LOCUS       sequence-148850-        8186 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A118 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 8186)
  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 8186)
  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: pNR280
FEATURES             Location/Qualifiers
     source          1..8186
                     /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        3902..3951
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             3984..3995
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(4032..4060)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4655..4675
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      4778..4864
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    5224..5697
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     CDS             complement(5770..6741)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(6741..7916)
                     /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 ggccgaactt ttcggatcaa gctatgaagg aagaagaaga aacgagaaac taaatgcaca
     3901 gctgtcaccg gatgtgcttt ccggtctgat gagtccgtga ggacgaaaca gcctctacaa
     3961 ataattttgt ttaagaattc attaaagagg agaaaggtac catgagtaaa ggagaagaac
     4021 ttttcactgg agttgtccca attcttgttg aattagatgg tgatgttaat gggcacaaat
     4081 tttctgtcag tggagagggt gaaggtgatg caacatacgg aaaacttacc cttaaattta
     4141 tttgcactac tggaaaacta cctgttccat ggccaacact tgtcactact ttcggttatg
     4201 gtgttcaatg ctttgcgaga tacccagatc atatgaaaca gcatgacttt ttcaagagtg
     4261 ccatgcccga aggttatgta caggaaagaa ctatattttt caaagatgac gggaactaca
     4321 agacacgtgc tgaagtcaag tttgaaggtg atacccttgt taatagaatc gagttaaaag
     4381 gtattgattt taaagaagat ggaaacattc ttggacacaa attggaatac aactataact
     4441 cacacaatgt atacatcatg gcagacaaac aaaagaatgg aatcaaagtt aacttcaaaa
     4501 ttagacacaa cattgaagat ggaagcgttc aactagcaga ccattatcaa caaaatactc
     4561 caattggcga tggccctgtc cttttaccag acaaccatta cctgtccaca caatctgccc
     4621 tttcgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg
     4681 ctgggattac acatggcatg gatgatctct acaaataaaa gcttgatatc gaattcctgc
     4741 agcccggggg atcccatggt acgcgtgcta gaggcatcaa ataaaacgaa aggctcagtc
     4801 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     4861 aaatccgccg ccctagaatc gagtggcgcc tgcagacgta aaaaaagcgg cgtggttagc
     4921 cgctttttta attgccggac gtacgatcga tgcaagtgtt tagttccctc tttgcgtcca
     4981 acgagagaaa acgaggaact aagctagccc gccacctaag ttatgagtca ggaaaaaagg
     5041 cgacagagta atctgtcgcc ttttttcttt gcttgctttg taatcactcg atgaccaaaa
     5101 gtaaaaaccc gccgaagcgg gtttttacgt aaaacaggtg aaactgacgc tagccggtca
     5161 catgtcaata cttgcccttg acaggcattg atggaatcgt agtctcacgc tgatagtctg
     5221 atcgacaata caagtgggac cgtggtccca gaccgataat cagaccgaca acacgagtgg
     5281 gatcgtggtc ccagactaat aatcagaccg acgatacgag tgggaccgtg gtcccagact
     5341 aataatcaga ccgacgatac gagtgggacc gtggttccag actaataatc agaccgacga
     5401 tacgagtggg accgtggtcc cagactaata atcagaccga cgatacgagt gggaccatgg
     5461 tcccagacta ataatcagac cgacgatacg agtgggaccg tggtcccagt ctgattatca
     5521 gaccgacgat acgagtggga ccgtggtccc agactaataa tcagaccgac gatacgagtg
     5581 ggaccgtggt cccagactaa taatcagacc gacgatacga gtgggaccgt ggtcccagtc
     5641 tgattatcag accgacgata caagtggaac agtgggccca gagagaatat tcaggccagt
     5701 tatgctttct ggcctgtaac aaaggacatt aagtaaagac agataaacgt agactaaaac
     5761 gtggtcgcat cagggtgctg gcttttcaag ttccttaaga atggcctcaa ttttctctat
     5821 acactcagtt ggaacacggg acctgtccag gttaagcacc attttatcgc ccttatacaa
     5881 tactgtcgct ccaggagcaa actgatgtcg tgagcttaaa ctagttcttg atgcagatga
     5941 cgttttaagc acagaagtta aaagagtgat aacttcttca gcttcaaata tcaccccagc
     6001 ttttttctgc tcatgaaggt tagatgcctg ctgcttaagt aattcctctt tatctgtaaa
     6061 ggctttttga agtgcatcac ctgaccgggc agatagttca ccggggtgag aaaaaagagc
     6121 aacaactgat ttaggcaatt tggcggtgtt gatacagcgg gtaataatct tacgtgaaat
     6181 attttccgca tcagccagcg cagaaatatt tccagcaaat tcattctgca atcggcttgc
     6241 ataacgctga ccacgttcat aagcacttgt tgggcgataa tcgttaccca atctggataa
     6301 tgcagccatc tgctcatcat ccagctcgcc aaccagaaca cgataatcac tttcggtaag
     6361 tgcagcagct ttacgacggc gactcccatc ggcaatttct atgacaccag atactcttcg
     6421 accgaacgcc ggtgtctgtt gaccagtcag tagaaaagaa gggatgagat catccagtgc
     6481 gtcctcagta agcagctcct ggtcacgttc attacctgac catacccgag aggtcttctc
     6541 aacactatca ccccggagca cttcaagagt aaacttcaca tcccgaccac atacaggcaa
     6601 agtaatggca ttaccgcgag ccattactcc tacgcgcgca attaacgaat ccaccatcgg
     6661 ggcagctggt gtcgataacg aagtatcttc aaccggttga gtattgagcg tatgttttgg
     6721 aataacaggc gcacgcttca ttatctaatc tcccagcgtg gtttaatcag acgatcgaaa
     6781 atttcattgc agacaggttc ccaaatagaa agagcatttc tccaggcacc agttgaagag
     6841 cgttgatcaa tggcctgttc aaaaacagtt ctcatccgga tctgaccttt accaacttca
     6901 tccgtttcac gtacaacatt ttttagaacc atgcttcccc aggcatcccg aatttgctcc
     6961 tccatccacg gggactgaga gccattacta ttgctgtatt tggtaagcaa aatacgtaca
     7021 tcaggctcga accctttaag atcaacgttc ttgagcagat cacgaagcat atcgaaaaac
     7081 tgcagtgcgg aggtgtagtc aaacaactca gcaggcgtgg gaacaatcag cacatcagca
     7141 gcacatacga cattaatcgt gccgataccc aggttaggcg cgctgtcaat aactatgaca
     7201 tcatagtcat gagcaacagt ttcaatggcc agtcggagca tcaggtgtgg atcggtgggc
     7261 agtttacctt catcaaattt gcccattaac tcagtttcaa tacggtgcag agccagacag
     7321 gaaggaataa tgtcaagccc cggccagcaa gtgggcttta ttgcataagt gacatcgtcc
     7381 ttttccccaa gatagaaagg caggagagtg tcttctgcat gaatatgaag atctggtacc
     7441 catccgtgat acattgaggc tgttccctgg gggtcgttac cttccacgag caaaacacgt
     7501 agccccttca gagccagatc ctgagcaaga tgaacagaaa ctgaggtttt gtaaacgcca
     7561 cctttatggg cagcaacccc gatcaccggt ggaaatacgt cttcagcacg tcgcaatcgc
     7621 gtaccaaaca catcacgcat atgattaatt tgttcaattg tataaccaac acgttgctca
     7681 acccgtcctc gaatttccat atccgggtgc ggtagtcgcc ctgctttctc ggcatctctg
     7741 atagcctgag aagaaacccc aactaaatcc gctgcttcac ctattctcca gcgccgggtt
     7801 attttcctcg cttccgggct gtcatcatta aactgtgcaa tggcgatagc cttcgtcatt
     7861 tcatgaccag cgtttatgca ctggttaagt gtttccatga gtttcattct gaacatcctt
     7921 taatcattgc tttgcgtttt tttattaaat cttgcaattt actgcaaagc aacaacaaaa
     7981 tcgcaaagtc atcaaaaaac cgcaaagttg tttaaaataa gagcaacact acaaaaggag
     8041 ataagaagag cacatacctc agtcacttat tatcactagc gctcgccgca gccgtgtaac
     8101 cgagcatagc gagcgaactg gcgaggaagc aaagaagaac tgttctgtca gatagctctt
     8161 acgctcagcg caagaagaaa tatcca
//
