LOCUS       sequence-148853-        8191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TP901 attL-attR. GFP expression will stop if sites recombine..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8191)
  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 8191)
  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: pNR287
FEATURES             Location/Qualifiers
     source          1..8191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    56..306
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(309..1064)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      1155..1374
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      1442..1973
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      1450..2064
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     promoter        2922..3024
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3025..3684
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3091..3110)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     misc_RNA        3901..3950
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             3983..3994
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(4031..4059)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4654..4674
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      4777..4863
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    5229..5702
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     CDS             complement(5775..6746)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(6746..7921)
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
ORIGIN
        1 ccgtgggaaa aactccaggt agaggtacac acgcggatag ccaattcaga gtaataaact
       61 gtgataatca accctcatca atgatgacga actaaccccc gatatcaggt cacatgacga
      121 agggaaagag aaggaaatca actgtgacaa actgccctca aatttggctt ccttaaaaat
      181 tacagttcaa aaagtatgag aaaatccatg caggctgaag gaaacagcaa aactgtgaca
      241 aattaccctc agtaggtcag aacaaatgtg acgaaccacc ctcaaatctg tgacagataa
      301 ccctcagact atcctgtcgt catggaagtg atatcgcgga aggaaaatac gatatgagtc
      361 gtctggcggc ctttcttttt ctcaatgtat gagaggcgca ttggagttct gctgttgatc
      421 tcattaacac agacctgcag gaagcggcgg cggaagtcag gcatacgctg gtaactttga
      481 ggcagctggt aacgctctat gatccagtcg attttcagag agacgatgcc tgagccatcc
      541 ggcttacgat actgacacag ggattcgtat aaacgcatgg catacggatt ggtgatttct
      601 tttgtttcac taagccgaaa ctgcgtaaac cggttctgta acccgataaa gaagggaatg
      661 agatatgggt tgatatgtac actgtaaagc cctctggatg gactgtgcgc acgtttgata
      721 aaccaaggaa aagattcata gcctttttca tcgccggcat cctcttcagg gcgataaaaa
      781 accacttcct tccccgcgaa actcttcaat gcctgccgta tatccttact ggcttccgca
      841 gaggtcaatc cgaatatttc agcatattta gcaacatgga tctcgcagat accgtcatgt
      901 tcctgtaggg tgccatcaga ttttctgatc tggtcaacga acagatacag catacgtttt
      961 tgatcccggg agagactata tgccgcctca gtgaggtcgt ttgactggac gattcgcggg
     1021 ctatttttac gtttcttgtg attgataacc gctgtttccg ccatgacaga tccatgtgaa
     1081 gtgtgacaag tttttagatt gtcacactaa ataaaaaaga gtcaataagc agggataact
     1141 ttgtgaaaaa acagcttctt ctgagggcaa tttgtcacag ggttaagggc aatttgtcac
     1201 agacaggact gtcatttgag ggtgatttgt cacactgaaa gggcaatttg tcacaacacc
     1261 ttctctagaa ccagcatgga taaaggccta caaggcgctc taaaaaagaa gatctaaaaa
     1321 ctataaaaaa aataattata aaaatatccc cgtggataag tggataaccc caagggaagt
     1381 tttttcaggc atcgtgtgta agcagaatat ataagtgctg ttccctggtg cttcctcgct
     1441 cactcgaccg ggagggttcg agaagggggg gcacccccct tcggcgtgcg cggtcacgcg
     1501 cacagggcgc agccctggtt aaaaacaagg tttataaata ttggtttaaa agcaggttaa
     1561 aagacaggtt agcggtggcc gaaaaacggg cggaaaccct tgcaaatgct ggattttctg
     1621 cctgtggaca gcccctcaaa tgtcaatagg tgcgcccctc atctgtcagc actctgcccc
     1681 tcaagtgtca aggatcgcgc ccctcatctg tcagtagtcg cgcccctcaa gtgtcaatac
     1741 cgcagggcac ttatccccag gcttgtccac atcatctgtg ggaaactcgc gtaaaatcag
     1801 gcgttttcgc cgatttgcga ggctggccag ctccacgtcg ccggccgaaa tcgagcctgc
     1861 ccctcatctg tcaacgccgc gccgggtgag tcggcccctc aagtgtcaac gtccgcccct
     1921 catctgtcag tgagggccaa gttttccgcg aggtatccac aacgccggcg gccggccgcg
     1981 gtgtctcgca cacggcttcg acggcgtttc tggcgcgttt gcagggccat agacggccgc
     2041 cagcccagcg gcgagggcaa ccagccgagg gcttcgccct gtcgctcgac tgcggcgagc
     2101 actactggct gtaaaaggac agaccacatc atggttctgt gttcattagg ttgttctgtc
     2161 cattgctgac ataatccgct ccacttcaac gtaacaccgc acgaagattt ctattgttcc
     2221 tgaaggcata ttcaaatcgt tttcgttacc gcttgcaggc atcatgacag aacactactt
     2281 cctataaacg ctacacaggc tcctgagatt aataatgcgg atctctacga taatgggaga
     2341 ttttcccgac tgtttcgttc gcttctcagt ggataacagc cagcttctct gtttaacaga
     2401 caaaaacagc atatccactc agttccacat ttccatataa aggccaaggc atttattctc
     2461 aggataattg tttcagcatc gcaaccgcat cagactccgg catcgcaaac tgcacccggt
     2521 gccgggcagc cacatccagc gcaaaaacct tcgtgtagac ttccgttgaa ctgatggact
     2581 tatgtcccat caggctttgc agaactttca gcggtatacc ggcatacagc atgtgcatcg
     2641 cataggaatg gcggaacgta tgtggtgtga ccggaacaga gaacgtcaca ccgtcagcag
     2701 cagcggcggc aaccgcctcc ccaatccagg tcctgaccgt tctgtccgtc acttcccaga
     2761 tccgcgcttt ctctgtcctt cctgtgcgac ggttacgccg ctccatgagc ttatcgcgaa
     2821 taaatacctg tgacggaaga tcacttcgca gaataaataa atcctggtgt ccctgttgat
     2881 accgggaagc cctgggccaa cttttggcga aaatgagacg ttgatcggca cgtaagaggt
     2941 tccaactttc accataatga aataagatca ctaccgggcg tattttttga gttatcgaga
     3001 ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat
     3061 atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc
     3121 tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag
     3181 cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa
     3241 tttcgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac
     3301 accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat
     3361 ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc
     3421 tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt
     3481 ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc
     3541 atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat
     3601 catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc
     3661 gatgagtggc agggcggggc gtaagaccac agctaacacc acgtcgtccc tatctgctgc
     3721 cctaggtcta tgagtggttg ctggataact ttacgggcat gcataaggct cgtataatat
     3781 attcagggag accacaacgg tttccctcta caaataattt tgtttaactt tttaattaac
     3841 ggccgatgcc aacacaatta acatctcaat taaggtaact aaaaaactcc ttttgcacag
     3901 ctgtcaccgg atgtgctttc cggtctgatg agtccgtgag gacgaaacag cctctacaaa
     3961 taattttgtt taagaattca ttaaagagga gaaaggtacc atgagtaaag gagaagaact
     4021 tttcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt
     4081 ttctgtcagt ggagagggtg aaggtgatgc aacatacgga aaacttaccc ttaaatttat
     4141 ttgcactact ggaaaactac ctgttccatg gccaacactt gtcactactt tcggttatgg
     4201 tgttcaatgc tttgcgagat acccagatca tatgaaacag catgactttt tcaagagtgc
     4261 catgcccgaa ggttatgtac aggaaagaac tatatttttc aaagatgacg ggaactacaa
     4321 gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg
     4381 tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc
     4441 acacaatgta tacatcatgg cagacaaaca aaagaatgga atcaaagtta acttcaaaat
     4501 tagacacaac attgaagatg gaagcgttca actagcagac cattatcaac aaaatactcc
     4561 aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtccacac aatctgccct
     4621 ttcgaaagat cccaacgaaa agagagacca catggtcctt cttgagtttg taacagctgc
     4681 tgggattaca catggcatgg atgatctcta caaataaaag cttgatatcg aattcctgca
     4741 gcccggggga tcccatggta cgcgtgctag aggcatcaaa taaaacgaaa ggctcagtcg
     4801 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
     4861 aatccgccgc cctagaatcg agtggcgcct gcagacgtaa aaaaagcggc gtggttagcc
     4921 gcttttttaa ttgccggacg tacgatcgat gcgcaaaaaa agcaaaaagc atttaccttg
     4981 attgaaataa acgaaataaa aactcgcgct agcccgccac ctaagttatg agtcaggaaa
     5041 aaaggcgaca gagtaatctg tcgccttttt tctttgcttg ctttgtaatc actcgatgac
     5101 caaaagtaaa aacccgccga agcgggtttt tacgtaaaac aggtgaaact gacgctagcc
     5161 ggtcacatgt caatacttgc ccttgacagg cattgatgga atcgtagtct cacgctgata
     5221 gtctgatcga caatacaagt gggaccgtgg tcccagaccg ataatcagac cgacaacacg
     5281 agtgggatcg tggtcccaga ctaataatca gaccgacgat acgagtggga ccgtggtccc
     5341 agactaataa tcagaccgac gatacgagtg ggaccgtggt tccagactaa taatcagacc
     5401 gacgatacga gtgggaccgt ggtcccagac taataatcag accgacgata cgagtgggac
     5461 catggtccca gactaataat cagaccgacg atacgagtgg gaccgtggtc ccagtctgat
     5521 tatcagaccg acgatacgag tgggaccgtg gtcccagact aataatcaga ccgacgatac
     5581 gagtgggacc gtggtcccag actaataatc agaccgacga tacgagtggg accgtggtcc
     5641 cagtctgatt atcagaccga cgatacaagt ggaacagtgg gcccagagag aatattcagg
     5701 ccagttatgc tttctggcct gtaacaaagg acattaagta aagacagata aacgtagact
     5761 aaaacgtggt cgcatcaggg tgctggcttt tcaagttcct taagaatggc ctcaattttc
     5821 tctatacact cagttggaac acgggacctg tccaggttaa gcaccatttt atcgccctta
     5881 tacaatactg tcgctccagg agcaaactga tgtcgtgagc ttaaactagt tcttgatgca
     5941 gatgacgttt taagcacaga agttaaaaga gtgataactt cttcagcttc aaatatcacc
     6001 ccagcttttt tctgctcatg aaggttagat gcctgctgct taagtaattc ctctttatct
     6061 gtaaaggctt tttgaagtgc atcacctgac cgggcagata gttcaccggg gtgagaaaaa
     6121 agagcaacaa ctgatttagg caatttggcg gtgttgatac agcgggtaat aatcttacgt
     6181 gaaatatttt ccgcatcagc cagcgcagaa atatttccag caaattcatt ctgcaatcgg
     6241 cttgcataac gctgaccacg ttcataagca cttgttgggc gataatcgtt acccaatctg
     6301 gataatgcag ccatctgctc atcatccagc tcgccaacca gaacacgata atcactttcg
     6361 gtaagtgcag cagctttacg acggcgactc ccatcggcaa tttctatgac accagatact
     6421 cttcgaccga acgccggtgt ctgttgacca gtcagtagaa aagaagggat gagatcatcc
     6481 agtgcgtcct cagtaagcag ctcctggtca cgttcattac ctgaccatac ccgagaggtc
     6541 ttctcaacac tatcaccccg gagcacttca agagtaaact tcacatcccg accacataca
     6601 ggcaaagtaa tggcattacc gcgagccatt actcctacgc gcgcaattaa cgaatccacc
     6661 atcggggcag ctggtgtcga taacgaagta tcttcaaccg gttgagtatt gagcgtatgt
     6721 tttggaataa caggcgcacg cttcattatc taatctccca gcgtggttta atcagacgat
     6781 cgaaaatttc attgcagaca ggttcccaaa tagaaagagc atttctccag gcaccagttg
     6841 aagagcgttg atcaatggcc tgttcaaaaa cagttctcat ccggatctga cctttaccaa
     6901 cttcatccgt ttcacgtaca acatttttta gaaccatgct tccccaggca tcccgaattt
     6961 gctcctccat ccacggggac tgagagccat tactattgct gtatttggta agcaaaatac
     7021 gtacatcagg ctcgaaccct ttaagatcaa cgttcttgag cagatcacga agcatatcga
     7081 aaaactgcag tgcggaggtg tagtcaaaca actcagcagg cgtgggaaca atcagcacat
     7141 cagcagcaca tacgacatta atcgtgccga tacccaggtt aggcgcgctg tcaataacta
     7201 tgacatcata gtcatgagca acagtttcaa tggccagtcg gagcatcagg tgtggatcgg
     7261 tgggcagttt accttcatca aatttgccca ttaactcagt ttcaatacgg tgcagagcca
     7321 gacaggaagg aataatgtca agccccggcc agcaagtggg ctttattgca taagtgacat
     7381 cgtccttttc cccaagatag aaaggcagga gagtgtcttc tgcatgaata tgaagatctg
     7441 gtacccatcc gtgatacatt gaggctgttc cctgggggtc gttaccttcc acgagcaaaa
     7501 cacgtagccc cttcagagcc agatcctgag caagatgaac agaaactgag gttttgtaaa
     7561 cgccaccttt atgggcagca accccgatca ccggtggaaa tacgtcttca gcacgtcgca
     7621 atcgcgtacc aaacacatca cgcatatgat taatttgttc aattgtataa ccaacacgtt
     7681 gctcaacccg tcctcgaatt tccatatccg ggtgcggtag tcgccctgct ttctcggcat
     7741 ctctgatagc ctgagaagaa accccaacta aatccgctgc ttcacctatt ctccagcgcc
     7801 gggttatttt cctcgcttcc gggctgtcat cattaaactg tgcaatggcg atagccttcg
     7861 tcatttcatg accagcgttt atgcactggt taagtgtttc catgagtttc attctgaaca
     7921 tcctttaatc attgctttgc gtttttttat taaatcttgc aatttactgc aaagcaacaa
     7981 caaaatcgca aagtcatcaa aaaaccgcaa agttgtttaa aataagagca acactacaaa
     8041 aggagataag aagagcacat acctcagtca cttattatca ctagcgctcg ccgcagccgt
     8101 gtaaccgagc atagcgagcg aactggcgag gaagcaaaga agaactgttc tgtcagatag
     8161 ctcttacgct cagcgcaaga agaaatatcc a
//
