LOCUS       sequence-249682-        5296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2-input history dependent program A then B;BFP-0-sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5296)
  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 5296)
  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: 2SP8
FEATURES             Location/Qualifiers
     source          1..5296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(103..130)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             877..888
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_RNA        complement(912..961)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     protein_bind    964..1013
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     RBS             1179..1190
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1197..1910
                     /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      2085..2179
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2671..2893
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2941..3891
                     /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(4238..4257)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4249..5064
                     /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(4322..4341)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     protein_bind    5180..5227
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
ORIGIN
        1 gccacctggg atttccgaag attgactatt ctcagataaa gtaaataata aaaaagccgg
       61 attaataatc tggcttttta tattctctct ctagtatata aacgcagaaa ggcccacccg
      121 aaggtgagcc agtgtgataa gaatagtaat acaggatccg aatcgtttca gttgccttta
      181 gtgccccagt ttgctaggga ggtcgcagta tctggccact gccacctcgt gctgctcgac
      241 gtaggtctcg ttgttggcct ccttgattct ttccagtctg tagtccacat agtagacgcc
      301 aggcatcttg aggttcttag cgggtttctt ggatctatat gtggtcttga tgtttgcgat
      361 cagatggctc ccgcccacga gcttcagggc catgtcgttt ctgccttcca ggccgccgtc
      421 agcggggtac agcgtctcgg tgaaggcctc ccagccgagt gttttcttct gcatcacagg
      481 gccgttggat gtgaagttca cccctctgat cttgacgttg tagatgaggc agccgtcctg
      541 gaggctggtg tcctgggtag cggtcagcac gcccccgtct tcgtatgtgg tgactctctc
      601 ccatgtgaag ccctcaggga aggactgctt gaagaagtcg gggatgccct gggtgtggtt
      661 gatgaaggtc ttgctgccgt agaggaagct agtagccagg atgtcgaagg cgaaggggag
      721 agggccgccc tcgaccacct tgattctcat ggtctgggtg ccctcgtagg gcttgccttc
      781 gccctcggat gtgcacttga agtgatggtt gtccacggtg ccctccatgt acagcttcat
      841 gtgcatgttc tccttaatca gctcgctcat ctagtatttc tcctcttttt aaacaaaatt
      901 atttgtagag gctgtttcgt cctcacggac tcatcagacc ggaaagcaca tccggtgaca
      961 gctgcccgga tgatcctgac gacggagacc gccgtcgtcg acaagccggc cgatccacaa
     1021 acattacgag ccggatgatt aattgtcaag caaaaaaagc aaaaagcatt taccttgatt
     1081 gagatgttaa ttgtgttggc atagggtgtc tcaaggtgcg taccttgact gatgagtccg
     1141 aaaggacgaa acacccctct acaaataatt ttgtttaaaa agaggagaaa tactagatgc
     1201 gtaaaggcga agagctgttc actggtgtcg tccctattct ggtggaactg gatggtgatg
     1261 tcaacggtca taagttttcc gtgcgtggcg agggtgaagg tgacgcaact aatggtaaac
     1321 tgacgctgaa gttcatctgt actactggta aactgccggt accttggccg actctggtaa
     1381 cgacgctgac ttatggtgtt cagtgctttg ctcgttatcc ggaccatatg aagcagcatg
     1441 acttcttcaa gtccgccatg ccggaaggct atgtgcagga acgcacgatt tcctttaagg
     1501 atgacggcac gtacaaaacg cgtgcggaag tgaaatttga aggcgatacc ctggtaaacc
     1561 gcattgagct gaaaggcatt gactttaaag aagacggcaa tatcctgggc cataagctgg
     1621 aatacaattt taacagccac aatgtttaca tcaccgccga taaacaaaaa aatggcatta
     1681 aagcgaattt taaaattcgc cacaacgtgg aggatggcag cgtgcagctg gctgatcact
     1741 accagcaaaa cactccaatc ggtgatggtc ctgttctgct gccagacaat cactatctga
     1801 gcacgcaaag cgttctgtct aaagatccga acgagaaacg cgatcatatg gttctgctgg
     1861 agttcgtaac cgcagcgggc atcacgcatg gtatggatga actgtacaaa tgataaaacc
     1921 tatctattcg ttttcaacag agtaagggta tccgagttct tagacgtcag gtggcaaacg
     1981 cgaagtaatc ttttcggttt taaagaaaaa gggcagggtg gtgacacctt gcccgttttt
     2041 ttgccggagc tagagctgtc agaccaagtt tacgagctcg cttggactcc tgttgataga
     2101 tccagtaatg acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc
     2161 gttttttatt ggtgagaatc caagcactag ggacagtaag acgggtaagc ctgttgatga
     2221 taccgctgcc ttactgggtg cattagccag tctgaatgac ctgtcacggg ataatccgaa
     2281 gtggtcagac tggaaaatca gagggcagga actgctgaac agcaaaaagt cagatagcac
     2341 cacatagcag acccgccata aaacgccctg agaagcccgt gacgggcttt tcttgtatta
     2401 tgggtagttt ccttgcatga atccataaaa ggcgcctgta gtgccattta cccccattca
     2461 ctgccagagc cgtgagcgca gcgaactgaa tgtcacgaaa aagacagcga ctcaggtgcc
     2521 tgatggtcgg agacaaaagg aatattcagc gatttgcccg agcttgcgag ggtgctactt
     2581 aagcctttag ggttttaagg tctgttttgt agaggagcaa acagcgtttg cgacatcctt
     2641 ttgtaatact gcggaactga ctaaagtagt gagttataca cagggctggg atctattctt
     2701 tttatctttt tttattcttt ctttattcta taaattataa ccacttgaat ataaacaaaa
     2761 aaaacacaca aaggtctagc ggaatttaca gagggtctag cagaatttac aagttttcca
     2821 gcaaaggtct agcagaattt acagataccc acaactcaaa ggaaaaggac atgtaattat
     2881 cattgactag cccatctcaa ttggtatagt gattaaaatc acctagacca attgagatgt
     2941 atgtctgaat tagttgtttt caaagcaaat gaactagcga ttagtcgcta tgacttaacg
     3001 gagcatgaaa ccaagctaat tttatgctgt gtggcactac tcaaccccac gattgaaaac
     3061 cctacaagga aagaacggac ggtatcgttc acttataacc aatacgctca gatgatgaac
     3121 atcagtaggg aaaatgctta tggtgtatta gctaaagcaa ccagagagct gatgacgaga
     3181 actgtggaaa tcaggaatcc tttggttaaa ggctttgaga ttttccagtg gacaaactat
     3241 gccaagttct caagcgaaaa attagaatta gtttttagtg aagagatatt gccttatctt
     3301 ttccagttaa aaaaattcat aaaatataat ctggaacatg ttaagtcttt tgaaaacaaa
     3361 tactctatga ggatttatga gtggttatta aaagaactaa cacaaaagaa aactcacaag
     3421 gcaaatatag agattagcct tgatgaattt aagttcatgt taatgcttga aaataactac
     3481 catgagttta aaaggcttaa ccaatgggtt ttgaaaccaa taagtaaaga tttaaacact
     3541 tacagcaata tgaaattggt ggttgataag cgaggccgcc cgactgatac gttgattttc
     3601 caagttgaac tagatagaca aatggatctc gtaaccgaac ttgagaacaa ccagataaaa
     3661 atgaatggtg acaaaatacc aacaaccatt acatcagatt cctacctaca taacggacta
     3721 agaaaaacac tacacgatgc tttaactgca aaaattcagc tcaccagttt tgaggcaaaa
     3781 tttttgagtg acatgcaaag taagtatgat ctcaatggtt cgttctcatg gctcacgcaa
     3841 aaacaacgaa ccacactaga gaacatactg gctaaatacg gaaggatctg aggttcttat
     3901 ggctcttgta tctatcagtg aagcatcaag actaacaaac aaaagtagaa caactgttca
     3961 ccgttacata tcaaagggaa aactgtccat atgcacagat gaaaacggtg taaaaaagat
     4021 agatacatca gagcttttac gagtttttgg tgcattcaaa gctgttcacc atgaacagat
     4081 cgacaatgta actactagag ctgatccttc aactcagcaa aagttcgatt tattcaacaa
     4141 agccacgttg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca
     4201 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
     4261 caacgggaaa cgtcttgctc ccgtccgcgc ttaaactcca acatggacgc tgatttatat
     4321 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     4381 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     4441 gttacagatg agatggtccg tctcaactgg ctgacggagt ttatgcctct cccgaccatc
     4501 aagcatttta tccgtactcc tgatgatgcg tggttactca ccaccgcgat tcctgggaaa
     4561 acagccttcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     4621 gccgtgttcc tgcgccggtt acattcgatt cctgtttgta attgtccttt taacagcgat
     4681 cgtgtatttc gtcttgctca ggcgcaatca cgcatgaata acggtttggt tgatgcgagt
     4741 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcacaag
     4801 ctcttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     4861 atttttgacg aggggaaatt aataggttgt attgatgttg gacgggtcgg aatcgcagac
     4921 cgttaccagg accttgccat tctttggaac tgcctcggtg agttttctcc ttcattacag
     4981 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     5041 ttgatgctcg atgagttttt ctaataatac tagcagaaat catccttagc gaaagctaag
     5101 gatttttttt atctgattac cgcctttgag tgagctctta ctgaagcacg attctactcg
     5161 gaagtggtca ataattcgtg gtttgtctgg tcaaccaccg cggtctcagt ggtgtacggt
     5221 acaaaccccc cttacggcag acctgagtaa aggagttttt tagttacctt aattgaaata
     5281 aacgaaataa aaactc
//
