LOCUS       sequence-250156-        5603 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 5603)
  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 5603)
  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..5603
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      262..484
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             532..1482
                     /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(1829..1848)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1840..2655
                     /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(1913..1932)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     protein_bind    2771..2818
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     terminator      complement(2990..3017)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             3764..3775
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_RNA        complement(3799..3848)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     protein_bind    3947..3996
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     RBS             4373..4384
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4391..5104
                     /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
                     NFKIRHNVEDGIVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      5279..5373
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 tccttgcatg aatccataaa aggcgcctgt agtgccattt acccccattc actgccagag
       61 ccgtgagcgc agcgaactga atgtcacgaa aaagacagcg actcaggtgc ctgatggtcg
      121 gagacaaaag gaatattcag cgatttgccc gagcttgcga gggtgctact taagccttta
      181 gggttttaag gtctgttttg tagaggagca aacagcgttt gcgacatcct tttgtaatac
      241 tgcggaactg actaaagtag tgagttatac acagggctgg gatctattct ttttatcttt
      301 ttttattctt tctttattct ataaattata accacttgaa tataaacaaa aaaaacacac
      361 aaaggtctag cggaatttac agagggtcta gcagaattta caagttttcc agcaaaggtc
      421 tagcagaatt tacagatacc cacaactcaa aggaaaagga catgtaatta tcattgacta
      481 gcccatctca attggtatag tgattaaaat cacctagacc aattgagatg tatgtctgaa
      541 ttagttgttt tcaaagcaaa tgaactagcg attagtcgct atgacttaac ggagcatgaa
      601 accaagctaa ttttatgctg tgtggcacta ctcaacccca cgattgaaaa ccctacaagg
      661 aaagaacgga cggtatcgtt cacttataac caatacgctc agatgatgaa catcagtagg
      721 gaaaatgctt atggtgtatt agctaaagca accagagagc tgatgacgag aactgtggaa
      781 atcaggaatc ctttggttaa aggctttgag attttccagt ggacaaacta tgccaagttc
      841 tcaagcgaaa aattagaatt agtttttagt gaagagatat tgccttatct tttccagtta
      901 aaaaaattca taaaatataa tctggaacat gttaagtctt ttgaaaacaa atactctatg
      961 aggatttatg agtggttatt aaaagaacta acacaaaaga aaactcacaa ggcaaatata
     1021 gagattagcc ttgatgaatt taagttcatg ttaatgcttg aaaataacta ccatgagttt
     1081 aaaaggctta accaatgggt tttgaaacca ataagtaaag atttaaacac ttacagcaat
     1141 atgaaattgg tggttgataa gcgaggccgc ccgactgata cgttgatttt ccaagttgaa
     1201 ctagatagac aaatggatct cgtaaccgaa cttgagaaca accagataaa aatgaatggt
     1261 gacaaaatac caacaaccat tacatcagat tcctacctac ataacggact aagaaaaaca
     1321 ctacacgatg ctttaactgc aaaaattcag ctcaccagtt ttgaggcaaa atttttgagt
     1381 gacatgcaaa gtaagtatga tctcaatggt tcgttctcat ggctcacgca aaaacaacga
     1441 accacactag agaacatact ggctaaatac ggaaggatct gaggttctta tggctcttgt
     1501 atctatcagt gaagcatcaa gactaacaaa caaaagtaga acaactgttc accgttacat
     1561 atcaaaggga aaactgtcca tatgcacaga tgaaaacggt gtaaaaaaga tagatacatc
     1621 agagctttta cgagtttttg gtgcattcaa agctgttcac catgaacaga tcgacaatgt
     1681 aactactaga gctgattctt caactcagca aaagttcgat ttattcaaca aagccacgtt
     1741 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     1801 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     1861 acgtcttgct cccgtccgcg cttaaactcc aacatggacg ctgatttata tgggtataaa
     1921 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
     1981 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     2041 gagatggtcc gtctcaactg gctgacggag tttatgcctc tcccgaccat caagcatttt
     2101 atccgtactc ctgatgatgc gtggttactc accaccgcga ttcctgggaa aacagccttc
     2161 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggccgtgttc
     2221 ctgcgccggt tacattcgat tcctgtttgt aattgtcctt ttaacagcga tcgtgtattt
     2281 cgtcttgctc aggcgcaatc acgcatgaat aacggtttgg ttgatgcgag tgattttgat
     2341 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcacaa gctcttgcca
     2401 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     2461 gaggggaaat taataggttg tattgatgtt ggacgggtcg gaatcgcaga ccgttaccag
     2521 gaccttgcca ttctttggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     2581 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     2641 gatgagtttt tctaataata ctagcagaaa tcatccttag cgaaagctaa ggattttttt
     2701 tatctgatta ccgcctttga gtgagctctt actgaagcac gattctactc ggaagtggtc
     2761 aataattcgt ggtttgtctg gtcaaccacc gcggtctcag tggtgtacgg tacaaacccc
     2821 ccttacggca gacctgagta aaggagtttt ttagttacct taattgaaat aaacgaaata
     2881 aaaactcgcc acctgggatt tccgaagatt gactattctc agataaagta aataataaaa
     2941 aagccggatt aataatctgg ctttttatat tctctctcta gtatataaac gcagaaaggc
     3001 ccacccgaag gtgagccagt gtgataagaa tagtaataca ggatccgaat cgtttcagtt
     3061 gcctttagtg ccccagtttg ctagggaggt cgcagtatct ggccactgcc acctcgtgct
     3121 gctcgacgta ggtctcgttg ttggcctcct tgattctttc cagtctgtag tccacatagt
     3181 agacgccagg catcttgagg ttcttagcgg gtttcttgga tctatatgtg gtcttgatgt
     3241 ttgcgatcag atggctcccg cccacgagct tcagggccat gtcgtttctg ccttccaggc
     3301 cgccgtcagc ggggtacagc gtctcggtga aggcctccca gccgagtgtt ttcttctgca
     3361 tcacagggcc gttggatgtg aagttcaccc ctctgatctt gacgttgtag atgaggcagc
     3421 cgtcctggag gctggtgtcc tgggtagcgg tcagcacgcc cccgtcttcg tatgtggtga
     3481 ctctctccca tgtgaagccc tcagggaagg actgcttgaa gaagtcgggg atgccctggg
     3541 tgtggttgat gaaggtcttg ctgccgtaga ggaagctagt agccaggatg tcgaaggcga
     3601 aggggagagg gccgccctcg accaccttga ttctcatggt ctgggtgccc tcgtagggct
     3661 tgccttcgcc ctcggatgtg cacttgaagt gatggttgtc cacggtgccc tccatgtaca
     3721 gcttcatgtg catgttctcc ttaatcagct cgctcatcta gtatttctcc tctttttaaa
     3781 caaaattatt tgtagaggct gtttcgtcct cacggactca tcagaccgga aagcacatcc
     3841 ggtgacagct ccctaatacg caagtagata actctcctgg gagcgttgac aacttgcgca
     3901 ccctgatctg ttgacaatta atcatccggc tcgtaatgtt tgtggatcgg ccggcttgtc
     3961 gacgacggcg gtctccgtcg tcaggatcat ccgggcgggt ccctgagacg caactagcac
     4021 cgaccgcagc cacagcgtgt agcgctccca ggccgtccac tccctgatcc ggcgctcccg
     4081 gcttatcggt cagtttcacc tgatttacgt aaaaacccgc ttcggcgggt ttttgctttt
     4141 ggaggggcag aaagatgaat gactgtccac gacgctatac ccaaaagaaa ccaattattg
     4201 aaggcctccc taacgggggg cctttttttg tttctggtct cccgcaaaaa aagcaaaaag
     4261 catttacctt gattgagatg ttaattgtgt tggcataggg tgtctcaagg tgcgtacctt
     4321 gactgatgag tccgaaagga cgaaacaccc ctctacaaat aattttgttt aaaaagagga
     4381 gaaatactag atgcgtaaag gcgaagagct gttcactggt gtcgtcccta ttctggtgga
     4441 actggatggt gatgtcaacg gtcataagtt ttccgtgcgt ggcgagggtg aaggtgacgc
     4501 aactaatggt aaactgacgc tgaagttcat ctgtactact ggtaaactgc cggtaccttg
     4561 gccgactctg gtaacgacgc tgacttatgg tgttcagtgc tttgctcgtt atccggacca
     4621 tatgaagcag catgacttct tcaagtccgc catgccggaa ggctatgtgc aggaacgcac
     4681 gatttccttt aaggatgacg gcacgtacaa aacgcgtgcg gaagtgaaat ttgaaggcga
     4741 taccctggta aaccgcattg agctgaaagg cattgacttt aaagaagacg gcaatatcct
     4801 gggccataag ctggaataca attttaacag ccacaatgtt tacatcaccg ccgataaaca
     4861 aaaaaatggc attaaagcga attttaaaat tcgccacaac gtggaggatg gcatcgtgca
     4921 gctggctgat cactaccagc aaaacactcc aatcggtgat ggtcctgttc tgctgccaga
     4981 caatcactat ctgagcacgc aaagcgttct gtctaaagat ccgaacgaga aacgcgatca
     5041 tatggttctg ctggagttcg taaccgcagc gggcatcacg catggtatgg atgaactgta
     5101 caaatgataa aacctatcta ttcgttttca acagagtaag ggtatccgag ttcttagacg
     5161 tcaggtggca aacgcgaagt aatcttttcg gttttaaaga aaaagggcag ggtggtgaca
     5221 ccttgcccgt ttttttgccg gagctagagc tgtcagacca agtttacgag ctcgcttgga
     5281 ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc
     5341 ggttgccgcc gggcgttttt tattggtgag aatccaagca ctagggacag taagacgggt
     5401 aagcctgttg atgataccgc tgccttactg ggtgcattag ccagtctgaa tgacctgtca
     5461 cgggataatc cgaagtggtc agactggaaa atcagagggc aggaactgct gaacagcaaa
     5521 aagtcagata gcaccacata gcagacccgc cataaaacgc cctgagaagc ccgtgacggg
     5581 cttttcttgt attatgggta gtt
//
