LOCUS       sequence-251836-        4375 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2-input history dependent program  B then A; 0-0-GFP226.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4375)
  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 4375)
  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: 2SP4
FEATURES             Location/Qualifiers
     source          1..4375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      251..345
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      837..1059
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             1107..2057
                     /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(2404..2423)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2415..3230
                     /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(2488..2507)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     protein_bind    3413..3460
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     protein_bind    3594..3643
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     RBS             3720..3731
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(3738..3740,3741..3743,3744..4375,1..79)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
ORIGIN
        1 cgaaaaaaga gaccatatgg tgctattaga gtttgtgact gcagcaggta ttacacttgg
       61 tatggacgaa ctttataaat aaaacctatc tattcgtttt caacagagta agggtatccg
      121 agttcttaga cgtcaggtgg caaacgcgaa gtaatctttt cggttttaaa gaaaaagggc
      181 agggtggtga caccttgccc gtttttttgc cggagctaga gctgtcagac caagtttacg
      241 agctcgcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
      301 ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag cactagggac
      361 agtaagacgg gtaagcctgt tgatgatacc gctgccttac tgggtgcatt agccagtctg
      421 aatgacctgt cacgggataa tccgaagtgg tcagactgga aaatcagagg gcaggaactg
      481 ctgaacagca aaaagtcaga tagcaccaca tagcagaccc gccataaaac gccctgagaa
      541 gcccgtgacg ggcttttctt gtattatggg tagtttcctt gcatgaatcc ataaaaggcg
      601 cctgtagtgc catttacccc cattcactgc cagagccgtg agcgcagcga actgaatgtc
      661 acgaaaaaga cagcgactca ggtgcctgat ggtcggagac aaaaggaata ttcagcgatt
      721 tgcccgagct tgcgagggtg ctacttaagc ctttagggtt ttaaggtctg ttttgtagag
      781 gagcaaacag cgtttgcgac atccttttgt aatactgcgg aactgactaa agtagtgagt
      841 tatacacagg gctgggatct attcttttta tcttttttta ttctttcttt attctataaa
      901 ttataaccac ttgaatataa acaaaaaaaa cacacaaagg tctagcggaa tttacagagg
      961 gtctagcaga atttacaagt tttccagcaa aggtctagca gaatttacag atacccacaa
     1021 ctcaaaggaa aaggacatgt aattatcatt gactagccca tctcaattgg tatagtgatt
     1081 aaaatcacct agaccaattg agatgtatgt ctgaattagt tgttttcaaa gcaaatgaac
     1141 tagcgattag tcgctatgac ttaacggagc atgaaaccaa gctaatttta tgctgtgtgg
     1201 cactactcaa ccccacgatt gaaaacccta caaggaaaga acggacggta tcgttcactt
     1261 ataaccaata cgctcagatg atgaacatca gtagggaaaa tgcttatggt gtattagcta
     1321 aagcaaccag agagctgatg acgagaactg tggaaatcag gaatcctttg gttaaaggct
     1381 ttgagatttt ccagtggaca aactatgcca agttctcaag cgaaaaatta gaattagttt
     1441 ttagtgaaga gatattgcct tatcttttcc agttaaaaaa attcataaaa tataatctgg
     1501 aacatgttaa gtcttttgaa aacaaatact ctatgaggat ttatgagtgg ttattaaaag
     1561 aactaacaca aaagaaaact cacaaggcaa atatagagat tagccttgat gaatttaagt
     1621 tcatgttaat gcttgaaaat aactaccatg agtttaaaag gcttaaccaa tgggttttga
     1681 aaccaataag taaagattta aacacttaca gcaatatgaa attggtggtt gataagcgag
     1741 gccgcccgac tgatacgttg attttccaag ttgaactaga tagacaaatg gatctcgtaa
     1801 ccgaacttga gaacaaccag ataaaaatga atggtgacaa aataccaaca accattacat
     1861 cagattccta cctacataac ggactaagaa aaacactaca cgatgcttta actgcaaaaa
     1921 ttcagctcac cagttttgag gcaaaatttt tgagtgacat gcaaagtaag tatgatctca
     1981 atggttcgtt ctcatggctc acgcaaaaac aacgaaccac actagagaac atactggcta
     2041 aatacggaag gatctgaggt tcttatggct cttgtatcta tcagtgaagc atcaagacta
     2101 acaaacaaaa gtagaacaac tgttcaccgt tacatatcaa agggaaaact gtccatatgc
     2161 acagatgaaa acggtgtaaa aaagatagat acatcagagc ttttacgagt ttttggtgca
     2221 ttcaaagctg ttcaccatga acagatcgac aatgtaacta ctagagctga tccttcaact
     2281 cagcaaaagt tcgatttatt caacaaagcc acgttgtgtc tcaaaatctc tgatgttaca
     2341 ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta
     2401 atacaagggg tgttatgagc catattcaac gggaaacgtc ttgctcccgt ccgcgcttaa
     2461 actccaacat ggacgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
     2521 caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac
     2581 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtccgtctc aactggctga
     2641 cggagtttat gcctctcccg accatcaagc attttatccg tactcctgat gatgcgtggt
     2701 tactcaccac cgcgattcct gggaaaacag ccttccaggt attagaagaa tatcctgatt
     2761 caggtgaaaa tattgttgat gcgctggccg tgttcctgcg ccggttacat tcgattcctg
     2821 tttgtaattg tccttttaac agcgatcgtg tatttcgtct tgctcaggcg caatcacgca
     2881 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
     2941 aacaagtctg gaaagaaatg cacaagctct tgccattctc accggattca gtcgtcactc
     3001 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
     3061 atgttggacg ggtcggaatc gcagaccgtt accaggacct tgccattctt tggaactgcc
     3121 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     3181 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa taatactagc
     3241 agaaatcatc cttagcgaaa gctaaggatt ttttttatct gattaccgcc tttgagtgag
     3301 ctcttactga agcacgattc tactcggaag tggtcaataa tgcgagtttt tatttcgttt
     3361 atttcaatta aggtaactaa aaaactcctt tcccttacgg cagacctgag tgggtttgta
     3421 ccgtacacca ctgagaccgc ggtggttgac cagacaaacc acgataagaa tagtaataca
     3481 ggatccgaat cgtttcagtt gcctgcaaaa aaagcaaaaa gcatttacct tgattgagat
     3541 gttaattgtg ttggcattcc acaaacatta cgagccggat gattaattgt caagcccgga
     3601 tgatcctgac gacggagacc gccgtcgtcg acaagccggc cgaagggtgt ctcaaggtgc
     3661 gtaccttgac tgatgagtcc gaaaggacga aacacccctc tacaaataat tttgtttaaa
     3721 aagaggagaa atactagatg gtgagcaaag gtgaggaact ttttacagga gtagtaccta
     3781 ttcttgtaga gttagatggt gatgtgaatg gccataaatt tagtgtttct ggtgaaggag
     3841 agggcgatgc gacttatggc aagttgaccc ttaaatttat ttgtactact ggtaaacttc
     3901 ctgttccttg gcctactctc gtgacaactc ttacttacgg agtccaatgc ttctctagat
     3961 acccggacca catgaagcag cacgatttct tcaaaagcgc catgcctgag ggatacgtcc
     4021 aagaaaggac gatcttcttc aaggacgacg ggaactacaa gacacgggcg gaggtgaaat
     4081 ttgaggggga cacgctcgtg aacaggattg agttgaaggg gattgacttc aaggaggacg
     4141 gcaatatact cggacacaaa ttagagtaca actacaactc gcacaacgtc tacattatgg
     4201 ccgataagca gaagaacggc attaaggtta acttcaagat caggcacaat atcgaggatg
     4261 gcagcgttca gcttgcagat cattaccaac agaacacacc gatcggcgac ggcccggtac
     4321 tgttgccgga taaccactac ctgtcgacgc agtctgccct atcgaaagac cctaa
//
