LOCUS       sequence-242328-        5493 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  2-input history dependent program A then B;BFP-0-sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5493)
  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 5493)
  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..5493
                     /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"
     protein_bind    2026..2073
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     terminator      2153..2184
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(2283..3098)
                     /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     3006..3025
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3090..3109
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(3456..4406)
                     /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      4454..4676
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5168..5262)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(5418..5447)
                     /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 ggtggcgagt ttttatttcg tttatttcaa
     1981 ttaaggtaac taaaaaactc ctttactcag gtctgccgta aggggggttt gtaccgtaca
     2041 ccactgagac cgcggtggtt gaccagacaa accacgaatt attgaccact tccgagtaga
     2101 atcgtgcttc agtaagagag tcactaaggg ttagttagtt agattagcag aaagtcaaaa
     2161 gcctccgacc ggaggctttt gactaaaact tcccttgggg ttatcattgg ggctcactca
     2221 aaggcggtaa tcagataaaa aaaatcctta gctttcgcta aggatgattt ctgctagtat
     2281 tattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     2341 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     2401 caaagaatgg caaggtcctg gtaacggtct gcgattccga cccgtccaac atcaatacaa
     2461 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     2521 actgaatccg gtgagaatgg caagagcttg tgcatttctt tccagacttg ttcaacaggc
     2581 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt catgcgtgat
     2641 tgcgcctgag caagacgaaa tacacgatcg ctgttaaaag gacaattaca aacaggaatc
     2701 gaatgtaacc ggcgcaggaa cacggccagc gcatcaacaa tattttcacc tgaatcagga
     2761 tattcttcta atacctggaa ggctgttttc ccaggaatcg cggtggtgag taaccacgca
     2821 tcatcaggag tacggataaa atgcttgatg gtcgggagag gcataaactc cgtcagccag
     2881 ttgagacgga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     2941 aacaactctg gcgcatcggg cttcccatac aagcgataga ttgtcgcacc tgattgcccg
     3001 acattatcgc gagcccattt atacccatat aaatcagcgt ccatgttgga gtttaagcgc
     3061 ggacgggagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     3121 atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat
     3181 cagagatttt gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag
     3241 gatcagctct agtagttaca ttgtcgatct gttcatggtg aacagcttta aatgcaccaa
     3301 aaactcgtaa aagctctgat gtatctatct tttttacacc gttttcatct gtgcatatgg
     3361 acagttttcc ctttgatatc taacggtgaa cagttgttct acttttgttt gttagtcttg
     3421 atgcttcact gatagataca agagccataa gaacctcaga tccttccgta tttagccagt
     3481 atgttctcta gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc attgagatca
     3541 tgcttacttt gcatgtcact caaaaatttt gcctcaaaac tggtgagctg aatttttgca
     3601 gttaaagcat cgtgtagtgt ttttcttagt ccgttacgta ggtaggaatc tgatgtaatg
     3661 gttgttggta ttttgtcacc attcattttt atctggttgt tctcaagttc ggttacgaga
     3721 tccatttgtc tatctagttc aacttggaaa atcaacgtat cagtcgggcg gcctcgctta
     3781 tcaaccacca atttcatatt gctgtaagtg tttaaatctt tacttattgg tttcaaaacc
     3841 cattggttaa gccttttaaa ctcatggtag ttattttcaa gcattaacat gaacttaaat
     3901 tcatcaaggc taatctctat atttgccttg tgagttttct tttgtgttag ttcttttaat
     3961 aaccactcat aaatcctcat agagtatttg ttttcaaaag acttaacatg ttccagatta
     4021 tattttatga atttttttaa ctggaaaaga taaggcaata tctcttcact aaaaactaat
     4081 tctaattttt cgcttgagaa cttggcatag tttgtccact ggaaaatctc aaagccttta
     4141 accaaaggat tcctgatttc cacagttctc gtcatcagct ctctggttgc tttagctaat
     4201 acaccataag cattttccct actgatgttc atcatctgag cgtattggtt ataagtgaac
     4261 gataccgtcc gttctttcct tgtagggttt tcaatcgtgg ggttgagtag tgccacacag
     4321 cataaaatta gcttggtttc atgctccgtt aagtcatagc gactaatcgc tagttcattt
     4381 gctttgaaaa caactaattc agacatacat ctcaattggt ctaggtgatt ttaatcacta
     4441 taccaattga gatgggctag tcaatgataa ttacatgtcc ttttcctttg agttgtgggt
     4501 atctgtaaat tctgctagac ctttgctgga aaacttgtaa attctgctag accctctgta
     4561 aattccgcta gacctttgtg tgtttttttt gtttatattc aagtggttat aatttataga
     4621 ataaagaaag aataaaaaaa gataaaaaga atagatccca gccctgtgta taactcacta
     4681 ctttagtcag ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa
     4741 aacagacctt aaaaccctaa aggcttaagt agcaccctcg caagctcggg caaatcgctg
     4801 aatattcctt ttgtctccga ccatcaggca cctgagtcgc tgtctttttc gtgacattca
     4861 gttcgctgcg ctcacggctc tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt
     4921 atggattcat gcaaggaaac tacccataat acaagaaaag cccgtcacgg gcttctcagg
     4981 gcgttttatg gcgggtctgc tatgtggtgc tatctgactt tttgctgttc agcagttcct
     5041 gccctctgat tttccagtct gaccacttcg gattatcccg tgacaggtca ttcagactgg
     5101 ctaatgcacc cagtaaggca gcggtatcat caacaggctt acccgtctta ctgtccctag
     5161 tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca
     5221 gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag ctcgtaaact
     5281 tggtctgaca gctctagctc cggcaaaaaa acgggcaagg tgtcaccacc ctgccctttt
     5341 tctttaaaac cgaaaagatt acttcgcgtt tgccacctga cgtctaagaa aaggaatatt
     5401 cagcaatttg cccgtgccga agaaaggccc acccgtgaag gtgagccagt gagttgattg
     5461 ctacgtaatt agttagttag cccttagtga ctc
//
