LOCUS       sequence-246253-        5612 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3-input history dependent program 0-0-mKate2-sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5612)
  AUTHORS   Zuniga A, Guiziou S, Mayonove P, Meriem ZB, Camacho M, Moreau V, 
            Ciandrini L, Hersen P, Bonnet J
  TITLE     Rational programming of history-dependent logic in cellular 
            populations.
  JOURNAL   Nat Commun. 2020 Sep 21;11(1):4758. doi: 10.1038/s41467-020-18455-z.
  PUBMED    32958811
REFERENCE   2  (bases 1 to 5612)
  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: "Nat 
            Commun."; date: "2020-09-21"; volume: "11(1):4758. doi"; pages: " 
            10.1038/s41467-020-18455-z"
COMMENT     Sequence Label: 3SP4
FEATURES             Location/Qualifiers
     source          1..5612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    216..263
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     RBS             346..357
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      complement(1163..1190)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    1634..1683
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     RBS             1760..1771
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1778..2491
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGIVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2666..2760
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3252..3474
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             3522..4472
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     primer_bind     complement(4819..4838)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             join(4830..5612,1..33)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4903..4922)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 cagtttcatt tgatgctcga tgagtttttc taataatact agcagaaatc atccttagcg
       61 aaagctaagg atttttttta tctgattacc gcctttgagt gagctcttac tgaagcacga
      121 ttctactcgg aagtggtcaa taatgcgagt ttttatttcg tttatttcaa ttaaggtaac
      181 taaaaaactc ctttccctta cggcagacct gagtgggttt gtaccgtaca ccactgagac
      241 cgcggtggtt gaccagacaa accacgaagt ggtcgtgatc tgaaactcga tcacctgatg
      301 agctcaaggc agagcgaaac cacctctaca aataattttg tttaaaaaga ggagaaatac
      361 tagatgtcag aattaattaa agaaaatatg cacatgaaat tatatatgga aggtactgtc
      421 aacaatcatc atttcaaatg cacatccgaa ggtgaaggta aaccatatga aggcacacaa
      481 acaatgcgca tcaaagcagt tgaaggtgga cccctgccct ttgcgtttga cattctcgca
      541 acgagcttta tgtacgggtc taaaactttt atcaatcaca cccaaggcat tcctgacttt
      601 tttaaacagt cctttcctga aggctttacc tgggaacgtg taacaactta tgaagatggc
      661 ggtgtactta cagcaactca agatacgagt ttacaagatg gctgtctgat ttacaatgtt
      721 aaaatccgtg gcgtaaattt cccgagtaac ggacccgtaa tgcaaaaaaa aactcttggt
      781 tgggaagcat caacagaaac cttatatcct gcggacggtg gcttagaagg acgcgcagac
      841 atggcactga aattagttgg aggcggtcat ttaatctgca acctgaaaac aacctatcgt
      901 tccaaaaaac ccgctaaaaa ccttaaaatg cctggagtat actatgttga tcgtcgctta
      961 gaacgtatta aagaagctga taaagaaacc tacgttgaac aacatgaagt agccgtagcc
     1021 cgttattgtg accttccgtc gaaattagga catcgttgat aacacctggg atttccgaag
     1081 attgactatt ctcagataaa gtaaataata aaaaagccgg attaataatc tggcttttta
     1141 tattctctct ctagtatata aacgcagaaa ggcccacccg aaggtgagcc agtgtgataa
     1201 gaatagtaat acaggatccg aatcgtttca gttgcctccc taatacgcaa gtcgataact
     1261 ctcctgggag cgttgacaac ttgcgcaccc tgatctgttg acaattaatc atccggctcg
     1321 taatgtttgt ggaatgccaa cacaattaac atctcaatca aggtaaatgc tttttgcttt
     1381 ttttgcgggt ccctgagacg caactagcac cgaccgcagc cacagcgtgt agcgctccca
     1441 ggccgtccac tccctgatcc ggcgctcccg gcttatcggt cagtttcacc tgatttacgt
     1501 aaaaacccgc ttcggcgggt ttttgctttt ggaggggcag aaagatgaat gactgtccac
     1561 gacgctatac ccaaaagaaa ccaattattg aaggcctccc taacgggggg cctttttttg
     1621 tttctggtct cccgcccgga tgatcctgac gacggagacc gccgtcgtcg acaagccggc
     1681 cgaagggtgt ctcaaggtgc gtaccttgac tgatgagtcc gaaaggacga aacacccctc
     1741 tacaaataat tttgtttaaa aagaggagaa atactagatg cgtaaaggcg aagagctgtt
     1801 cactggtgtc gtccctattc tggtggaact ggatggtgat gtcaacggtc ataagttttc
     1861 cgtgcgtggc gagggtgaag gtgacgcaac taatggtaaa ctgacgctga agttcatctg
     1921 tactactggt aaactgccgg taccttggcc gactctggta acgacgctga cttatggtgt
     1981 tcagtgcttt gctcgttatc cggaccatat gaagcagcat gacttcttca agtccgccat
     2041 gccggaaggc tatgtgcagg aacgcacgat ttcctttaag gatgacggca cgtacaaaac
     2101 gcgtgcggaa gtgaaatttg aaggcgatac cctggtaaac cgcattgagc tgaaaggcat
     2161 tgactttaaa gaagacggca atatcctggg ccataagctg gaatacaatt ttaacagcca
     2221 caatgtttac atcaccgccg ataaacaaaa aaatggcatt aaagcgaatt ttaaaattcg
     2281 ccacaacgtg gaggatggca tcgtgcagct ggctgatcac taccagcaaa acactccaat
     2341 cggtgatggt cctgttctgc tgccagacaa tcactatctg agcacgcaaa gcgttctgtc
     2401 taaagatccg aacgagaaac gcgatcatat ggttctgctg gagttcgtaa ccgcagcggg
     2461 catcacgcat ggtatggatg aactgtacaa atgataaaac ctatctattc gttttcaaca
     2521 gagtaagggt atccgagttc ttagacgtca ggtggcaaac gcgaagtaat cttttcggtt
     2581 ttaaagaaaa agggcagggt ggtgacacct tgcccgtttt tttgccggag ctagagctgt
     2641 cagaccaagt ttacgagctc gcttggactc ctgttgatag atccagtaat gacctcagaa
     2701 ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat tggtgagaat
     2761 ccaagcacta gggacagtaa gacgggtaag cctgttgatg ataccgctgc cttactgggt
     2821 gcattagcca gtctgaatga cctgtcacgg gataatccga agtggtcaga ctggaaaatc
     2881 agagggcagg aactgctgaa cagcaaaaag tcagatagca ccacatagca gacccgccat
     2941 aaaacgccct gagaagcccg tgacgggctt ttcttgtatt atgggtagtt tccttgcatg
     3001 aatccataaa aggcgcctgt agtgccattt acccccattc actgccagag ccgtgagcgc
     3061 agcgaactga atgtcacgaa aaagacagcg actcaggtgc ctgatggtcg gagacaaaag
     3121 gaatattcag cgatttgccc gagcttgcga gggtgctact taagccttta gggttttaag
     3181 gtctgttttg tagaggagca aacagcgttt gcgacatcct tttgtaatac tgcggaactg
     3241 actaaagtag tgagttatac acagggctgg gatctattct ttttatcttt ttttattctt
     3301 tctttattct ataaattata accacttgaa tataaacaaa aaaaacacac aaaggtctag
     3361 cggaatttac agagggtcta gcagaattta caagttttcc agcaaaggtc tagcagaatt
     3421 tacagatacc cacaactcaa aggaaaagga catgtaatta tcattgacta gcccatctca
     3481 attggtatag tgattaaaat cacctagacc aattgagatg tatgtctgaa ttagttgttt
     3541 tcaaagcaaa tgaactagcg attagtcgct atgacttaac ggagcatgaa accaagctaa
     3601 ttttatgctg tgtggcacta ctcaacccca cgattgaaaa ccctacaagg aaagaacgga
     3661 cggtatcgtt cacttataac caatacgctc agatgatgaa catcagtagg gaaaatgctt
     3721 atggtgtatt agctaaagca accagagagc tgatgacgag aactgtggaa atcaggaatc
     3781 ctttggttaa aggctttgag attttccagt ggacaaacta tgccaagttc tcaagcgaaa
     3841 aattagaatt agtttttagt gaagagatat tgccttatct tttccagtta aaaaaattca
     3901 taaaatataa tctggaacat gttaagtctt ttgaaaacaa atactctatg aggatttatg
     3961 agtggttatt aaaagaacta acacaaaaga aaactcacaa ggcaaatata gagattagcc
     4021 ttgatgaatt taagttcatg ttaatgcttg aaaataacta ccatgagttt aaaaggctta
     4081 accaatgggt tttgaaacca ataagtaaag atttaaacac ttacagcaat atgaaattgg
     4141 tggttgataa gcgaggccgc ccgactgata cgttgatttt ccaagttgaa ctagatagac
     4201 aaatggatct cgtaaccgaa cttgagaaca accagataaa aatgaatggt gacaaaatac
     4261 caacaaccat tacatcagat tcctacctac ataacggact aagaaaaaca ctacacgatg
     4321 ctttaactgc aaaaattcag ctcaccagtt ttgaggcaaa atttttgagt gacatgcaaa
     4381 gtaagtatga tctcaatggt tcgttctcat ggctcacgca aaaacaacga accacactag
     4441 agaacatact ggctaaatac ggaaggatct gaggttctta tggctcttgt atctatcagt
     4501 gaagcatcaa gactaacaaa caaaagtaga acaactgttc accgttacat atcaaaggga
     4561 aaactgtcca tatgcacaga tgaaaacggt gtaaaaaaga tagatacatc agagctttta
     4621 cgagtttttg gtgcattcaa agctgttcac catgaacaga tcgacaatgt aactactaga
     4681 gctgattctt caactcagca aaagttcgat ttattcaaca aagccacgtt gtgtctcaaa
     4741 atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg
     4801 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct
     4861 cccgtccgcg cttaaactcc aacatggacg ctgatttata tgggtataaa tgggctcgcg
     4921 ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag
     4981 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtcc
     5041 gtctcaactg gctgacggag tttatgcctc tcccgaccat caagcatttt atccgtactc
     5101 ctgatgatgc gtggttactc accaccgcga ttcctgggaa aacagccttc caggtattag
     5161 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggccgtgttc ctgcgccggt
     5221 tacattcgat tcctgtttgt aattgtcctt ttaacagcga tcgtgtattt cgtcttgctc
     5281 aggcgcaatc acgcatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     5341 atggctggcc tgttgaacaa gtctggaaag aaatgcacaa gctcttgcca ttctcaccgg
     5401 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     5461 taataggttg tattgatgtt ggacgggtcg gaatcgcaga ccgttaccag gaccttgcca
     5521 ttctttggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     5581 atggtattga taatcctgat atgaataaat tg
//
