LOCUS       sequence-242327-        6288 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  2-input history dependent program A then B;BFP-mKate2-sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6288)
  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 6288)
  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: 2SP9
FEATURES             Location/Qualifiers
     source          1..6288
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(47..760)
                     /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
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     RBS             767..778
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    944..993
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     misc_RNA        996..1045
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             1069..1080
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1827..1854
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             2660..2671
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2821..2868
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     terminator      2948..2979
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(3078..3893)
                     /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     3801..3820
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3885..3904
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(4251..5201)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      5249..5471
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5963..6057)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(6213..6242)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 ctcggatacc cttactctgt tgaaaacgaa tagataggtt ttatcatttg tacagttcat
       61 ccataccatg cgtgatgccc gctgcggtta cgaactccag cagaaccata tgatcgcgtt
      121 tctcgttcgg atctttagac agaacgcttt gcgtgctcag atagtgattg tctggcagca
      181 gaacaggacc atcaccgatt ggagtgtttt gctggtagtg atcagccagc tgcacgctgc
      241 catcctccac gttgtggcga attttaaaat tcgctttaat gccatttttt tgtttatcgg
      301 cggtgatgta aacattgtgg ctgttaaaat tgtattccag cttatggccc aggatattgc
      361 cgtcttcttt aaagtcaatg cctttcagct caatgcggtt taccagggta tcgccttcaa
      421 atttcacttc cgcacgcgtt ttgtacgtgc cgtcatcctt aaaggaaatc gtgcgttcct
      481 gcacatagcc ttccggcatg gcggacttga agaagtcatg ctgcttcata tggtccggat
      541 aacgagcaaa gcactgaaca ccataagtca gcgtcgttac cagagtcggc caaggtaccg
      601 gcagtttacc agtagtacag atgaacttca gcgtcagttt accattagtt gcgtcacctt
      661 caccctcgcc acgcacggaa aacttatgac cgttgacatc accatccagt tccaccagaa
      721 tagggacgac accagtgaac agctcttcgc ctttacgcat ctagtatttc tcctcttttt
      781 aaacaaaatt atttgtagag gggtgtttcg tcctttcgga ctcatcagtc aaggtacgca
      841 ccttgagaca ccctatgcca acacaattaa catctcaatc aaggtaaatg ctttttgctt
      901 tttttgcttg acaattaatc atccggctcg taatgtttgt ggatcggccg gcttgtcgac
      961 gacggcggtc tccgtcgtca ggatcatccg ggcagctgtc accggatgtg ctttccggtc
     1021 tgatgagtcc gtgaggacga aacagcctct acaaataatt ttgtttaaaa agaggagaaa
     1081 tactagatga gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc
     1141 gtggacaacc atcacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
     1201 cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
     1261 gctactagct tcctctacgg cagcaagacc ttcatcaacc acacccaggg catccccgac
     1321 ttcttcaagc agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaagac
     1381 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac
     1441 gtcaagatca gaggggtgaa cttcacatcc aacggccctg tgatgcagaa gaaaacactc
     1501 ggctgggagg ccttcaccga gacgctgtac cccgctgacg gcggcctgga aggcagaaac
     1561 gacatggccc tgaagctcgt gggcgggagc catctgatcg caaacatcaa gaccacatat
     1621 agatccaaga aacccgctaa gaacctcaag atgcctggcg tctactatgt ggactacaga
     1681 ctggaaagaa tcaaggaggc caacaacgag acctacgtcg agcagcacga ggtggcagtg
     1741 gccagatact gcgacctccc tagcaaactg gggcactaaa ggcaactgaa acgattcgga
     1801 tcctgtatta ctattcttat cacactggct caccttcggg tgggcctttc tgcgtttata
     1861 tactagagag agaatataaa aagccagatt attaatccgg cttttttatt atttacttta
     1921 tctgagaata gtcaatcttc ggaaatccca ggtgttatca acgatgtcct aatttcgacg
     1981 gaaggtcaca ataacgggct acggctactt catgttgttc aacgtaggtt tctttatcag
     2041 cttctttaat acgttctaag cgacgatcaa catagtatac tccaggcatt ttaaggtttt
     2101 tagcgggttt tttggaacga taggttgttt tcaggttgca gattaaatga ccgcctccaa
     2161 ctaatttcag tgccatgtct gcgcgtcctt ctaagccacc gtccgcagga tataaggttt
     2221 ctgttgatgc ttcccaacca agagtttttt tttgcattac gggtccgtta ctcgggaaat
     2281 ttacgccacg gattttaaca ttgtaaatca gacagccatc ttgtaaactc gtatcttgag
     2341 ttgctgtaag tacaccgcca tcttcataag ttgttacacg ttcccaggta aagccttcag
     2401 gaaaggactg tttaaaaaag tcaggaatgc cttgggtgtg attgataaaa gttttagacc
     2461 cgtacataaa gctcgttgcg agaatgtcaa acgcaaaggg caggggtcca ccttcaactg
     2521 ctttgatgcg cattgtttgt gtgccttcat atggtttacc ttcaccttcg gatgtgcatt
     2581 tgaaatgatg attgttgaca gtaccttcca tatataattt catgtgcata ttttctttaa
     2641 ttaattctga catctagtat ttctcctctt tttaaacaaa attatttgta gaggtggttt
     2701 cgctctgcct tgagctcatc aggtgatcga gtttcagatc acgaccactg cgagttttta
     2761 tttcgtttat ttcaattaag gtaactaaaa aactccttta ctcaggtctg ccgtaagggg
     2821 ggtttgtacc gtacaccact gagaccgcgg tggttgacca gacaaaccac gaattattga
     2881 ccacttccga gtagaatcgt gcttcagtaa gagagtcact aagggttagt tagttagatt
     2941 agcagaaagt caaaagcctc cgaccggagg cttttgacta aaacttccct tggggttatc
     3001 attggggctc actcaaaggc ggtaatcaga taaaaaaaat ccttagcttt cgctaaggat
     3061 gatttctgct agtattatta gaaaaactca tcgagcatca aatgaaactg caatttattc
     3121 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     3181 tcaccgaggc agttccaaag aatggcaagg tcctggtaac ggtctgcgat tccgacccgt
     3241 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     3301 tcaccatgag tgacgactga atccggtgag aatggcaaga gcttgtgcat ttctttccag
     3361 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     3421 ttattcatgc gtgattgcgc ctgagcaaga cgaaatacac gatcgctgtt aaaaggacaa
     3481 ttacaaacag gaatcgaatg taaccggcgc aggaacacgg ccagcgcatc aacaatattt
     3541 tcacctgaat caggatattc ttctaatacc tggaaggctg ttttcccagg aatcgcggtg
     3601 gtgagtaacc acgcatcatc aggagtacgg ataaaatgct tgatggtcgg gagaggcata
     3661 aactccgtca gccagttgag acggaccatc tcatctgtaa catcattggc aacgctacct
     3721 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaagcg atagattgtc
     3781 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcgtccatg
     3841 ttggagttta agcgcggacg ggagcaagac gtttcccgtt gaatatggct cataacaccc
     3901 cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct
     3961 tgtgcaatgt aacatcagag attttgagac acaacgtggc tttgttgaat aaatcgaact
     4021 tttgctgagt tgaaggatca gctctagtag ttacattgtc gatctgttca tggtgaacag
     4081 ctttaaatgc accaaaaact cgtaaaagct ctgatgtatc tatctttttt acaccgtttt
     4141 catctgtgca tatggacagt tttccctttg atatctaacg gtgaacagtt gttctacttt
     4201 tgtttgttag tcttgatgct tcactgatag atacaagagc cataagaacc tcagatcctt
     4261 ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac
     4321 gaaccattga gatcatgctt actttgcatg tcactcaaaa attttgcctc aaaactggtg
     4381 agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt acgtaggtag
     4441 gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg gttgttctca
     4501 agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc
     4561 gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa atctttactt
     4621 attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt ttcaagcatt
     4681 aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt tttcttttgt
     4741 gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc aaaagactta
     4801 acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg caatatctct
     4861 tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt ccactggaaa
     4921 atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat cagctctctg
     4981 gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat ctgagcgtat
     5041 tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat cgtggggttg
     5101 agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc atagcgacta
     5161 atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa ttggtctagg
     5221 tgattttaat cactatacca attgagatgg gctagtcaat gataattaca tgtccttttc
     5281 ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact tgtaaattct
     5341 gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta tattcaagtg
     5401 gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga tcccagccct
     5461 gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc gcaaacgctg
     5521 tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac cctcgcaagc
     5581 tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga gtcgctgtct
     5641 ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg gtaaatggca
     5701 ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag aaaagcccgt
     5761 cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct gactttttgc
     5821 tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta tcccgtgaca
     5881 ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca ggcttacccg
     5941 tcttactgtc cctagtgctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg
     6001 ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa
     6061 gcgagctcgt aaacttggtc tgacagctct agctccggca aaaaaacggg caaggtgtca
     6121 ccaccctgcc ctttttcttt aaaaccgaaa agattacttc gcgtttgcca cctgacgtct
     6181 aagaaaagga atattcagca atttgcccgt gccgaagaaa ggcccacccg tgaaggtgag
     6241 ccagtgagtt gattgctacg taattagtta gttagccctt agtgactc
//
