LOCUS       sequence-148755-        8484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Output plasmid: contains 16-state register made up of TP901, BxbI, 
            and A118 recognition sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8484)
  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 8484)
  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: pNR188
FEATURES             Location/Qualifiers
     source          1..8484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    547..604
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attP site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     protein_bind    832..880
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     protein_bind    1013..1070
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attP site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     misc_feature    1828..2301
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     CDS             complement(2374..3345)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(3345..4520)
                     /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    4846..5096
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(5099..5854)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      5945..6164
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      6232..6763
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      6240..6854
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     promoter        7712..7814
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             7815..8474
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(7881..7900)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 taagtgccca ttcagccagt ataaccagat aaaaaaaatc cttagctttc gctaaggatg
       61 atttctgtca gccaggaagg tataacagga aaaattaaag gggagataaa atcccccctt
      121 tttggttaac tcgaggattc gaaaaggtga aatctccttt cacggcccat tcctcatgga
      181 tgggccgttt atttccccga gccagacgtc caccatacct tcgatattcg cgcccactct
      241 cccattagtc ggagtggatg tgttgcgatt gcccgctaga tcaatgtgca cgacactgct
      301 cgatccgctc gcacattgcc cgctagatca atgtgtacgt cgagtgcggt attcctctgg
      361 gccggccggc ttgtcgacga cggcgcactc cgtcgtcagg atcatccggg ccgtggtgcg
      421 gatctagcta cccgcacttg agtccacggc tgatgccaac acaattaaca tcagaatcaa
      481 ggtaaatgct ttttgctttt tttgcgggct cactagggaa tggcgtcgtc cgtgacattc
      541 tgtgcggtcg gggtttgtac cgtacaccac tgagtgcgcg gtggttgacc agacaaacca
      601 cgaccagcgg ctgtctatgc cagaagtcgg tcaggtcgga gtcctaggcg agtttttatt
      661 tcgtttatta gaattaaggt aactaaaaaa ctcctttcgt tcgcgcgaca tgacatgccg
      721 acacaatctc cctgctcaga acttttcgga tcaagctatg aaaaacgcaa agagggaact
      781 aaacacttgc aatagcgcca accatgcacc atagccagcc tgacagtagc ccggccggct
      841 tgtcgacgac ggcggtctcc gtcgtcagga tcatccgggc gcagctgaga gcgacacagt
      901 cctgtacttg ggatgctggc gtttagtttc tcgtttcttc ttcttttaac gagagaaaac
      961 gaggaactaa cttggggtgc aggtaggcca tattgtcttc ggggccccac tcgtcgtggt
     1021 ttgtctggtc aaccaccgcg gtctcagtgg tgtacggtac aaaccccgac gaggcgataa
     1081 gctcggcaag gctgattacg cctgctcgtg tgatgccaac acaattaaca tctcaatcaa
     1141 ggtaaatgct ttttgctttt tttgcccgga tctccttagt gtaggttccg actcgcgctt
     1201 cgtcgacaca acttttcgga tcaagctatg aaggacgcaa agagggaact aaacacttcg
     1261 tccctgaggc tgcagttcag tacgatcgca cgccccatga aaggagtttt ttagttacct
     1321 taattgaaat aaacgaaata aaaactcgcc agatcgacag gtagaccagg tgcggtgcga
     1381 atacgagtcg tagttagttc ctcgttttct ctcgttggaa gaagaagaaa cgagaaacta
     1441 aagcaatgct ggtctggcta tgactgcctc actcctaggg tgtaataggc actaaaagag
     1501 tgatatggca cgcgcatgtg gaacgccgtt agagggttgc tgagccatcg tggtccttag
     1561 actcgggaaa aacaaaaacc cgccgaagcg ggttaagtgc ggggcggcta gcccgccacc
     1621 taagttatga gtcaggaaaa aaggcgacag agtaatctgt cgcctttttt ctttgcttgc
     1681 tttgtaatca ctcgatgacc aaaagtaaaa acccgccgaa gcgggttttt acgtaaaaca
     1741 ggtgaaactg acgctagccg gtcacatgtc aatacttgcc cttgacaggc attgatggaa
     1801 tcgtagtctc acgctgatag tctgatcgac aatacaagtg ggaccgtggt cccagaccga
     1861 taatcagacc gacaacacga gtgggatcgt ggtcccagac taataatcag accgacgata
     1921 cgagtgggac cgtggtccca gactaataat cagaccgacg atacgagtgg gaccgtggtt
     1981 ccagactaat aatcagaccg acgatacgag tgggaccgtg gtcccagact aataatcaga
     2041 ccgacgatac gagtgggacc atggtcccag actaataatc agaccgacga tacgagtggg
     2101 accgtggtcc cagtctgatt atcagaccga cgatacgagt gggaccgtgg tcccagacta
     2161 ataatcagac cgacgatacg agtgggaccg tggtcccaga ctaataatca gaccgacgat
     2221 acgagtggga ccgtggtccc agtctgatta tcagaccgac gatacaagtg gaacagtggg
     2281 cccagagaga atattcaggc cagttatgct ttctggcctg taacaaagga cattaagtaa
     2341 agacagataa acgtagacta aaacgtggtc gcatcagggt gctggctttt caagttcctt
     2401 aagaatggcc tcaattttct ctatacactc agttggaaca cgggacctgt ccaggttaag
     2461 caccatttta tcgcccttat acaatactgt cgctccagga gcaaactgat gtcgtgagct
     2521 taaactagtt cttgatgcag atgacgtttt aagcacagaa gttaaaagag tgataacttc
     2581 ttcagcttca aatatcaccc cagctttttt ctgctcatga aggttagatg cctgctgctt
     2641 aagtaattcc tctttatctg taaaggcttt ttgaagtgca tcacctgacc gggcagatag
     2701 ttcaccgggg tgagaaaaaa gagcaacaac tgatttaggc aatttggcgg tgttgataca
     2761 gcgggtaata atcttacgtg aaatattttc cgcatcagcc agcgcagaaa tatttccagc
     2821 aaattcattc tgcaatcggc ttgcataacg ctgaccacgt tcataagcac ttgttgggcg
     2881 ataatcgtta cccaatctgg ataatgcagc catctgctca tcatccagct cgccaaccag
     2941 aacacgataa tcactttcgg taagtgcagc agctttacga cggcgactcc catcggcaat
     3001 ttctatgaca ccagatactc ttcgaccgaa cgccggtgtc tgttgaccag tcagtagaaa
     3061 agaagggatg agatcatcca gtgcgtcctc agtaagcagc tcctggtcac gttcattacc
     3121 tgaccatacc cgagaggtct tctcaacact atcaccccgg agcacttcaa gagtaaactt
     3181 cacatcccga ccacatacag gcaaagtaat ggcattaccg cgagccatta ctcctacgcg
     3241 cgcaattaac gaatccacca tcggggcagc tggtgtcgat aacgaagtat cttcaaccgg
     3301 ttgagtattg agcgtatgtt ttggaataac aggcgcacgc ttcattatct aatctcccag
     3361 cgtggtttaa tcagacgatc gaaaatttca ttgcagacag gttcccaaat agaaagagca
     3421 tttctccagg caccagttga agagcgttga tcaatggcct gttcaaaaac agttctcatc
     3481 cggatctgac ctttaccaac ttcatccgtt tcacgtacaa cattttttag aaccatgctt
     3541 ccccaggcat cccgaatttg ctcctccatc cacggggact gagagccatt actattgctg
     3601 tatttggtaa gcaaaatacg tacatcaggc tcgaaccctt taagatcaac gttcttgagc
     3661 agatcacgaa gcatatcgaa aaactgcagt gcggaggtgt agtcaaacaa ctcagcaggc
     3721 gtgggaacaa tcagcacatc agcagcacat acgacattaa tcgtgccgat acccaggtta
     3781 ggcgcgctgt caataactat gacatcatag tcatgagcaa cagtttcaat ggccagtcgg
     3841 agcatcaggt gtggatcggt gggcagttta ccttcatcaa atttgcccat taactcagtt
     3901 tcaatacggt gcagagccag acaggaagga ataatgtcaa gccccggcca gcaagtgggc
     3961 tttattgcat aagtgacatc gtccttttcc ccaagataga aaggcaggag agtgtcttct
     4021 gcatgaatat gaagatctgg tacccatccg tgatacattg aggctgttcc ctgggggtcg
     4081 ttaccttcca cgagcaaaac acgtagcccc ttcagagcca gatcctgagc aagatgaaca
     4141 gaaactgagg ttttgtaaac gccaccttta tgggcagcaa ccccgatcac cggtggaaat
     4201 acgtcttcag cacgtcgcaa tcgcgtacca aacacatcac gcatatgatt aatttgttca
     4261 attgtataac caacacgttg ctcaacccgt cctcgaattt ccatatccgg gtgcggtagt
     4321 cgccctgctt tctcggcatc tctgatagcc tgagaagaaa ccccaactaa atccgctgct
     4381 tcacctattc tccagcgccg ggttattttc ctcgcttccg ggctgtcatc attaaactgt
     4441 gcaatggcga tagccttcgt catttcatga ccagcgttta tgcactggtt aagtgtttcc
     4501 atgagtttca ttctgaacat cctttaatca ttgctttgcg tttttttatt aaatcttgca
     4561 atttactgca aagcaacaac aaaatcgcaa agtcatcaaa aaaccgcaaa gttgtttaaa
     4621 ataagagcaa cactacaaaa ggagataaga agagcacata cctcagtcac ttattatcac
     4681 tagcgctcgc cgcagccgtg taaccgagca tagcgagcga actggcgagg aagcaaagaa
     4741 gaactgttct gtcagatagc tcttacgctc agcgcaagaa gaaatatcca ccgtgggaaa
     4801 aactccaggt agaggtacac acgcggatag ccaattcaga gtaataaact gtgataatca
     4861 accctcatca atgatgacga actaaccccc gatatcaggt cacatgacga agggaaagag
     4921 aaggaaatca actgtgacaa actgccctca aatttggctt ccttaaaaat tacagttcaa
     4981 aaagtatgag aaaatccatg caggctgaag gaaacagcaa aactgtgaca aattaccctc
     5041 agtaggtcag aacaaatgtg acgaaccacc ctcaaatctg tgacagataa ccctcagact
     5101 atcctgtcgt catggaagtg atatcgcgga aggaaaatac gatatgagtc gtctggcggc
     5161 ctttcttttt ctcaatgtat gagaggcgca ttggagttct gctgttgatc tcattaacac
     5221 agacctgcag gaagcggcgg cggaagtcag gcatacgctg gtaactttga ggcagctggt
     5281 aacgctctat gatccagtcg attttcagag agacgatgcc tgagccatcc ggcttacgat
     5341 actgacacag ggattcgtat aaacgcatgg catacggatt ggtgatttct tttgtttcac
     5401 taagccgaaa ctgcgtaaac cggttctgta acccgataaa gaagggaatg agatatgggt
     5461 tgatatgtac actgtaaagc cctctggatg gactgtgcgc acgtttgata aaccaaggaa
     5521 aagattcata gcctttttca tcgccggcat cctcttcagg gcgataaaaa accacttcct
     5581 tccccgcgaa actcttcaat gcctgccgta tatccttact ggcttccgca gaggtcaatc
     5641 cgaatatttc agcatattta gcaacatgga tctcgcagat accgtcatgt tcctgtaggg
     5701 tgccatcaga ttttctgatc tggtcaacga acagatacag catacgtttt tgatcccggg
     5761 agagactata tgccgcctca gtgaggtcgt ttgactggac gattcgcggg ctatttttac
     5821 gtttcttgtg attgataacc gctgtttccg ccatgacaga tccatgtgaa gtgtgacaag
     5881 tttttagatt gtcacactaa ataaaaaaga gtcaataagc agggataact ttgtgaaaaa
     5941 acagcttctt ctgagggcaa tttgtcacag ggttaagggc aatttgtcac agacaggact
     6001 gtcatttgag ggtgatttgt cacactgaaa gggcaatttg tcacaacacc ttctctagaa
     6061 ccagcatgga taaaggccta caaggcgctc taaaaaagaa gatctaaaaa ctataaaaaa
     6121 aataattata aaaatatccc cgtggataag tggataaccc caagggaagt tttttcaggc
     6181 atcgtgtgta agcagaatat ataagtgctg ttccctggtg cttcctcgct cactcgaccg
     6241 ggagggttcg agaagggggg gcacccccct tcggcgtgcg cggtcacgcg cacagggcgc
     6301 agccctggtt aaaaacaagg tttataaata ttggtttaaa agcaggttaa aagacaggtt
     6361 agcggtggcc gaaaaacggg cggaaaccct tgcaaatgct ggattttctg cctgtggaca
     6421 gcccctcaaa tgtcaatagg tgcgcccctc atctgtcagc actctgcccc tcaagtgtca
     6481 aggatcgcgc ccctcatctg tcagtagtcg cgcccctcaa gtgtcaatac cgcagggcac
     6541 ttatccccag gcttgtccac atcatctgtg ggaaactcgc gtaaaatcag gcgttttcgc
     6601 cgatttgcga ggctggccag ctccacgtcg ccggccgaaa tcgagcctgc ccctcatctg
     6661 tcaacgccgc gccgggtgag tcggcccctc aagtgtcaac gtccgcccct catctgtcag
     6721 tgagggccaa gttttccgcg aggtatccac aacgccggcg gccggccgcg gtgtctcgca
     6781 cacggcttcg acggcgtttc tggcgcgttt gcagggccat agacggccgc cagcccagcg
     6841 gcgagggcaa ccagccgagg gcttcgccct gtcgctcgac tgcggcgagc actactggct
     6901 gtaaaaggac agaccacatc atggttctgt gttcattagg ttgttctgtc cattgctgac
     6961 ataatccgct ccacttcaac gtaacaccgc acgaagattt ctattgttcc tgaaggcata
     7021 ttcaaatcgt tttcgttacc gcttgcaggc atcatgacag aacactactt cctataaacg
     7081 ctacacaggc tcctgagatt aataatgcgg atctctacga taatgggaga ttttcccgac
     7141 tgtttcgttc gcttctcagt ggataacagc cagcttctct gtttaacaga caaaaacagc
     7201 atatccactc agttccacat ttccatataa aggccaaggc atttattctc aggataattg
     7261 tttcagcatc gcaaccgcat cagactccgg catcgcaaac tgcacccggt gccgggcagc
     7321 cacatccagc gcaaaaacct tcgtgtagac ttccgttgaa ctgatggact tatgtcccat
     7381 caggctttgc agaactttca gcggtatacc ggcatacagc atgtgcatcg cataggaatg
     7441 gcggaacgta tgtggtgtga ccggaacaga gaacgtcaca ccgtcagcag cagcggcggc
     7501 aaccgcctcc ccaatccagg tcctgaccgt tctgtccgtc acttcccaga tccgcgcttt
     7561 ctctgtcctt cctgtgcgac ggttacgccg ctccatgagc ttatcgcgaa taaatacctg
     7621 tgacggaaga tcacttcgca gaataaataa atcctggtgt ccctgttgat accgggaagc
     7681 cctgggccaa cttttggcga aaatgagacg ttgatcggca cgtaagaggt tccaactttc
     7741 accataatga aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag
     7801 ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat
     7861 ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga
     7921 ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt
     7981 atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa tttcgtatgg
     8041 caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc
     8101 atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt
     8161 ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta
     8221 aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt
     8281 ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat
     8341 attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt
     8401 gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc
     8461 agggcggggc gtaagacgtc taag
//
