LOCUS       sequence-250154-        6398 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3-input history dependent program 0-BFP-mKate2-sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6398)
  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 6398)
  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: 3SP1
FEATURES             Location/Qualifiers
     source          1..6398
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             92..103
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             110..823
                     /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      998..1092
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1584..1806
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             1854..2804
                     /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(3151..3170)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3162..3977
                     /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(3235..3254)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     protein_bind    4093..4140
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     RBS             4290..4301
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      complement(5107..5134)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             5881..5892
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_RNA        complement(5916..5965)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     protein_bind    6064..6113
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
ORIGIN
        1 taattgtgtt ggcatagggt gtctcaaggt gcgtaccttg actgatgagt ccgaaaggac
       61 gaaacacccc tctacaaata attttgttta aaaagaggag aaatactaga tgcgtaaagg
      121 cgaagagctg ttcactggtg tcgtccctat tctggtggaa ctggatggtg atgtcaacgg
      181 tcataagttt tccgtgcgtg gcgagggtga aggtgacgca actaatggta aactgacgct
      241 gaagttcatc tgtactactg gtaaactgcc ggtaccttgg ccgactctgg taacgacgct
      301 gacttatggt gttcagtgct ttgctcgtta tccggaccat atgaagcagc atgacttctt
      361 caagtccgcc atgccggaag gctatgtgca ggaacgcacg atttccttta aggatgacgg
      421 cacgtacaaa acgcgtgcgg aagtgaaatt tgaaggcgat accctggtaa accgcattga
      481 gctgaaaggc attgacttta aagaagacgg caatatcctg ggccataagc tggaatacaa
      541 ttttaacagc cacaatgttt acatcaccgc cgataaacaa aaaaatggca ttaaagcgaa
      601 ttttaaaatt cgccacaacg tggaggatgg catcgtgcag ctggctgatc actaccagca
      661 aaacactcca atcggtgatg gtcctgttct gctgccagac aatcactatc tgagcacgca
      721 aagcgttctg tctaaagatc cgaacgagaa acgcgatcat atggttctgc tggagttcgt
      781 aaccgcagcg ggcatcacgc atggtatgga tgaactgtac aaatgataaa acctatctat
      841 tcgttttcaa cagagtaagg gtatccgagt tcttagacgt caggtggcaa acgcgaagta
      901 atcttttcgg ttttaaagaa aaagggcagg gtggtgacac cttgcccgtt tttttgccgg
      961 agctagagct gtcagaccaa gtttacgagc tcgcttggac tcctgttgat agatccagta
     1021 atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt
     1081 attggtgaga atccaagcac tagggacagt aagacgggta agcctgttga tgataccgct
     1141 gccttactgg gtgcattagc cagtctgaat gacctgtcac gggataatcc gaagtggtca
     1201 gactggaaaa tcagagggca ggaactgctg aacagcaaaa agtcagatag caccacatag
     1261 cagacccgcc ataaaacgcc ctgagaagcc cgtgacgggc ttttcttgta ttatgggtag
     1321 tttccttgca tgaatccata aaaggcgcct gtagtgccat ttacccccat tcactgccag
     1381 agccgtgagc gcagcgaact gaatgtcacg aaaaagacag cgactcaggt gcctgatggt
     1441 cggagacaaa aggaatattc agcgatttgc ccgagcttgc gagggtgcta cttaagcctt
     1501 tagggtttta aggtctgttt tgtagaggag caaacagcgt ttgcgacatc cttttgtaat
     1561 actgcggaac tgactaaagt agtgagttat acacagggct gggatctatt ctttttatct
     1621 ttttttattc tttctttatt ctataaatta taaccacttg aatataaaca aaaaaaacac
     1681 acaaaggtct agcggaattt acagagggtc tagcagaatt tacaagtttt ccagcaaagg
     1741 tctagcagaa tttacagata cccacaactc aaaggaaaag gacatgtaat tatcattgac
     1801 tagcccatct caattggtat agtgattaaa atcacctaga ccaattgaga tgtatgtctg
     1861 aattagttgt tttcaaagca aatgaactag cgattagtcg ctatgactta acggagcatg
     1921 aaaccaagct aattttatgc tgtgtggcac tactcaaccc cacgattgaa aaccctacaa
     1981 ggaaagaacg gacggtatcg ttcacttata accaatacgc tcagatgatg aacatcagta
     2041 gggaaaatgc ttatggtgta ttagctaaag caaccagaga gctgatgacg agaactgtgg
     2101 aaatcaggaa tcctttggtt aaaggctttg agattttcca gtggacaaac tatgccaagt
     2161 tctcaagcga aaaattagaa ttagttttta gtgaagagat attgccttat cttttccagt
     2221 taaaaaaatt cataaaatat aatctggaac atgttaagtc ttttgaaaac aaatactcta
     2281 tgaggattta tgagtggtta ttaaaagaac taacacaaaa gaaaactcac aaggcaaata
     2341 tagagattag ccttgatgaa tttaagttca tgttaatgct tgaaaataac taccatgagt
     2401 ttaaaaggct taaccaatgg gttttgaaac caataagtaa agatttaaac acttacagca
     2461 atatgaaatt ggtggttgat aagcgaggcc gcccgactga tacgttgatt ttccaagttg
     2521 aactagatag acaaatggat ctcgtaaccg aacttgagaa caaccagata aaaatgaatg
     2581 gtgacaaaat accaacaacc attacatcag attcctacct acataacgga ctaagaaaaa
     2641 cactacacga tgctttaact gcaaaaattc agctcaccag ttttgaggca aaatttttga
     2701 gtgacatgca aagtaagtat gatctcaatg gttcgttctc atggctcacg caaaaacaac
     2761 gaaccacact agagaacata ctggctaaat acggaaggat ctgaggttct tatggctctt
     2821 gtatctatca gtgaagcatc aagactaaca aacaaaagta gaacaactgt tcaccgttac
     2881 atatcaaagg gaaaactgtc catatgcaca gatgaaaacg gtgtaaaaaa gatagataca
     2941 tcagagcttt tacgagtttt tggtgcattc aaagctgttc accatgaaca gatcgacaat
     3001 gtaactacta gagctgattc ttcaactcag caaaagttcg atttattcaa caaagccacg
     3061 ttgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa
     3121 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg
     3181 aaacgtcttg ctcccgtccg cgcttaaact ccaacatgga cgctgattta tatgggtata
     3241 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc
     3301 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     3361 atgagatggt ccgtctcaac tggctgacgg agtttatgcc tctcccgacc atcaagcatt
     3421 ttatccgtac tcctgatgat gcgtggttac tcaccaccgc gattcctggg aaaacagcct
     3481 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggccgtgt
     3541 tcctgcgccg gttacattcg attcctgttt gtaattgtcc ttttaacagc gatcgtgtat
     3601 ttcgtcttgc tcaggcgcaa tcacgcatga ataacggttt ggttgatgcg agtgattttg
     3661 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcac aagctcttgc
     3721 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     3781 acgaggggaa attaataggt tgtattgatg ttggacgggt cggaatcgca gaccgttacc
     3841 aggaccttgc cattctttgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     3901 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     3961 tcgatgagtt tttctaataa tactagcaga aatcatcctt agcgaaagct aaggattttt
     4021 tttatctgat taccgccttt gagtgagctc ttactgaagc acgattctac tcggaagtgg
     4081 tcaataattc gtggtttgtc tggtcaacca ccgcggtctc agtggtgtac ggtacaaacc
     4141 ccccttacgg cagacctgag taaaggagtt ttttagttac cttaattgaa ataaacgaaa
     4201 taaaaactcg cagtggtcgt gatctgaaac tcgatcacct gatgagctca aggcagagcg
     4261 aaaccacctc tacaaataat tttgtttaaa aagaggagaa atactagatg tcagaattaa
     4321 ttaaagaaaa tatgcacatg aaattatata tggaaggtac tgtcaacaat catcatttca
     4381 aatgcacatc cgaaggtgaa ggtaaaccat atgaaggcac acaaacaatg cgcatcaaag
     4441 cagttgaagg tggacccctg ccctttgcgt ttgacattct cgcaacgagc tttatgtacg
     4501 ggtctaaaac ttttatcaat cacacccaag gcattcctga cttttttaaa cagtcctttc
     4561 ctgaaggctt tacctgggaa cgtgtaacaa cttatgaaga tggcggtgta cttacagcaa
     4621 ctcaagatac gagtttacaa gatggctgtc tgatttacaa tgttaaaatc cgtggcgtaa
     4681 atttcccgag taacggaccc gtaatgcaaa aaaaaactct tggttgggaa gcatcaacag
     4741 aaaccttata tcctgcggac ggtggcttag aaggacgcgc agacatggca ctgaaattag
     4801 ttggaggcgg tcatttaatc tgcaacctga aaacaaccta tcgttccaaa aaacccgcta
     4861 aaaaccttaa aatgcctgga gtatactatg ttgatcgtcg cttagaacgt attaaagaag
     4921 ctgataaaga aacctacgtt gaacaacatg aagtagccgt agcccgttat tgtgaccttc
     4981 cgtcgaaatt aggacatcgt tgataacacc tgggatttcc gaagattgac tattctcaga
     5041 taaagtaaat aataaaaaag ccggattaat aatctggctt tttatattct ctctctagta
     5101 tataaacgca gaaaggccca cccgaaggtg agccagtgtg ataagaatag taatacagga
     5161 tccgaatcgt ttcagttgcc tttagtgccc cagtttgcta gggaggtcgc agtatctggc
     5221 cactgccacc tcgtgctgct cgacgtaggt ctcgttgttg gcctccttga ttctttccag
     5281 tctgtagtcc acatagtaga cgccaggcat cttgaggttc ttagcgggtt tcttggatct
     5341 atatgtggtc ttgatgtttg cgatcagatg gctcccgccc acgagcttca gggccatgtc
     5401 gtttctgcct tccaggccgc cgtcagcggg gtacagcgtc tcggtgaagg cctcccagcc
     5461 gagtgttttc ttctgcatca cagggccgtt ggatgtgaag ttcacccctc tgatcttgac
     5521 gttgtagatg aggcagccgt cctggaggct ggtgtcctgg gtagcggtca gcacgccccc
     5581 gtcttcgtat gtggtgactc tctcccatgt gaagccctca gggaaggact gcttgaagaa
     5641 gtcggggatg ccctgggtgt ggttgatgaa ggtcttgctg ccgtagagga agctagtagc
     5701 caggatgtcg aaggcgaagg ggagagggcc gccctcgacc accttgattc tcatggtctg
     5761 ggtgccctcg tagggcttgc cttcgccctc ggatgtgcac ttgaagtgat ggttgtccac
     5821 ggtgccctcc atgtacagct tcatgtgcat gttctcctta atcagctcgc tcatctagta
     5881 tttctcctct ttttaaacaa aattatttgt agaggctgtt tcgtcctcac ggactcatca
     5941 gaccggaaag cacatccggt gacagctccc taatacgcaa gtagataact ctcctgggag
     6001 cgttgacaac ttgcgcaccc tgatctgttg acaattaatc atccggctcg taatgtttgt
     6061 ggatcggccg gcttgtcgac gacggcggtc tccgtcgtca ggatcatccg ggcgggtccc
     6121 tgagacgcaa ctagcaccga ccgcagccac agcgtgtagc gctcccaggc cgtccactcc
     6181 ctgatccggc gctcccggct tatcggtcag tttcacctga tttacgtaaa aacccgcttc
     6241 ggcgggtttt tgcttttgga ggggcagaaa gatgaatgac tgtccacgac gctataccca
     6301 aaagaaacca attattgaag gcctccctaa cggggggcct ttttttgttt ctggtctccc
     6361 gcaaaaaaag caaaaagcat ttaccttgat tgagatgt
//
