LOCUS       sequence-250102-        4375 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2-input history dependent program A then B; 0-0-sfGFP.
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: 2SP6
FEATURES             Location/Qualifiers
     source          1..4375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    21..70
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     RBS             236..247
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             254..967
                     /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"
     terminator      1142..1236
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1728..1950
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             1998..2948
                     /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(3295..3314)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3306..4121
                     /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(3379..3398)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     protein_bind    4237..4284
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
ORIGIN
        1 ccgaatcgtt tcagttgcct gcccggatga tcctgacgac ggagaccgcc gtcgtcgaca
       61 agccggccga tccacaaaca ttacgagccg gatgattaat tgtcaagcaa aaaaagcaaa
      121 aagcatttac cttgattgag atgttaattg tgttggcata gggtgtctca aggtgcgtac
      181 cttgactgat gagtccgaaa ggacgaaaca cccctctaca aataattttg tttaaaaaga
      241 ggagaaatac tagatgcgta aaggcgaaga gctgttcact ggtgtcgtcc ctattctggt
      301 ggaactggat ggtgatgtca acggtcataa gttttccgtg cgtggcgagg gtgaaggtga
      361 cgcaactaat ggtaaactga cgctgaagtt catctgtact actggtaaac tgccggtacc
      421 ttggccgact ctggtaacga cgctgactta tggtgttcag tgctttgctc gttatccgga
      481 ccatatgaag cagcatgact tcttcaagtc cgccatgccg gaaggctatg tgcaggaacg
      541 cacgatttcc tttaaggatg acggcacgta caaaacgcgt gcggaagtga aatttgaagg
      601 cgataccctg gtaaaccgca ttgagctgaa aggcattgac tttaaagaag acggcaatat
      661 cctgggccat aagctggaat acaattttaa cagccacaat gtttacatca ccgccgataa
      721 acaaaaaaat ggcattaaag cgaattttaa aattcgccac aacgtggagg atggcagcgt
      781 gcagctggct gatcactacc agcaaaacac tccaatcggt gatggtcctg ttctgctgcc
      841 agacaatcac tatctgagca cgcaaagcgt tctgtctaaa gatccgaacg agaaacgcga
      901 tcatatggtt ctgctggagt tcgtaaccgc agcgggcatc acgcatggta tggatgaact
      961 gtacaaatga taaaacctat ctattcgttt tcaacagagt aagggtatcc gagttcttag
     1021 acgtcaggtg gcaaacgcga agtaatcttt tcggttttaa agaaaaaggg cagggtggtg
     1081 acaccttgcc cgtttttttg ccggagctag agctgtcaga ccaagtttac gagctcgctt
     1141 ggactcctgt tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg
     1201 ctcggttgcc gccgggcgtt ttttattggt gagaatccaa gcactaggga cagtaagacg
     1261 ggtaagcctg ttgatgatac cgctgcctta ctgggtgcat tagccagtct gaatgacctg
     1321 tcacgggata atccgaagtg gtcagactgg aaaatcagag ggcaggaact gctgaacagc
     1381 aaaaagtcag atagcaccac atagcagacc cgccataaaa cgccctgaga agcccgtgac
     1441 gggcttttct tgtattatgg gtagtttcct tgcatgaatc cataaaaggc gcctgtagtg
     1501 ccatttaccc ccattcactg ccagagccgt gagcgcagcg aactgaatgt cacgaaaaag
     1561 acagcgactc aggtgcctga tggtcggaga caaaaggaat attcagcgat ttgcccgagc
     1621 ttgcgagggt gctacttaag cctttagggt tttaaggtct gttttgtaga ggagcaaaca
     1681 gcgtttgcga catccttttg taatactgcg gaactgacta aagtagtgag ttatacacag
     1741 ggctgggatc tattcttttt atcttttttt attctttctt tattctataa attataacca
     1801 cttgaatata aacaaaaaaa acacacaaag gtctagcgga atttacagag ggtctagcag
     1861 aatttacaag ttttccagca aaggtctagc agaatttaca gatacccaca actcaaagga
     1921 aaaggacatg taattatcat tgactagccc atctcaattg gtatagtgat taaaatcacc
     1981 tagaccaatt gagatgtatg tctgaattag ttgttttcaa agcaaatgaa ctagcgatta
     2041 gtcgctatga cttaacggag catgaaacca agctaatttt atgctgtgtg gcactactca
     2101 accccacgat tgaaaaccct acaaggaaag aacggacggt atcgttcact tataaccaat
     2161 acgctcagat gatgaacatc agtagggaaa atgcttatgg tgtattagct aaagcaacca
     2221 gagagctgat gacgagaact gtggaaatca ggaatccttt ggttaaaggc tttgagattt
     2281 tccagtggac aaactatgcc aagttctcaa gcgaaaaatt agaattagtt tttagtgaag
     2341 agatattgcc ttatcttttc cagttaaaaa aattcataaa atataatctg gaacatgtta
     2401 agtcttttga aaacaaatac tctatgagga tttatgagtg gttattaaaa gaactaacac
     2461 aaaagaaaac tcacaaggca aatatagaga ttagccttga tgaatttaag ttcatgttaa
     2521 tgcttgaaaa taactaccat gagtttaaaa ggcttaacca atgggttttg aaaccaataa
     2581 gtaaagattt aaacacttac agcaatatga aattggtggt tgataagcga ggccgcccga
     2641 ctgatacgtt gattttccaa gttgaactag atagacaaat ggatctcgta accgaacttg
     2701 agaacaacca gataaaaatg aatggtgaca aaataccaac aaccattaca tcagattcct
     2761 acctacataa cggactaaga aaaacactac acgatgcttt aactgcaaaa attcagctca
     2821 ccagttttga ggcaaaattt ttgagtgaca tgcaaagtaa gtatgatctc aatggttcgt
     2881 tctcatggct cacgcaaaaa caacgaacca cactagagaa catactggct aaatacggaa
     2941 ggatctgagg ttcttatggc tcttgtatct atcagtgaag catcaagact aacaaacaaa
     3001 agtagaacaa ctgttcaccg ttacatatca aagggaaaac tgtccatatg cacagatgaa
     3061 aacggtgtaa aaaagataga tacatcagag cttttacgag tttttggtgc attcaaagct
     3121 gttcaccatg aacagatcga caatgtaact actagagctg atccttcaac tcagcaaaag
     3181 ttcgatttat tcaacaaagc cacgttgtgt ctcaaaatct ctgatgttac attgcacaag
     3241 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     3301 gtgttatgag ccatattcaa cgggaaacgt cttgctcccg tccgcgctta aactccaaca
     3361 tggacgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     3421 caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     3481 gtagcgttgc caatgatgtt acagatgaga tggtccgtct caactggctg acggagttta
     3541 tgcctctccc gaccatcaag cattttatcc gtactcctga tgatgcgtgg ttactcacca
     3601 ccgcgattcc tgggaaaaca gccttccagg tattagaaga atatcctgat tcaggtgaaa
     3661 atattgttga tgcgctggcc gtgttcctgc gccggttaca ttcgattcct gtttgtaatt
     3721 gtccttttaa cagcgatcgt gtatttcgtc ttgctcaggc gcaatcacgc atgaataacg
     3781 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     3841 ggaaagaaat gcacaagctc ttgccattct caccggattc agtcgtcact catggtgatt
     3901 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     3961 gggtcggaat cgcagaccgt taccaggacc ttgccattct ttggaactgc ctcggtgagt
     4021 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     4081 ataaattgca gtttcatttg atgctcgatg agtttttcta ataatactag cagaaatcat
     4141 ccttagcgaa agctaaggat tttttttatc tgattaccgc ctttgagtga gctcttactg
     4201 aagcacgatt ctactcggaa gtggtcaata attcgtggtt tgtctggtca accaccgcgg
     4261 tctcagtggt gtacggtaca aacccccctt acggcagacc tgagtaaagg agttttttag
     4321 ttaccttaat tgaaataaac gaaataaaaa ctcgctaaga atagtaatac aggat
//
