LOCUS       sequence-242330-        5502 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  2-input history dependent program A then B;0-mKate2-sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5502)
  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 5502)
  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: 2SP7
FEATURES             Location/Qualifiers
     source          1..5502
                     /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)"
     terminator      1041..1068
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1874..1885
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2035..2082
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     terminator      2162..2193
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(2292..3107)
                     /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     3015..3034
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3099..3118
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(3465..4415)
                     /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      4463..4685
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5177..5271)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(5427..5456)
                     /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 ggcaggcaac tgaaacgatt cggatcctgt
     1021 attactattc ttatcacact ggctcacctt cgggtgggcc tttctgcgtt tatatactag
     1081 agagagaata taaaaagcca gattattaat ccggcttttt tattatttac tttatctgag
     1141 aatagtcaat cttcggaaat cccaggtgtt atcaacgatg tcctaatttc gacggaaggt
     1201 cacaataacg ggctacggct acttcatgtt gttcaacgta ggtttcttta tcagcttctt
     1261 taatacgttc taagcgacga tcaacatagt atactccagg cattttaagg tttttagcgg
     1321 gttttttgga acgataggtt gttttcaggt tgcagattaa atgaccgcct ccaactaatt
     1381 tcagtgccat gtctgcgcgt ccttctaagc caccgtccgc aggatataag gtttctgttg
     1441 atgcttccca accaagagtt tttttttgca ttacgggtcc gttactcggg aaatttacgc
     1501 cacggatttt aacattgtaa atcagacagc catcttgtaa actcgtatct tgagttgctg
     1561 taagtacacc gccatcttca taagttgtta cacgttccca ggtaaagcct tcaggaaagg
     1621 actgtttaaa aaagtcagga atgccttggg tgtgattgat aaaagtttta gacccgtaca
     1681 taaagctcgt tgcgagaatg tcaaacgcaa agggcagggg tccaccttca actgctttga
     1741 tgcgcattgt ttgtgtgcct tcatatggtt taccttcacc ttcggatgtg catttgaaat
     1801 gatgattgtt gacagtacct tccatatata atttcatgtg catattttct ttaattaatt
     1861 ctgacatcta gtatttctcc tctttttaaa caaaattatt tgtagaggtg gtttcgctct
     1921 gccttgagct catcaggtga tcgagtttca gatcacgacc actgcgagtt tttatttcgt
     1981 ttatttcaat taaggtaact aaaaaactcc tttactcagg tctgccgtaa ggggggtttg
     2041 taccgtacac cactgagacc gcggtggttg accagacaaa ccacgaatta ttgaccactt
     2101 ccgagtagaa tcgtgcttca gtaagagagt cactaagggt tagttagtta gattagcaga
     2161 aagtcaaaag cctccgaccg gaggcttttg actaaaactt cccttggggt tatcattggg
     2221 gctcactcaa aggcggtaat cagataaaaa aaatccttag ctttcgctaa ggatgatttc
     2281 tgctagtatt attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     2341 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     2401 aggcagttcc aaagaatggc aaggtcctgg taacggtctg cgattccgac ccgtccaaca
     2461 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     2521 tgagtgacga ctgaatccgg tgagaatggc aagagcttgt gcatttcttt ccagacttgt
     2581 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     2641 atgcgtgatt gcgcctgagc aagacgaaat acacgatcgc tgttaaaagg acaattacaa
     2701 acaggaatcg aatgtaaccg gcgcaggaac acggccagcg catcaacaat attttcacct
     2761 gaatcaggat attcttctaa tacctggaag gctgttttcc caggaatcgc ggtggtgagt
     2821 aaccacgcat catcaggagt acggataaaa tgcttgatgg tcgggagagg cataaactcc
     2881 gtcagccagt tgagacggac catctcatct gtaacatcat tggcaacgct acctttgcca
     2941 tgtttcagaa acaactctgg cgcatcgggc ttcccataca agcgatagat tgtcgcacct
     3001 gattgcccga cattatcgcg agcccattta tacccatata aatcagcgtc catgttggag
     3061 tttaagcgcg gacgggagca agacgtttcc cgttgaatat ggctcataac accccttgta
     3121 ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca
     3181 atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct
     3241 gagttgaagg atcagctcta gtagttacat tgtcgatctg ttcatggtga acagctttaa
     3301 atgcaccaaa aactcgtaaa agctctgatg tatctatctt ttttacaccg ttttcatctg
     3361 tgcatatgga cagttttccc tttgatatct aacggtgaac agttgttcta cttttgtttg
     3421 ttagtcttga tgcttcactg atagatacaa gagccataag aacctcagat ccttccgtat
     3481 ttagccagta tgttctctag tgtggttcgt tgtttttgcg tgagccatga gaacgaacca
     3541 ttgagatcat gcttactttg catgtcactc aaaaattttg cctcaaaact ggtgagctga
     3601 atttttgcag ttaaagcatc gtgtagtgtt tttcttagtc cgttacgtag gtaggaatct
     3661 gatgtaatgg ttgttggtat tttgtcacca ttcattttta tctggttgtt ctcaagttcg
     3721 gttacgagat ccatttgtct atctagttca acttggaaaa tcaacgtatc agtcgggcgg
     3781 cctcgcttat caaccaccaa tttcatattg ctgtaagtgt ttaaatcttt acttattggt
     3841 ttcaaaaccc attggttaag ccttttaaac tcatggtagt tattttcaag cattaacatg
     3901 aacttaaatt catcaaggct aatctctata tttgccttgt gagttttctt ttgtgttagt
     3961 tcttttaata accactcata aatcctcata gagtatttgt tttcaaaaga cttaacatgt
     4021 tccagattat attttatgaa tttttttaac tggaaaagat aaggcaatat ctcttcacta
     4081 aaaactaatt ctaatttttc gcttgagaac ttggcatagt ttgtccactg gaaaatctca
     4141 aagcctttaa ccaaaggatt cctgatttcc acagttctcg tcatcagctc tctggttgct
     4201 ttagctaata caccataagc attttcccta ctgatgttca tcatctgagc gtattggtta
     4261 taagtgaacg ataccgtccg ttctttcctt gtagggtttt caatcgtggg gttgagtagt
     4321 gccacacagc ataaaattag cttggtttca tgctccgtta agtcatagcg actaatcgct
     4381 agttcatttg ctttgaaaac aactaattca gacatacatc tcaattggtc taggtgattt
     4441 taatcactat accaattgag atgggctagt caatgataat tacatgtcct tttcctttga
     4501 gttgtgggta tctgtaaatt ctgctagacc tttgctggaa aacttgtaaa ttctgctaga
     4561 ccctctgtaa attccgctag acctttgtgt gttttttttg tttatattca agtggttata
     4621 atttatagaa taaagaaaga ataaaaaaag ataaaaagaa tagatcccag ccctgtgtat
     4681 aactcactac tttagtcagt tccgcagtat tacaaaagga tgtcgcaaac gctgtttgct
     4741 cctctacaaa acagacctta aaaccctaaa ggcttaagta gcaccctcgc aagctcgggc
     4801 aaatcgctga atattccttt tgtctccgac catcaggcac ctgagtcgct gtctttttcg
     4861 tgacattcag ttcgctgcgc tcacggctct ggcagtgaat gggggtaaat ggcactacag
     4921 gcgcctttta tggattcatg caaggaaact acccataata caagaaaagc ccgtcacggg
     4981 cttctcaggg cgttttatgg cgggtctgct atgtggtgct atctgacttt ttgctgttca
     5041 gcagttcctg ccctctgatt ttccagtctg accacttcgg attatcccgt gacaggtcat
     5101 tcagactggc taatgcaccc agtaaggcag cggtatcatc aacaggctta cccgtcttac
     5161 tgtccctagt gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga
     5221 acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc
     5281 tcgtaaactt ggtctgacag ctctagctcc ggcaaaaaaa cgggcaaggt gtcaccaccc
     5341 tgcccttttt ctttaaaacc gaaaagatta cttcgcgttt gccacctgac gtctaagaaa
     5401 aggaatattc agcaatttgc ccgtgccgaa gaaaggccca cccgtgaagg tgagccagtg
     5461 agttgattgc tacgtaatta gttagttagc ccttagtgac tc
//
