LOCUS       sequence-250997-        5707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3-input history dependent program 0-BFP-0-sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5707)
  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 5707)
  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: 3SP5
FEATURES             Location/Qualifiers
     source          1..5707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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    1155..1204
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     misc_RNA        1303..1352
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             1376..1387
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2134..2161
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    2333..2380
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     CDS             complement(2497..3312)
                     /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     3220..3239
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3304..3323
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(3670..4620)
                     /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      4668..4890
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5382..5476)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(5632..5661)
                     /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 tttttgcggg agaccagaaa caaaaaaagg ccccccgtta gggaggcctt caataattgg
      961 tttcttttgg gtatagcgtc gtggacagtc attcatcttt ctgcccctcc aaaagcaaaa
     1021 acccgccgaa gcgggttttt acgtaaatca ggtgaaactg accgataagc cgggagcgcc
     1081 ggatcaggga gtggacggcc tgggagcgct acacgctgtg gctgcggtcg gtgctagttg
     1141 cgtctcaggg acccgcccgg atgatcctga cgacggagac cgccgtcgtc gacaagccgg
     1201 ccgatccaca aacattacga gccggatgat taattgtcaa cagatcaggg tgcgcaagtt
     1261 gtcaacgctc ccaggagagt tatcgacttg cgtattaggg agctgtcacc ggatgtgctt
     1321 tccggtctga tgagtccgtg aggacgaaac agcctctaca aataattttg tttaaaaaga
     1381 ggagaaatac tagatgagcg agctgattaa ggagaacatg cacatgaagc tgtacatgga
     1441 gggcaccgtg gacaaccatc acttcaagtg cacatccgag ggcgaaggca agccctacga
     1501 gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct tcgccttcga
     1561 catcctggct actagcttcc tctacggcag caagaccttc atcaaccaca cccagggcat
     1621 ccccgacttc ttcaagcagt ccttccctga gggcttcaca tgggagagag tcaccacata
     1681 cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat
     1741 ctacaacgtc aagatcagag gggtgaactt cacatccaac ggccctgtga tgcagaagaa
     1801 aacactcggc tgggaggcct tcaccgagac gctgtacccc gctgacggcg gcctggaagg
     1861 cagaaacgac atggccctga agctcgtggg cgggagccat ctgatcgcaa acatcaagac
     1921 cacatataga tccaagaaac ccgctaagaa cctcaagatg cctggcgtct actatgtgga
     1981 ctacagactg gaaagaatca aggaggccaa caacgagacc tacgtcgagc agcacgaggt
     2041 ggcagtggcc agatactgcg acctccctag caaactgggg cactaaaggc aactgaaacg
     2101 attcggatcc tgtattacta ttcttatcac actggctcac cttcgggtgg gcctttctgc
     2161 gtttatatac tagagagaga atataaaaag ccagattatt aatccggctt ttttattatt
     2221 tactttatct gagaatagtc aatcttcgga aatcccaggt ggcgagtttt tatttcgttt
     2281 atttcaatta aggtaactaa aaaactcctt tactcaggtc tgccgtaagg ggggtttgta
     2341 ccgtacacca ctgagaccgc ggtggttgac cagacaaacc acgaattatt gaccacttcc
     2401 gagtagaatc gtgcttcagt aagaggctca ctcaaaggcg gtaatcagat aaaaaaaatc
     2461 cttagctttc gctaaggatg atttctgcta gtattattag aaaaactcat cgagcatcaa
     2521 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     2581 ctgtaatgaa ggagaaaact caccgaggca gttccaaaga atggcaaggt cctggtaacg
     2641 gtctgcgatt ccgacccgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     2701 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaagag
     2761 cttgtgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     2821 actcgcatca accaaaccgt tattcatgcg tgattgcgcc tgagcaagac gaaatacacg
     2881 atcgctgtta aaaggacaat tacaaacagg aatcgaatgt aaccggcgca ggaacacggc
     2941 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaaggctgt
     3001 tttcccagga atcgcggtgg tgagtaacca cgcatcatca ggagtacgga taaaatgctt
     3061 gatggtcggg agaggcataa actccgtcag ccagttgaga cggaccatct catctgtaac
     3121 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     3181 atacaagcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     3241 atataaatca gcgtccatgt tggagtttaa gcgcggacgg gagcaagacg tttcccgttg
     3301 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     3361 tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtggct
     3421 ttgttgaata aatcgaactt ttgctgagtt gaaggatcag ctctagtagt tacattgtcg
     3481 atctgttcat ggtgaacagc tttaaatgca ccaaaaactc gtaaaagctc tgatgtatct
     3541 atctttttta caccgttttc atctgtgcat atggacagtt ttccctttga tatctaacgg
     3601 tgaacagttg ttctactttt gtttgttagt cttgatgctt cactgataga tacaagagcc
     3661 ataagaacct cagatccttc cgtatttagc cagtatgttc tctagtgtgg ttcgttgttt
     3721 ttgcgtgagc catgagaacg aaccattgag atcatgctta ctttgcatgt cactcaaaaa
     3781 ttttgcctca aaactggtga gctgaatttt tgcagttaaa gcatcgtgta gtgtttttct
     3841 tagtccgtta cgtaggtagg aatctgatgt aatggttgtt ggtattttgt caccattcat
     3901 ttttatctgg ttgttctcaa gttcggttac gagatccatt tgtctatcta gttcaacttg
     3961 gaaaatcaac gtatcagtcg ggcggcctcg cttatcaacc accaatttca tattgctgta
     4021 agtgtttaaa tctttactta ttggtttcaa aacccattgg ttaagccttt taaactcatg
     4081 gtagttattt tcaagcatta acatgaactt aaattcatca aggctaatct ctatatttgc
     4141 cttgtgagtt ttcttttgtg ttagttcttt taataaccac tcataaatcc tcatagagta
     4201 tttgttttca aaagacttaa catgttccag attatatttt atgaattttt ttaactggaa
     4261 aagataaggc aatatctctt cactaaaaac taattctaat ttttcgcttg agaacttggc
     4321 atagtttgtc cactggaaaa tctcaaagcc tttaaccaaa ggattcctga tttccacagt
     4381 tctcgtcatc agctctctgg ttgctttagc taatacacca taagcatttt ccctactgat
     4441 gttcatcatc tgagcgtatt ggttataagt gaacgatacc gtccgttctt tccttgtagg
     4501 gttttcaatc gtggggttga gtagtgccac acagcataaa attagcttgg tttcatgctc
     4561 cgttaagtca tagcgactaa tcgctagttc atttgctttg aaaacaacta attcagacat
     4621 acatctcaat tggtctaggt gattttaatc actataccaa ttgagatggg ctagtcaatg
     4681 ataattacat gtccttttcc tttgagttgt gggtatctgt aaattctgct agacctttgc
     4741 tggaaaactt gtaaattctg ctagaccctc tgtaaattcc gctagacctt tgtgtgtttt
     4801 ttttgtttat attcaagtgg ttataattta tagaataaag aaagaataaa aaaagataaa
     4861 aagaatagat cccagccctg tgtataactc actactttag tcagttccgc agtattacaa
     4921 aaggatgtcg caaacgctgt ttgctcctct acaaaacaga ccttaaaacc ctaaaggctt
     4981 aagtagcacc ctcgcaagct cgggcaaatc gctgaatatt ccttttgtct ccgaccatca
     5041 ggcacctgag tcgctgtctt tttcgtgaca ttcagttcgc tgcgctcacg gctctggcag
     5101 tgaatggggg taaatggcac tacaggcgcc ttttatggat tcatgcaagg aaactaccca
     5161 taatacaaga aaagcccgtc acgggcttct cagggcgttt tatggcgggt ctgctatgtg
     5221 gtgctatctg actttttgct gttcagcagt tcctgccctc tgattttcca gtctgaccac
     5281 ttcggattat cccgtgacag gtcattcaga ctggctaatg cacccagtaa ggcagcggta
     5341 tcatcaacag gcttacccgt cttactgtcc ctagtgcttg gattctcacc aataaaaaac
     5401 gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga
     5461 tctatcaaca ggagtccaag cgagctcgta aacttggtct gacagctcta gctccggcaa
     5521 aaaaacgggc aaggtgtcac caccctgccc tttttcttta aaaccgaaaa gattacttcg
     5581 cgtttgccac ctgacgtcta agaaaaggaa tattcagcaa tttgcccgtg ccgaagaaag
     5641 gcccacccgt gaaggtgagc cagtgagttg attgctacgt aattagttag ttagccctta
     5701 gtgactc
//
