LOCUS       sequence-148757-        9340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Output plasmid: contains 5-state register that regulates expression 
            of GFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9340)
  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 9340)
  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: pNR164
FEATURES             Location/Qualifiers
     source          1..9340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    578..626
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     terminator      932..1003
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    1228..1285
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attP site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     misc_RNA        1329..1378
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             1421..1429
                     /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(1465..1493)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2088..2108
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    2684..3157
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     CDS             complement(3230..4201)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(4201..5376)
                     /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    5702..5952
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(5955..6710)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      6801..7020
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      7088..7619
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      7096..7710
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     promoter        8568..8670
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             8671..9330
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(8737..8756)
                     /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 cgtcgctaac
      301 accgtgcgtt tgactatttt acctctggcg gtgataatgg ttgcatatag tgaacaagga
      361 ttaacgccga gtgcggtatt cctctgggca tgccaacaca attaacatct caatcaaggt
      421 aaatgctttt tgcttttttt gccgtggtgc ggatctagct acccgcactt gagtccacgg
      481 ctgatgccaa cacaattaac atcagaatca aggtaaatgc tttttgcttt ttttgcgggc
      541 tcactaggga atggcgtcgt ccgtgacatt ctgtgcgcgg ccggcttgtc gacgacggcg
      601 gtctccgtcg tcaggatcat ccgggccagc ggctgtctat gccagaagtc ggtcaggtcg
      661 gagtcctaga aaggagtttt ttagttacct taattctaat aaacgaaata aaaactcgcc
      721 gttcgcgcga catgacatgc gccgccgatt gattcagaaa gttaaacaaa attatttgta
      781 gagggaaacc gttgtggtct ccctgaatat attatacgag ccttatgcat gcccgtaaag
      841 ttatccagca accactcata gacctagggc agcagatagg gacgacgtgg tgttagctgt
      901 gacagacctc ccactcacac tccccaggca tcaaataaaa cgaaaggctc agtcgaaaga
      961 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcgggccca ctcgcagctt
     1021 aagttcagcc aaaaaactta agaccgccgg tcttgtccac taccttgcag taatgcggtg
     1081 gacaggatcg gcggttttct tttctcttct caacccgaat gggagtgaac accaacgcat
     1141 gagaaagccc ccggaagatc accttccggg ggctttttta ttgcgcgcat tgagctaccc
     1201 ggtggccgac acaatctccc tgctcaggtc gtggtttgtc tggtcaacca ccgcggtctc
     1261 agtggtgtac ggtacaaacc ccgacgcaat agcgccaacc atgcacgtga ggacgaggga
     1321 tggcacagct gtcaccggat gtgctttccg gtctgatgag tccgtgagga cgaaacagcc
     1381 tctacaaata attttgttta aggtcgaagc cgacctatat taaggaggtc actaatgagt
     1441 aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt
     1501 aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt
     1561 acccttaaat ttatttgcac tactggaaaa ctacctgttc catggccaac acttgtcact
     1621 actttcggtt atggtgttca atgctttgcg agatacccag atcatatgaa acagcatgac
     1681 tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat
     1741 gacgggaact acaagacacg tgctgaagtc aagtttgaag gtgataccct tgttaataga
     1801 atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttcttggaca caaattggaa
     1861 tacaactata actcacacaa tgtatacatc atggcagaca aacaaaagaa tggaatcaaa
     1921 gttaacttca aaattagaca caacattgaa gatggaagcg ttcaactagc agaccattat
     1981 caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc
     2041 acacaatctg ccctttcgaa agatcccaac gaaaagagag accacatggt ccttcttgag
     2101 tttgtaacag ctgctgggat tacacatggc atggatgatc tctacaaata aggaaacaca
     2161 gaaaaaagcc cgcacctgac agtgcgggct ttttttttcg accaaagggt gctttatcaa
     2221 ggggaaggca tagccagcct gacagtagcc aaaggagttt tttagttacc ttaattgaaa
     2281 taaacgaaat aaaaactcgc gcagctgaga gcgacacagt tgcctcactc ctagggtgta
     2341 ataggcacta aaagagtgat atggcacgcg catgtggaac gccgttagag ggttgctgag
     2401 ccatcgtggt ccttagactc gggaaaaaca aaaacccgcc gaagcgggtt aagtgcgggg
     2461 cggctagccc gccacctaag ttatgagtca ggaaaaaagg cgacagagta atctgtcgcc
     2521 ttttttcttt gcttgctttg taatcactcg atgaccaaaa gtaaaaaccc gccgaagcgg
     2581 gtttttacgt aaaacaggtg aaactgacgc tagccggtca catgtcaata cttgcccttg
     2641 acaggcattg atggaatcgt agtctcacgc tgatagtctg atcgacaata caagtgggac
     2701 cgtggtccca gaccgataat cagaccgaca acacgagtgg gatcgtggtc ccagactaat
     2761 aatcagaccg acgatacgag tgggaccgtg gtcccagact aataatcaga ccgacgatac
     2821 gagtgggacc gtggttccag actaataatc agaccgacga tacgagtggg accgtggtcc
     2881 cagactaata atcagaccga cgatacgagt gggaccatgg tcccagacta ataatcagac
     2941 cgacgatacg agtgggaccg tggtcccagt ctgattatca gaccgacgat acgagtggga
     3001 ccgtggtccc agactaataa tcagaccgac gatacgagtg ggaccgtggt cccagactaa
     3061 taatcagacc gacgatacga gtgggaccgt ggtcccagtc tgattatcag accgacgata
     3121 caagtggaac agtgggccca gagagaatat tcaggccagt tatgctttct ggcctgtaac
     3181 aaaggacatt aagtaaagac agataaacgt agactaaaac gtggtcgcat cagggtgctg
     3241 gcttttcaag ttccttaaga atggcctcaa ttttctctat acactcagtt ggaacacggg
     3301 acctgtccag gttaagcacc attttatcgc ccttatacaa tactgtcgct ccaggagcaa
     3361 actgatgtcg tgagcttaaa ctagttcttg atgcagatga cgttttaagc acagaagtta
     3421 aaagagtgat aacttcttca gcttcaaata tcaccccagc ttttttctgc tcatgaaggt
     3481 tagatgcctg ctgcttaagt aattcctctt tatctgtaaa ggctttttga agtgcatcac
     3541 ctgaccgggc agatagttca ccggggtgag aaaaaagagc aacaactgat ttaggcaatt
     3601 tggcggtgtt gatacagcgg gtaataatct tacgtgaaat attttccgca tcagccagcg
     3661 cagaaatatt tccagcaaat tcattctgca atcggcttgc ataacgctga ccacgttcat
     3721 aagcacttgt tgggcgataa tcgttaccca atctggataa tgcagccatc tgctcatcat
     3781 ccagctcgcc aaccagaaca cgataatcac tttcggtaag tgcagcagct ttacgacggc
     3841 gactcccatc ggcaatttct atgacaccag atactcttcg accgaacgcc ggtgtctgtt
     3901 gaccagtcag tagaaaagaa gggatgagat catccagtgc gtcctcagta agcagctcct
     3961 ggtcacgttc attacctgac catacccgag aggtcttctc aacactatca ccccggagca
     4021 cttcaagagt aaacttcaca tcccgaccac atacaggcaa agtaatggca ttaccgcgag
     4081 ccattactcc tacgcgcgca attaacgaat ccaccatcgg ggcagctggt gtcgataacg
     4141 aagtatcttc aaccggttga gtattgagcg tatgttttgg aataacaggc gcacgcttca
     4201 ttatctaatc tcccagcgtg gtttaatcag acgatcgaaa atttcattgc agacaggttc
     4261 ccaaatagaa agagcatttc tccaggcacc agttgaagag cgttgatcaa tggcctgttc
     4321 aaaaacagtt ctcatccgga tctgaccttt accaacttca tccgtttcac gtacaacatt
     4381 ttttagaacc atgcttcccc aggcatcccg aatttgctcc tccatccacg gggactgaga
     4441 gccattacta ttgctgtatt tggtaagcaa aatacgtaca tcaggctcga accctttaag
     4501 atcaacgttc ttgagcagat cacgaagcat atcgaaaaac tgcagtgcgg aggtgtagtc
     4561 aaacaactca gcaggcgtgg gaacaatcag cacatcagca gcacatacga cattaatcgt
     4621 gccgataccc aggttaggcg cgctgtcaat aactatgaca tcatagtcat gagcaacagt
     4681 ttcaatggcc agtcggagca tcaggtgtgg atcggtgggc agtttacctt catcaaattt
     4741 gcccattaac tcagtttcaa tacggtgcag agccagacag gaaggaataa tgtcaagccc
     4801 cggccagcaa gtgggcttta ttgcataagt gacatcgtcc ttttccccaa gatagaaagg
     4861 caggagagtg tcttctgcat gaatatgaag atctggtacc catccgtgat acattgaggc
     4921 tgttccctgg gggtcgttac cttccacgag caaaacacgt agccccttca gagccagatc
     4981 ctgagcaaga tgaacagaaa ctgaggtttt gtaaacgcca cctttatggg cagcaacccc
     5041 gatcaccggt ggaaatacgt cttcagcacg tcgcaatcgc gtaccaaaca catcacgcat
     5101 atgattaatt tgttcaattg tataaccaac acgttgctca acccgtcctc gaatttccat
     5161 atccgggtgc ggtagtcgcc ctgctttctc ggcatctctg atagcctgag aagaaacccc
     5221 aactaaatcc gctgcttcac ctattctcca gcgccgggtt attttcctcg cttccgggct
     5281 gtcatcatta aactgtgcaa tggcgatagc cttcgtcatt tcatgaccag cgtttatgca
     5341 ctggttaagt gtttccatga gtttcattct gaacatcctt taatcattgc tttgcgtttt
     5401 tttattaaat cttgcaattt actgcaaagc aacaacaaaa tcgcaaagtc atcaaaaaac
     5461 cgcaaagttg tttaaaataa gagcaacact acaaaaggag ataagaagag cacatacctc
     5521 agtcacttat tatcactagc gctcgccgca gccgtgtaac cgagcatagc gagcgaactg
     5581 gcgaggaagc aaagaagaac tgttctgtca gatagctctt acgctcagcg caagaagaaa
     5641 tatccaccgt gggaaaaact ccaggtagag gtacacacgc ggatagccaa ttcagagtaa
     5701 taaactgtga taatcaaccc tcatcaatga tgacgaacta acccccgata tcaggtcaca
     5761 tgacgaaggg aaagagaagg aaatcaactg tgacaaactg ccctcaaatt tggcttcctt
     5821 aaaaattaca gttcaaaaag tatgagaaaa tccatgcagg ctgaaggaaa cagcaaaact
     5881 gtgacaaatt accctcagta ggtcagaaca aatgtgacga accaccctca aatctgtgac
     5941 agataaccct cagactatcc tgtcgtcatg gaagtgatat cgcggaagga aaatacgata
     6001 tgagtcgtct ggcggccttt ctttttctca atgtatgaga ggcgcattgg agttctgctg
     6061 ttgatctcat taacacagac ctgcaggaag cggcggcgga agtcaggcat acgctggtaa
     6121 ctttgaggca gctggtaacg ctctatgatc cagtcgattt tcagagagac gatgcctgag
     6181 ccatccggct tacgatactg acacagggat tcgtataaac gcatggcata cggattggtg
     6241 atttcttttg tttcactaag ccgaaactgc gtaaaccggt tctgtaaccc gataaagaag
     6301 ggaatgagat atgggttgat atgtacactg taaagccctc tggatggact gtgcgcacgt
     6361 ttgataaacc aaggaaaaga ttcatagcct ttttcatcgc cggcatcctc ttcagggcga
     6421 taaaaaacca cttccttccc cgcgaaactc ttcaatgcct gccgtatatc cttactggct
     6481 tccgcagagg tcaatccgaa tatttcagca tatttagcaa catggatctc gcagataccg
     6541 tcatgttcct gtagggtgcc atcagatttt ctgatctggt caacgaacag atacagcata
     6601 cgtttttgat cccgggagag actatatgcc gcctcagtga ggtcgtttga ctggacgatt
     6661 cgcgggctat ttttacgttt cttgtgattg ataaccgctg tttccgccat gacagatcca
     6721 tgtgaagtgt gacaagtttt tagattgtca cactaaataa aaaagagtca ataagcaggg
     6781 ataactttgt gaaaaaacag cttcttctga gggcaatttg tcacagggtt aagggcaatt
     6841 tgtcacagac aggactgtca tttgagggtg atttgtcaca ctgaaagggc aatttgtcac
     6901 aacaccttct ctagaaccag catggataaa ggcctacaag gcgctctaaa aaagaagatc
     6961 taaaaactat aaaaaaaata attataaaaa tatccccgtg gataagtgga taaccccaag
     7021 ggaagttttt tcaggcatcg tgtgtaagca gaatatataa gtgctgttcc ctggtgcttc
     7081 ctcgctcact cgaccgggag ggttcgagaa gggggggcac cccccttcgg cgtgcgcggt
     7141 cacgcgcaca gggcgcagcc ctggttaaaa acaaggttta taaatattgg tttaaaagca
     7201 ggttaaaaga caggttagcg gtggccgaaa aacgggcgga aacccttgca aatgctggat
     7261 tttctgcctg tggacagccc ctcaaatgtc aataggtgcg cccctcatct gtcagcactc
     7321 tgcccctcaa gtgtcaagga tcgcgcccct catctgtcag tagtcgcgcc cctcaagtgt
     7381 caataccgca gggcacttat ccccaggctt gtccacatca tctgtgggaa actcgcgtaa
     7441 aatcaggcgt tttcgccgat ttgcgaggct ggccagctcc acgtcgccgg ccgaaatcga
     7501 gcctgcccct catctgtcaa cgccgcgccg ggtgagtcgg cccctcaagt gtcaacgtcc
     7561 gcccctcatc tgtcagtgag ggccaagttt tccgcgaggt atccacaacg ccggcggccg
     7621 gccgcggtgt ctcgcacacg gcttcgacgg cgtttctggc gcgtttgcag ggccatagac
     7681 ggccgccagc ccagcggcga gggcaaccag ccgagggctt cgccctgtcg ctcgactgcg
     7741 gcgagcacta ctggctgtaa aaggacagac cacatcatgg ttctgtgttc attaggttgt
     7801 tctgtccatt gctgacataa tccgctccac ttcaacgtaa caccgcacga agatttctat
     7861 tgttcctgaa ggcatattca aatcgttttc gttaccgctt gcaggcatca tgacagaaca
     7921 ctacttccta taaacgctac acaggctcct gagattaata atgcggatct ctacgataat
     7981 gggagatttt cccgactgtt tcgttcgctt ctcagtggat aacagccagc ttctctgttt
     8041 aacagacaaa aacagcatat ccactcagtt ccacatttcc atataaaggc caaggcattt
     8101 attctcagga taattgtttc agcatcgcaa ccgcatcaga ctccggcatc gcaaactgca
     8161 cccggtgccg ggcagccaca tccagcgcaa aaaccttcgt gtagacttcc gttgaactga
     8221 tggacttatg tcccatcagg ctttgcagaa ctttcagcgg tataccggca tacagcatgt
     8281 gcatcgcata ggaatggcgg aacgtatgtg gtgtgaccgg aacagagaac gtcacaccgt
     8341 cagcagcagc ggcggcaacc gcctccccaa tccaggtcct gaccgttctg tccgtcactt
     8401 cccagatccg cgctttctct gtccttcctg tgcgacggtt acgccgctcc atgagcttat
     8461 cgcgaataaa tacctgtgac ggaagatcac ttcgcagaat aaataaatcc tggtgtccct
     8521 gttgataccg ggaagccctg ggccaacttt tggcgaaaat gagacgttga tcggcacgta
     8581 agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta
     8641 tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc
     8701 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa
     8761 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa
     8821 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat
     8881 ccggaatttc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct
     8941 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac
     9001 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac
     9061 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg
     9121 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
     9181 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
     9241 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
     9301 tactgcgatg agtggcaggg cggggcgtaa gacgtctaag
//
