tttccccgag ccagacgtcc accatacctt cgatattcgc gcccactctc
     3121 ccattagtcg gagtggatgt gttgcgattg cccgctagat caatgtgcac gtcgagtgcg
     3181 gtattcctct gggcatgcca acacaattaa catctcaatc aaggtaaatg ctttttgctt
     3241 tttttgccgt ggtgcggatc tagctacccg cacttgagtc cacggctgat gccaacacaa
     3301 ttaacatcag aatcaaggta aatgcttttt gctttttttg cgggctcact agggaatggc
     3361 gtcgtccgtg acattctgtg cgcggccggc ttgtcgacga cggcggtctc cgtcgtcagg
     3421 atcatccggg ccagcggctg tctatgccag aagtcggtca ggtcggagtc ctagaaagga
     3481 gttttttagt taccttaatt ctaataaacg aaataaaaac tcgccgttcg cgcgacatga
     3541 catgccgaca caatctccct gctcaggtcg tggtttgtct ggtcaaccac cgcggtctca
     3601 gtggtgtacg gtacaaaccc cgacgcaata gcgccaacca tgcaccatag ccagcctgac
     3661 agtagccaaa ggagtttttt agttacctta attgaaataa acgaaataaa aactcgcgca
     3721 gctgagagcg acacagttgc ctcactccta gggtgtaata ggcactaaaa gagtgatatg
     3781 gcacgcgcat gtggaacgcc gttagagggt tgctgagcca tcgtggtcct tagactcggg
     3841 aaaaacaaaa acccgccgaa gcgggttaag tgcggggcgg ctagcccgcc acctaagtta
     3901 tgagtcagga aaaaaggcga cagagtaatc tgtcgccttt tttctttgct tgctttgtaa
     3961 tcactcgatg accaaaagta aaaacccgcc gaagcgggtt tttacgtaaa acaggtgaaa
     4021 ctgacgctag ccggtcacat gtcaatactt gcccttgaca ggcattgatg gaatcgtagt
     4081 ctcacgctga tagtctgatc gacaatacaa gtgggaccgt ggtcccagac cgataatcag
     4141 accgacaaca cgagtgggat cgtggtccca gactaataat cagaccgacg atacgagtgg
     4201 gaccgtggtc ccagactaat aatcagaccg acgatacgag tgggaccgtg gttccagact
     4261 aataatcaga ccgacgatac gagtgggacc gtggtcccag actaataatc agaccgacga
     4321 tacgagtggg accatggtcc cagactaata atcagaccga cgatacgagt gggaccgtgg
     4381 tcccagtctg attatcagac cgacgatacg agtgggaccg tggtcccaga ctaataatca
     4441 gaccgacgat acgagtggga ccgtggtccc agactaataa tcagaccgac gatacgagtg
     4501 ggaccgtggt cccagtctga ttatcagacc gacgatacaa gtggaacagt gggcccagag
     4561 agaatattca ggccagttat gctttctggc ctgtaacaaa ggacattaag taaagacaga
     4621 taaacgtaga ctaaaacgtg gtcgcatcag ggtgctggct tttcaagttc cttaagaatg
     4681 gcctcaattt tctctataca ctcagttgga acacgggacc tgtccaggtt aagcaccatt
     4741 ttatcgccct tatacaatac tgtcgctcca ggagcaaact gatgtcgtga gcttaaacta
     4801 gttcttgatg cagatgacgt tttaagcaca gaagttaaaa gagtgataac ttcttcagct
     4861 tcaaatatca ccccagcttt tttctgctca tgaaggttag atgcctgctg cttaagtaat
     4921 tcctctttat ctgtaaaggc tttttgaagt gcatcacctg accgggcaga tagttcaccg
     4981 gggtgagaaa aaagagcaac aactgattta ggcaatttgg cggtgttgat acagcgggta
     5041 ataatcttac gtgaaatatt ttccgcatca gccagcgcag aaatatttcc agcaaattca
     5101 ttctgcaatc ggcttgcata acgctgacca cgttcataag cacttgttgg gcgataatcg
     5161 ttacccaatc tggataatgc agccatctgc tcatcatcca gctcgccaac cagaacacga
     5221 taatcacttt cggtaagtgc agcagcttta cgacggcgac tcccatcggc aatttctatg
     5281 acaccagata ctcttcgacc gaacgccggt gtctgttgac cagtcagtag aaaagaaggg
     5341 atgagatcat ccagtgcgtc ctcagtaagc agctcctggt cacgttcatt acctgaccat
     5401 acccgagagg tcttctcaac actatcaccc cggagcactt caagagtaaa cttcacatcc
     5461 cgaccacata caggcaaagt aatggcatta ccgcgagcca ttactcctac gcgcgcaatt
     5521 aacgaatcca ccatcggggc agctggtgtc gataacgaag tatcttcaac cggttgagta
     5581 ttgagcgtat gttttggaat aacaggcgca cgcttcatta tctaatctcc cagcgtggtt
     5641 taatcagacg atcgaaaatt tcattgcaga caggttccca aatagaaaga gcatttctcc
     5701 aggcaccagt tgaagagcgt tgatcaatgg cctgttcaaa aacagttctc atccggatct
     5761 gacctttacc aacttcatcc gtttcacgta caacattttt tagaaccatg cttccccagg
     5821 catcccgaat ttgctcctcc atccacgggg actgagagcc attactattg ctgtatttgg
     5881 taagcaaaat acgtacatca ggctcgaacc ctttaagatc aacgttcttg agcagatcac
     5941 gaagcatatc gaaaaactgc agtgcggagg tgtagtcaaa caactcagca ggcgtgggaa
     6001 caatcagcac atcagcagca catacgacat taatcgtgcc gatacccagg ttaggcgcgc
     6061 tgtcaataac tatgacatca tagtcatgag caacagtttc aatggccagt cggagcatca
     6121 ggtgtggatc ggtgggcagt ttaccttcat caaatttgcc cattaactca gtttcaatac
     6181 ggtgcagagc cagacaggaa ggaataatgt caagccccgg ccagcaagtg ggctttattg
     6241 cataagtgac atcgtccttt tccccaagat agaaaggcag gagagtgtct tctgcatgaa
     6301 tatgaagatc tggtacccat ccgtgataca ttgaggctgt tccctggggg tcgttacctt
     6361 ccacgagcaa aacacgtagc cccttcagag ccagatcctg agcaagatga acagaaactg
     6421 aggttttgta aacgccacct ttatgggcag caaccccgat caccggtgga aatacgtctt
     6481 cagcacgtcg caatcgcgta ccaaacacat cacgcatatg attaatttgt tcaattgtat
     6541 aaccaacacg ttgctcaacc cgtcctcgaa tttccatatc cgggtgcggt agtcgccctg
     6601 ctttctcggc atctctgata gcctgagaag aaaccccaac taaatccgct gcttcaccta
     6661 ttctccagcg ccgggttatt ttcctcgctt ccgggctgtc atcattaaac tgtgcaatgg
     6721 cgatagcctt cgtcatttca tgaccagcgt ttatgcactg gttaagtgtt tccatgagtt
     6781 tcattctgaa catcctttaa tcattgcttt gcgttttttt attaaatctt gcaatttact
     6841 gcaaagcaac aacaaaatcg caaagtcatc aaaaaaccgc aaagttgttt aaaataagag
     6901 caacactaca aaaggagata agaagagcac atacctcagt cacttattat cactagcgct
     6961 cgccgcagcc gtgtaaccga gcatagcgag cgaactggcg aggaagcaaa gaagaactgt
     7021 tctgtcagat agctcttacg ctcagcgcaa gaagaaatat ccaccgtggg aaaaactcca
     7081 ggtagaggta cacacgcgga tagccaattc agagtaataa actgtgataa tcaaccctca
     7141 tcaatgatga cgaactaacc cccgatatca ggtcacatga cgaagggaaa gagaaggaaa
     7201 tcaactgtga caaactgccc tcaaatttgg cttccttaaa aattacagtt caaaaagtat
     7261 gagaaaatcc atgcaggctg aaggaaacag caaaactgtg acaaattacc ctcagtaggt
     7321 cagaacaaat gtgacgaacc accctcaaat ctgtgacaga taaccctcag actatcctgt
     7381 cgtcatggaa gtgatatcgc ggaaggaaaa tacgatatga gtcgtctggc ggcctttctt
     7441 tttctcaatg tatgagaggc gcattggagt tctgctgttg atctcattaa cacagacctg
     7501 caggaagcgg cggcggaagt caggcatacg ctggtaactt tgaggcagct ggtaacgctc
     7561 tatgatccag tcgattttca gagagacgat gcctgagcca tccggcttac gatactgaca
     7621 cagggattcg tataaacgca tggcatacgg attggtgatt tcttttgttt cactaagccg
     7681 aaactgcgta aaccggttct gtaacccgat aaagaaggga atgagatatg ggttgatatg
     7741 tacactgtaa agccctctgg atggactgtg cgcacgtttg ataaaccaag gaaaagattc
     7801 atagcctttt tcatcgccgg catcctcttc agggcgataa aaaaccactt ccttccccgc
     7861 gaaactcttc aatgcctg
//
