LOCUS       sequence-148758-        8271 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BxbI attB-attP. GFP will express if sites recombine. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8271)
  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 8271)
  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: pNR230
FEATURES             Location/Qualifiers
     source          1..8271
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     45..63
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    236..284
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     terminator      complement(372..458)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(561..581)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     1176..1204
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     RBS             1241..1252
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_RNA        complement(1285..1334)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     protein_bind    1343..1400
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attP site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     misc_feature    1622..2095
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     CDS             complement(2168..3139)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(3139..4314)
                     /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    4640..4890
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(4893..5648)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      5739..5958
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      6026..6557
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      6034..6648
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     promoter        7506..7608
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             7609..8268
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(7675..7694)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tcgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
       61 ggccctttcg tcttcacctc gagcacagct aacaccacgt cgtccctatc tgctgcccta
      121 ggtctatgag tggttgctgg ataactttac gggcatgcat aaggctcgta taatatattc
      181 agggagacca caacggtttc cctctacaaa taattttgtt taacttttta attaacggcc
      241 ggcttgtcga cgacggcggt ctccgtcgtc aggatcatcc gggcatcgat cgtacgtccg
      301 gcaattaaaa aagcggctaa ccacgccgct ttttttacgt ctgcaggcgc cactcgattc
      361 tagggcggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga
      421 aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctctagca cgcgtaccat
      481 gggatccccc gggctgcagg aattcgatat caagctttta tttgtagaga tcatccatgc
      541 catgtgtaat cccagcagct gttacaaact caagaaggac catgtggtct ctcttttcgt
      601 tgggatcttt cgaaagggca gattgtgtgg acaggtaatg gttgtctggt aaaaggacag
      661 ggccatcgcc aattggagta ttttgttgat aatggtctgc tagttgaacg cttccatctt
      721 caatgttgtg tctaattttg aagttaactt tgattccatt cttttgtttg tctgccatga
      781 tgtatacatt gtgtgagtta tagttgtatt ccaatttgtg tccaagaatg tttccatctt
      841 ctttaaaatc aatacctttt aactcgattc tattaacaag ggtatcacct tcaaacttga
      901 cttcagcacg tgtcttgtag ttcccgtcat ctttgaaaaa tatagttctt tcctgtacat
      961 aaccttcggg catggcactc ttgaaaaagt catgctgttt catatgatct gggtatctcg
     1021 caaagcattg aacaccataa ccgaaagtag tgacaagtgt tggccatgga acaggtagtt
     1081 ttccagtagt gcaaataaat ttaagggtaa gttttccgta tgttgcatca ccttcaccct
     1141 ctccactgac agaaaatttg tgcccattaa catcaccatc taattcaaca agaattggga
     1201 caactccagt gaaaagttct tctcctttac tcatggtacc tttctcctct ttaatgaatt
     1261 cttaaacaaa attatttgta gaggctgttt cgtcctcacg gactcatcag accggaaagc
     1321 acatccggtg acagctgtgc acgtcggggt ttgtaccgta caccactgag accgcggtgg
     1381 ttgaccagac aaaccacgac gctagcccgc cacctaagtt atgagtcagg aaaaaaggcg
     1441 acagagtaat ctgtcgcctt ttttctttgc ttgctttgta atcactcgat gaccaaaagt
     1501 aaaaacccgc cgaagcgggt ttttacgtaa aacaggtgaa actgacgcta gccggtcaca
     1561 tgtcaatact tgcccttgac aggcattgat ggaatcgtag tctcacgctg atagtctgat
     1621 cgacaataca agtgggaccg tggtcccaga ccgataatca gaccgacaac acgagtggga
     1681 tcgtggtccc agactaataa tcagaccgac gatacgagtg ggaccgtggt cccagactaa
     1741 taatcagacc gacgatacga gtgggaccgt ggttccagac taataatcag accgacgata
     1801 cgagtgggac cgtggtccca gactaataat cagaccgacg atacgagtgg gaccatggtc
     1861 ccagactaat aatcagaccg acgatacgag tgggaccgtg gtcccagtct gattatcaga
     1921 ccgacgatac gagtgggacc gtggtcccag actaataatc agaccgacga tacgagtggg
     1981 accgtggtcc cagactaata atcagaccga cgatacgagt gggaccgtgg tcccagtctg
     2041 attatcagac cgacgataca agtggaacag tgggcccaga gagaatattc aggccagtta
     2101 tgctttctgg cctgtaacaa aggacattaa gtaaagacag ataaacgtag actaaaacgt
     2161 ggtcgcatca gggtgctggc ttttcaagtt ccttaagaat ggcctcaatt ttctctatac
     2221 actcagttgg aacacgggac ctgtccaggt taagcaccat tttatcgccc ttatacaata
     2281 ctgtcgctcc aggagcaaac tgatgtcgtg agcttaaact agttcttgat gcagatgacg
     2341 ttttaagcac agaagttaaa agagtgataa cttcttcagc ttcaaatatc accccagctt
     2401 ttttctgctc atgaaggtta gatgcctgct gcttaagtaa ttcctcttta tctgtaaagg
     2461 ctttttgaag tgcatcacct gaccgggcag atagttcacc ggggtgagaa aaaagagcaa
     2521 caactgattt aggcaatttg gcggtgttga tacagcgggt aataatctta cgtgaaatat
     2581 tttccgcatc agccagcgca gaaatatttc cagcaaattc attctgcaat cggcttgcat
     2641 aacgctgacc acgttcataa gcacttgttg ggcgataatc gttacccaat ctggataatg
     2701 cagccatctg ctcatcatcc agctcgccaa ccagaacacg ataatcactt tcggtaagtg
     2761 cagcagcttt acgacggcga ctcccatcgg caatttctat gacaccagat actcttcgac
     2821 cgaacgccgg tgtctgttga ccagtcagta gaaaagaagg gatgagatca tccagtgcgt
     2881 cctcagtaag cagctcctgg tcacgttcat tacctgacca tacccgagag gtcttctcaa
     2941 cactatcacc ccggagcact tcaagagtaa acttcacatc ccgaccacat acaggcaaag
     3001 taatggcatt accgcgagcc attactccta cgcgcgcaat taacgaatcc accatcgggg
     3061 cagctggtgt cgataacgaa gtatcttcaa ccggttgagt attgagcgta tgttttggaa
     3121 taacaggcgc acgcttcatt atctaatctc ccagcgtggt ttaatcagac gatcgaaaat
     3181 ttcattgcag acaggttccc aaatagaaag agcatttctc caggcaccag ttgaagagcg
     3241 ttgatcaatg gcctgttcaa aaacagttct catccggatc tgacctttac caacttcatc
     3301 cgtttcacgt acaacatttt ttagaaccat gcttccccag gcatcccgaa tttgctcctc
     3361 catccacggg gactgagagc cattactatt gctgtatttg gtaagcaaaa tacgtacatc
     3421 aggctcgaac cctttaagat caacgttctt gagcagatca cgaagcatat cgaaaaactg
     3481 cagtgcggag gtgtagtcaa acaactcagc aggcgtggga acaatcagca catcagcagc
     3541 acatacgaca ttaatcgtgc cgatacccag gttaggcgcg ctgtcaataa ctatgacatc
     3601 atagtcatga gcaacagttt caatggccag tcggagcatc aggtgtggat cggtgggcag
     3661 tttaccttca tcaaatttgc ccattaactc agtttcaata cggtgcagag ccagacagga
     3721 aggaataatg tcaagccccg gccagcaagt gggctttatt gcataagtga catcgtcctt
     3781 ttccccaaga tagaaaggca ggagagtgtc ttctgcatga atatgaagat ctggtaccca
     3841 tccgtgatac attgaggctg ttccctgggg gtcgttacct tccacgagca aaacacgtag
     3901 ccccttcaga gccagatcct gagcaagatg aacagaaact gaggttttgt aaacgccacc
     3961 tttatgggca gcaaccccga tcaccggtgg aaatacgtct tcagcacgtc gcaatcgcgt
     4021 accaaacaca tcacgcatat gattaatttg ttcaattgta taaccaacac gttgctcaac
     4081 ccgtcctcga atttccatat ccgggtgcgg tagtcgccct gctttctcgg catctctgat
     4141 agcctgagaa gaaaccccaa ctaaatccgc tgcttcacct attctccagc gccgggttat
     4201 tttcctcgct tccgggctgt catcattaaa ctgtgcaatg gcgatagcct tcgtcatttc
     4261 atgaccagcg tttatgcact ggttaagtgt ttccatgagt ttcattctga acatccttta
     4321 atcattgctt tgcgtttttt tattaaatct tgcaatttac tgcaaagcaa caacaaaatc
     4381 gcaaagtcat caaaaaaccg caaagttgtt taaaataaga gcaacactac aaaaggagat
     4441 aagaagagca catacctcag tcacttatta tcactagcgc tcgccgcagc cgtgtaaccg
     4501 agcatagcga gcgaactggc gaggaagcaa agaagaactg ttctgtcaga tagctcttac
     4561 gctcagcgca agaagaaata tccaccgtgg gaaaaactcc aggtagaggt acacacgcgg
     4621 atagccaatt cagagtaata aactgtgata atcaaccctc atcaatgatg acgaactaac
     4681 ccccgatatc aggtcacatg acgaagggaa agagaaggaa atcaactgtg acaaactgcc
     4741 ctcaaatttg gcttccttaa aaattacagt tcaaaaagta tgagaaaatc catgcaggct
     4801 gaaggaaaca gcaaaactgt gacaaattac cctcagtagg tcagaacaaa tgtgacgaac
     4861 caccctcaaa tctgtgacag ataaccctca gactatcctg tcgtcatgga agtgatatcg
     4921 cggaaggaaa atacgatatg agtcgtctgg cggcctttct ttttctcaat gtatgagagg
     4981 cgcattggag ttctgctgtt gatctcatta acacagacct gcaggaagcg gcggcggaag
     5041 tcaggcatac gctggtaact ttgaggcagc tggtaacgct ctatgatcca gtcgattttc
     5101 agagagacga tgcctgagcc atccggctta cgatactgac acagggattc gtataaacgc
     5161 atggcatacg gattggtgat ttcttttgtt tcactaagcc gaaactgcgt aaaccggttc
     5221 tgtaacccga taaagaaggg aatgagatat gggttgatat gtacactgta aagccctctg
     5281 gatggactgt gcgcacgttt gataaaccaa ggaaaagatt catagccttt ttcatcgccg
     5341 gcatcctctt cagggcgata aaaaaccact tccttccccg cgaaactctt caatgcctgc
     5401 cgtatatcct tactggcttc cgcagaggtc aatccgaata tttcagcata tttagcaaca
     5461 tggatctcgc agataccgtc atgttcctgt agggtgccat cagattttct gatctggtca
     5521 acgaacagat acagcatacg tttttgatcc cgggagagac tatatgccgc ctcagtgagg
     5581 tcgtttgact ggacgattcg cgggctattt ttacgtttct tgtgattgat aaccgctgtt
     5641 tccgccatga cagatccatg tgaagtgtga caagttttta gattgtcaca ctaaataaaa
     5701 aagagtcaat aagcagggat aactttgtga aaaaacagct tcttctgagg gcaatttgtc
     5761 acagggttaa gggcaatttg tcacagacag gactgtcatt tgagggtgat ttgtcacact
     5821 gaaagggcaa tttgtcacaa caccttctct agaaccagca tggataaagg cctacaaggc
     5881 gctctaaaaa agaagatcta aaaactataa aaaaaataat tataaaaata tccccgtgga
     5941 taagtggata accccaaggg aagttttttc aggcatcgtg tgtaagcaga atatataagt
     6001 gctgttccct ggtgcttcct cgctcactcg accgggaggg ttcgagaagg gggggcaccc
     6061 cccttcggcg tgcgcggtca cgcgcacagg gcgcagccct ggttaaaaac aaggtttata
     6121 aatattggtt taaaagcagg ttaaaagaca ggttagcggt ggccgaaaaa cgggcggaaa
     6181 cccttgcaaa tgctggattt tctgcctgtg gacagcccct caaatgtcaa taggtgcgcc
     6241 cctcatctgt cagcactctg cccctcaagt gtcaaggatc gcgcccctca tctgtcagta
     6301 gtcgcgcccc tcaagtgtca ataccgcagg gcacttatcc ccaggcttgt ccacatcatc
     6361 tgtgggaaac tcgcgtaaaa tcaggcgttt tcgccgattt gcgaggctgg ccagctccac
     6421 gtcgccggcc gaaatcgagc ctgcccctca tctgtcaacg ccgcgccggg tgagtcggcc
     6481 cctcaagtgt caacgtccgc ccctcatctg tcagtgaggg ccaagttttc cgcgaggtat
     6541 ccacaacgcc ggcggccggc cgcggtgtct cgcacacggc ttcgacggcg tttctggcgc
     6601 gtttgcaggg ccatagacgg ccgccagccc agcggcgagg gcaaccagcc gagggcttcg
     6661 ccctgtcgct cgactgcggc gagcactact ggctgtaaaa ggacagacca catcatggtt
     6721 ctgtgttcat taggttgttc tgtccattgc tgacataatc cgctccactt caacgtaaca
     6781 ccgcacgaag atttctattg ttcctgaagg catattcaaa tcgttttcgt taccgcttgc
     6841 aggcatcatg acagaacact acttcctata aacgctacac aggctcctga gattaataat
     6901 gcggatctct acgataatgg gagattttcc cgactgtttc gttcgcttct cagtggataa
     6961 cagccagctt ctctgtttaa cagacaaaaa cagcatatcc actcagttcc acatttccat
     7021 ataaaggcca aggcatttat tctcaggata attgtttcag catcgcaacc gcatcagact
     7081 ccggcatcgc aaactgcacc cggtgccggg cagccacatc cagcgcaaaa accttcgtgt
     7141 agacttccgt tgaactgatg gacttatgtc ccatcaggct ttgcagaact ttcagcggta
     7201 taccggcata cagcatgtgc atcgcatagg aatggcggaa cgtatgtggt gtgaccggaa
     7261 cagagaacgt cacaccgtca gcagcagcgg cggcaaccgc ctccccaatc caggtcctga
     7321 ccgttctgtc cgtcacttcc cagatccgcg ctttctctgt ccttcctgtg cgacggttac
     7381 gccgctccat gagcttatcg cgaataaata cctgtgacgg aagatcactt cgcagaataa
     7441 ataaatcctg gtgtccctgt tgataccggg aagccctggg ccaacttttg gcgaaaatga
     7501 gacgttgatc ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg
     7561 ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa
     7621 atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca
     7681 tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt
     7741 ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc
     7801 cgcctgatga atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata
     7861 tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg
     7921 ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg
     7981 gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc
     8041 gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac
     8101 aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg
     8161 atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg
     8221 cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaaga c
//
