LOCUS       sequence-249724-        5323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2-input history dependent program  B then A; sfGFP-0-GFP226.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5323)
  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 5323)
  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: 2SP2
FEATURES             Location/Qualifiers
     source          1..5323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(221..315)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(487..1197,1198..1200,1201..1203))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     RBS             1210..1221
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    1298..1347
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     misc_RNA        1439..1488
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             1512..1523
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1530..2243
                     /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"
     terminator      2297..2324
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    2429..2476
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     CDS             complement(2659..3474)
                     /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     3382..3401
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3466..3485
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(3832..4782)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      4830..5052
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 aatacaagaa aagcccgtca cgggcttctc agggcgtttt atggcgggtc tgctatgtgg
       61 tgctatctga ctttttgctg ttcagcagtt cctgccctct gattttccag tctgaccact
      121 tcggattatc ccgtgacagg tcattcagac tggctaatgc acccagtaag gcagcggtat
      181 catcaacagg cttacccgtc ttactgtccc tagtgcttgg attctcacca ataaaaaacg
      241 cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat
      301 ctatcaacag gagtccaagc gagctcgtaa acttggtctg acagctctag ctccggcaaa
      361 aaaacgggca aggtgtcacc accctgccct ttttctttaa aaccgaaaag attacttcgc
      421 gtttgccacc tgacgtctaa gaactcggat acccttactc tgttgaaaac gaatagatag
      481 gttttattta taaagttcgt ccataccaag tgtaatacct gctgcagtca caaactctaa
      541 tagcaccata tggtctcttt tttcgttagg gtctttcgat agggcagact gcgtcgacag
      601 gtagtggtta tccggcaaca gtaccgggcc gtcgccgatc ggtgtgttct gttggtaatg
      661 atctgcaagc tgaacgctgc catcctcgat attgtgcctg atcttgaagt taaccttaat
      721 gccgttcttc tgcttatcgg ccataatgta gacgttgtgc gagttgtagt tgtactctaa
      781 tttgtgtccg agtatattgc cgtcctcctt gaagtcaatc cccttcaact caatcctgtt
      841 cacgagcgtg tccccctcaa atttcacctc cgcccgtgtc ttgtagttcc cgtcgtcctt
      901 gaagaagatc gtcctttctt ggacgtatcc ctcaggcatg gcgcttttga agaaatcgtg
      961 ctgcttcatg tggtccgggt atctagagaa gcattggact ccgtaagtaa gagttgtcac
     1021 gagagtaggc caaggaacag gaagtttacc agtagtacaa ataaatttaa gggtcaactt
     1081 gccataagtc gcatcgccct ctccttcacc agaaacacta aatttatggc cattcacatc
     1141 accatctaac tctacaagaa taggtactac tcctgtaaaa agttcctcac ctttgctcac
     1201 catctagtat ttctcctctt tttaaacaaa attatttgta gaggggtgtt tcgtcctttc
     1261 ggactcatca gtcaaggtac gcaccttgag acacccttcg gccggcttgt cgacgacggc
     1321 ggtctccgtc gtcaggatca tccgggcttg acaattaatc atccggctcg taatgtttgt
     1381 ggaatgccaa cacaattaac atctcaatca aggtaaatgc tttttgcttt ttttgcagct
     1441 gtcaccggat gtgctttccg gtctgatgag tccgtgagga cgaaacagcc tctacaaata
     1501 attttgttta aaaagaggag aaatactaga tgcgtaaagg cgaagagctg ttcactggtg
     1561 tcgtccctat tctggtggaa ctggatggtg atgtcaacgg tcataagttt tccgtgcgtg
     1621 gcgagggtga aggtgacgca actaatggta aactgacgct gaagttcatc tgtactactg
     1681 gtaaactgcc ggtaccttgg ccgactctgg taacgacgct gacttatggt gttcagtgct
     1741 ttgctcgtta tccggaccat atgaagcagc atgacttctt caagtccgcc atgccggaag
     1801 gctatgtgca ggaacgcacg atttccttta aggatgacgg cacgtacaaa acgcgtgcgg
     1861 aagtgaaatt tgaaggcgat accctggtaa accgcattga gctgaaaggc attgacttta
     1921 aagaagacgg caatatcctg ggccataagc tggaatacaa ttttaacagc cacaatgttt
     1981 acatcaccgc cgataaacaa aaaaatggca ttaaagcgaa ttttaaaatt cgccacaacg
     2041 tggaggatgg cagcgtgcag ctggctgatc actaccagca aaacactcca atcggtgatg
     2101 gtcctgttct gctgccagac aatcactatc tgagcacgca aagcgttctg tctaaagatc
     2161 cgaacgagaa acgcgatcat atggttctgc tggagttcgt aaccgcagcg ggcatcacgc
     2221 atggtatgga tgaactgtac aaatgataaa ggcaactgaa acgattcgga tcctgtatta
     2281 ctattcttat cacactggct caccttcggg tgggcctttc tgcgtttata tactagagag
     2341 agaatataaa aagccagatt attaatccgg cttttttatt atttacttta tctgagaata
     2401 gtcaatcttc ggaaatccca ggtgtcgtgg tttgtctggt caaccaccgc ggtctcagtg
     2461 gtgtacggta caaacccact caggtctgcc gtaagggaaa ggagtttttt agttacctta
     2521 attgaaataa acgaaataaa aactcgcatt attgaccact tccgagtaga atcgtgcttc
     2581 agtaagagct cactcaaagg cggtaatcag ataaaaaaaa tccttagctt tcgctaagga
     2641 tgatttctgc tagtattatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     2701 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     2761 ctcaccgagg cagttccaaa gaatggcaag gtcctggtaa cggtctgcga ttccgacccg
     2821 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     2881 atcaccatga gtgacgactg aatccggtga gaatggcaag agcttgtgca tttctttcca
     2941 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     3001 gttattcatg cgtgattgcg cctgagcaag acgaaataca cgatcgctgt taaaaggaca
     3061 attacaaaca ggaatcgaat gtaaccggcg caggaacacg gccagcgcat caacaatatt
     3121 ttcacctgaa tcaggatatt cttctaatac ctggaaggct gttttcccag gaatcgcggt
     3181 ggtgagtaac cacgcatcat caggagtacg gataaaatgc ttgatggtcg ggagaggcat
     3241 aaactccgtc agccagttga gacggaccat ctcatctgta acatcattgg caacgctacc
     3301 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaagc gatagattgt
     3361 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcgtccat
     3421 gttggagttt aagcgcggac gggagcaaga cgtttcccgt tgaatatggc tcataacacc
     3481 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
     3541 ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac
     3601 ttttgctgag ttgaaggatc agctctagta gttacattgt cgatctgttc atggtgaaca
     3661 gctttgaatg caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt tacaccgttt
     3721 tcatctgtgc atatggacag ttttcccttt gatatgtaac ggtgaacagt tgttctactt
     3781 ttgtttgtta gtcttgatgc ttcactgata gatacaagag ccataagaac ctcagatcct
     3841 tccgtattta gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa
     3901 cgaaccattg agatcatact tactttgcat gtcactcaaa aattttgcct caaaactggt
     3961 gagctgaatt tttgcagtta aagcatcgtg tagtgttttt cttagtccgt tatgtaggta
     4021 ggaatctgat gtaatggttg ttggtatttt gtcaccattc atttttatct ggttgttctc
     4081 aagttcggtt acgagatcca tttgtctatc tagttcaact tggaaaatca acgtatcagt
     4141 cgggcggcct cgcttatcaa ccaccaattt catattgctg taagtgttta aatctttact
     4201 tattggtttc aaaacccatt ggttaagcct tttaaactca tggtagttat tttcaagcat
     4261 taacatgaac ttaaattcat caaggctaat ctctatattt gccttgtgag ttttcttttg
     4321 tgttagttct tttaataacc actcataaat cctcatagag tatttgtttt caaaagactt
     4381 aacatgttcc agattatatt ttatgaattt ttttaactgg aaaagataag gcaatatctc
     4441 ttcactaaaa actaattcta atttttcgct tgagaacttg gcatagtttg tccactggaa
     4501 aatctcaaag cctttaacca aaggattcct gatttccaca gttctcgtca tcagctctct
     4561 ggttgcttta gctaatacac cataagcatt ttccctactg atgttcatca tctgagcgta
     4621 ttggttataa gtgaacgata ccgtccgttc tttccttgta gggttttcaa tcgtggggtt
     4681 gagtagtgcc acacagcata aaattagctt ggtttcatgc tccgttaagt catagcgact
     4741 aatcgctagt tcatttgctt tgaaaacaac taattcagac atacatctca attggtctag
     4801 gtgattttaa tcactatacc aattgagatg ggctagtcaa tgataattac atgtcctttt
     4861 cctttgagtt gtgggtatct gtaaattctg ctagaccttt gctggaaaac ttgtaaattc
     4921 tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt atattcaagt
     4981 ggttataatt tatagaataa agaaagaata aaaaaagata aaaagaatag atcccagccc
     5041 tgtgtataac tcactacttt agtcagttcc gcagtattac aaaaggatgt cgcaaacgct
     5101 gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag
     5161 ctcgggcaaa tcgctgaata ttccttttgt ctccgaccat caggcacctg agtcgctgtc
     5221 tttttcgtga cattcagttc gctgcgctca cggctctggc agtgaatggg ggtaaatggc
     5281 actacaggcg ccttttatgg attcatgcaa ggaaactacc cat
//
