LOCUS       sequence-242320-        5809 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  3-input history dependent program 0-0-mKate2-sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5809)
  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 5809)
  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: 3SP4
FEATURES             Location/Qualifiers
     source          1..5809
                     /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    855..904
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     terminator      1348..1375
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             2181..2192
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2275..2322
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     terminator      2469..2500
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(2599..3414)
                     /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     3322..3341
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3406..3425
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(3772..4722)
                     /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      4770..4992
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5484..5578)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(5734..5763)
                     /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 cccttcggcc ggcttgtcga cgacggcggt ctccgtcgtc aggatcatcc
      901 gggcgggaga ccagaaacaa aaaaaggccc cccgttaggg aggccttcaa taattggttt
      961 cttttgggta tagcgtcgtg gacagtcatt catctttctg cccctccaaa agcaaaaacc
     1021 cgccgaagcg ggtttttacg taaatcaggt gaaactgacc gataagccgg gagcgccgga
     1081 tcagggagtg gacggcctgg gagcgctaca cgctgtggct gcggtcggtg ctagttgcgt
     1141 ctcagggacc cgcaaaaaaa gcaaaaagca tttaccttga ttgagatgtt aattgtgttg
     1201 gcattccaca aacattacga gccggatgat taattgtcaa cagatcaggg tgcgcaagtt
     1261 gtcaacgctc ccaggagagt tatcgacttg cgtattaggg aggcaactga aacgattcgg
     1321 atcctgtatt actattctta tcacactggc tcaccttcgg gtgggccttt ctgcgtttat
     1381 atactagaga gagaatataa aaagccagat tattaatccg gcttttttat tatttacttt
     1441 atctgagaat agtcaatctt cggaaatccc aggtgttatc aacgatgtcc taatttcgac
     1501 ggaaggtcac aataacgggc tacggctact tcatgttgtt caacgtaggt ttctttatca
     1561 gcttctttaa tacgttctaa gcgacgatca acatagtata ctccaggcat tttaaggttt
     1621 ttagcgggtt ttttggaacg ataggttgtt ttcaggttgc agattaaatg accgcctcca
     1681 actaatttca gtgccatgtc tgcgcgtcct tctaagccac cgtccgcagg atataaggtt
     1741 tctgttgatg cttcccaacc aagagttttt ttttgcatta cgggtccgtt actcgggaaa
     1801 tttacgccac ggattttaac attgtaaatc agacagccat cttgtaaact cgtatcttga
     1861 gttgctgtaa gtacaccgcc atcttcataa gttgttacac gttcccaggt aaagccttca
     1921 ggaaaggact gtttaaaaaa gtcaggaatg ccttgggtgt gattgataaa agttttagac
     1981 ccgtacataa agctcgttgc gagaatgtca aacgcaaagg gcaggggtcc accttcaact
     2041 gctttgatgc gcattgtttg tgtgccttca tatggtttac cttcaccttc ggatgtgcat
     2101 ttgaaatgat gattgttgac agtaccttcc atatataatt tcatgtgcat attttcttta
     2161 attaattctg acatctagta tttctcctct ttttaaacaa aattatttgt agaggtggtt
     2221 tcgctctgcc ttgagctcat caggtgatcg agtttcagat cacgaccact tcgtggtttg
     2281 tctggtcaac caccgcggtc tcagtggtgt acggtacaaa cccactcagg tctgccgtaa
     2341 gggaaaggag ttttttagtt accttaattg aaataaacga aataaaaact cgcattattg
     2401 accacttccg agtagaatcg tgcttcagta agagagtcac taagggttag ttagttagat
     2461 tagcagaaag tcaaaagcct ccgaccggag gcttttgact aaaacttccc ttggggttat
     2521 cattggggct cactcaaagg cggtaatcag ataaaaaaaa tccttagctt tcgctaagga
     2581 tgatttctgc tagtattatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     2641 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     2701 ctcaccgagg cagttccaaa gaatggcaag gtcctggtaa cggtctgcga ttccgacccg
     2761 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     2821 atcaccatga gtgacgactg aatccggtga gaatggcaag agcttgtgca tttctttcca
     2881 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     2941 gttattcatg cgtgattgcg cctgagcaag acgaaataca cgatcgctgt taaaaggaca
     3001 attacaaaca ggaatcgaat gtaaccggcg caggaacacg gccagcgcat caacaatatt
     3061 ttcacctgaa tcaggatatt cttctaatac ctggaaggct gttttcccag gaatcgcggt
     3121 ggtgagtaac cacgcatcat caggagtacg gataaaatgc ttgatggtcg ggagaggcat
     3181 aaactccgtc agccagttga gacggaccat ctcatctgta acatcattgg caacgctacc
     3241 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaagc gatagattgt
     3301 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcgtccat
     3361 gttggagttt aagcgcggac gggagcaaga cgtttcccgt tgaatatggc tcataacacc
     3421 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
     3481 ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac
     3541 ttttgctgag ttgaaggatc agctctagta gttacattgt cgatctgttc atggtgaaca
     3601 gctttaaatg caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt tacaccgttt
     3661 tcatctgtgc atatggacag ttttcccttt gatatctaac ggtgaacagt tgttctactt
     3721 ttgtttgtta gtcttgatgc ttcactgata gatacaagag ccataagaac ctcagatcct
     3781 tccgtattta gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa
     3841 cgaaccattg agatcatgct tactttgcat gtcactcaaa aattttgcct caaaactggt
     3901 gagctgaatt tttgcagtta aagcatcgtg tagtgttttt cttagtccgt tacgtaggta
     3961 ggaatctgat gtaatggttg ttggtatttt gtcaccattc atttttatct ggttgttctc
     4021 aagttcggtt acgagatcca tttgtctatc tagttcaact tggaaaatca acgtatcagt
     4081 cgggcggcct cgcttatcaa ccaccaattt catattgctg taagtgttta aatctttact
     4141 tattggtttc aaaacccatt ggttaagcct tttaaactca tggtagttat tttcaagcat
     4201 taacatgaac ttaaattcat caaggctaat ctctatattt gccttgtgag ttttcttttg
     4261 tgttagttct tttaataacc actcataaat cctcatagag tatttgtttt caaaagactt
     4321 aacatgttcc agattatatt ttatgaattt ttttaactgg aaaagataag gcaatatctc
     4381 ttcactaaaa actaattcta atttttcgct tgagaacttg gcatagtttg tccactggaa
     4441 aatctcaaag cctttaacca aaggattcct gatttccaca gttctcgtca tcagctctct
     4501 ggttgcttta gctaatacac cataagcatt ttccctactg atgttcatca tctgagcgta
     4561 ttggttataa gtgaacgata ccgtccgttc tttccttgta gggttttcaa tcgtggggtt
     4621 gagtagtgcc acacagcata aaattagctt ggtttcatgc tccgttaagt catagcgact
     4681 aatcgctagt tcatttgctt tgaaaacaac taattcagac atacatctca attggtctag
     4741 gtgattttaa tcactatacc aattgagatg ggctagtcaa tgataattac atgtcctttt
     4801 cctttgagtt gtgggtatct gtaaattctg ctagaccttt gctggaaaac ttgtaaattc
     4861 tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt atattcaagt
     4921 ggttataatt tatagaataa agaaagaata aaaaaagata aaaagaatag atcccagccc
     4981 tgtgtataac tcactacttt agtcagttcc gcagtattac aaaaggatgt cgcaaacgct
     5041 gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag
     5101 ctcgggcaaa tcgctgaata ttccttttgt ctccgaccat caggcacctg agtcgctgtc
     5161 tttttcgtga cattcagttc gctgcgctca cggctctggc agtgaatggg ggtaaatggc
     5221 actacaggcg ccttttatgg attcatgcaa ggaaactacc cataatacaa gaaaagcccg
     5281 tcacgggctt ctcagggcgt tttatggcgg gtctgctatg tggtgctatc tgactttttg
     5341 ctgttcagca gttcctgccc tctgattttc cagtctgacc acttcggatt atcccgtgac
     5401 aggtcattca gactggctaa tgcacccagt aaggcagcgg tatcatcaac aggcttaccc
     5461 gtcttactgt ccctagtgct tggattctca ccaataaaaa acgcccggcg gcaaccgagc
     5521 gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca
     5581 agcgagctcg taaacttggt ctgacagctc tagctccggc aaaaaaacgg gcaaggtgtc
     5641 accaccctgc cctttttctt taaaaccgaa aagattactt cgcgtttgcc acctgacgtc
     5701 taagaaaagg aatattcagc aatttgcccg tgccgaagaa aggcccaccc gtgaaggtga
     5761 gccagtgagt tgattgctac gtaattagtt agttagccct tagtgactc
//
