LOCUS       sequence-246180-        6091 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2-input history dependent program A then B;BFP-mKate2-sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6091)
  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 6091)
  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: 2SP9
FEATURES             Location/Qualifiers
     source          1..6091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(175..269)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(444..1157)
                     /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             1164..1175
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    1341..1390
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     misc_RNA        1393..1442
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             1466..1477
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2224..2251
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             3057..3068
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    3218..3265
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     CDS             complement(3381..4196)
                     /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     4104..4123
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4188..4207
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(4554..5504)
                     /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      5552..5774
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 ggtctgctat gtggtgctat ctgacttttt gctgttcagc agttcctgcc ctctgatttt
       61 ccagtctgac cacttcggat tatcccgtga caggtcattc agactggcta atgcacccag
      121 taaggcagcg gtatcatcaa caggcttacc cgtcttactg tccctagtgc ttggattctc
      181 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga
      241 ggtcattact ggatctatca acaggagtcc aagcgagctc gtaaacttgg tctgacagct
      301 ctagctccgg caaaaaaacg ggcaaggtgt caccaccctg ccctttttct ttaaaaccga
      361 aaagattact tcgcgtttgc cacctgacgt ctaagaactc ggataccctt actctgttga
      421 aaacgaatag ataggtttta tcatttgtac agttcatcca taccatgcgt gatgcccgct
      481 gcggttacga actccagcag aaccatatga tcgcgtttct cgttcggatc tttagacaga
      541 acgctttgcg tgctcagata gtgattgtct ggcagcagaa caggaccatc accgattgga
      601 gtgttttgct ggtagtgatc agccagctgc acgctgccat cctccacgtt gtggcgaatt
      661 ttaaaattcg ctttaatgcc atttttttgt ttatcggcgg tgatgtaaac attgtggctg
      721 ttaaaattgt attccagctt atggcccagg atattgccgt cttctttaaa gtcaatgcct
      781 ttcagctcaa tgcggtttac cagggtatcg ccttcaaatt tcacttccgc acgcgttttg
      841 tacgtgccgt catccttaaa ggaaatcgtg cgttcctgca catagccttc cggcatggcg
      901 gacttgaaga agtcatgctg cttcatatgg tccggataac gagcaaagca ctgaacacca
      961 taagtcagcg tcgttaccag agtcggccaa ggtaccggca gtttaccagt agtacagatg
     1021 aacttcagcg tcagtttacc attagttgcg tcaccttcac cctcgccacg cacggaaaac
     1081 ttatgaccgt tgacatcacc atccagttcc accagaatag ggacgacacc agtgaacagc
     1141 tcttcgcctt tacgcatcta gtatttctcc tctttttaaa caaaattatt tgtagagggg
     1201 tgtttcgtcc tttcggactc atcagtcaag gtacgcacct tgagacaccc tatgccaaca
     1261 caattaacat ctcaatcaag gtaaatgctt tttgcttttt ttgcttgaca attaatcatc
     1321 cggctcgtaa tgtttgtgga tcggccggct tgtcgacgac ggcggtctcc gtcgtcagga
     1381 tcatccgggc agctgtcacc ggatgtgctt tccggtctga tgagtccgtg aggacgaaac
     1441 agcctctaca aataattttg tttaaaaaga ggagaaatac tagatgagcg agctgattaa
     1501 ggagaacatg cacatgaagc tgtacatgga gggcaccgtg gacaaccatc acttcaagtg
     1561 cacatccgag ggcgaaggca agccctacga gggcacccag accatgagaa tcaaggtggt
     1621 cgagggcggc cctctcccct tcgccttcga catcctggct actagcttcc tctacggcag
     1681 caagaccttc atcaaccaca cccagggcat ccccgacttc ttcaagcagt ccttccctga
     1741 gggcttcaca tgggagagag tcaccacata cgaagacggg ggcgtgctga ccgctaccca
     1801 ggacaccagc ctccaggacg gctgcctcat ctacaacgtc aagatcagag gggtgaactt
     1861 cacatccaac ggccctgtga tgcagaagaa aacactcggc tgggaggcct tcaccgagac
     1921 gctgtacccc gctgacggcg gcctgtaagg cagaaacgac atggccctga agctcgtggg
     1981 cgggagccat ctgatcgcaa acatcaagac cacatataga tccaagaaac ccgctaagaa
     2041 cctcaagatg cctggcgtct actatgtgga ctacagactg gaaagaatca aggaggccaa
     2101 caacgagacc tacgtcgagc agcacgaggt ggcagtggcc agatactgcg acctccctag
     2161 caaactgggg cactaaaggc aactgaaacg attcggatcc tgtattacta ttcttatcac
     2221 actggctcac cttcgggtgg gcctttctgc gtttatatac tagagagaga atataaaaag
     2281 ccagattatt aatccggctt ttttattatt tactttatct gagaatagtc aatcttcgga
     2341 aatcccaggt gttatcaacg atgtcctaat ttcgacggaa ggtcacaata acgggctacg
     2401 gctacttcat gttgttcaac gtaggtttct ttatcagctt ctttaatacg ttctaagcga
     2461 cgatcaacat agtatactcc aggcatttta aggtttttag cgggtttttt ggaacgatag
     2521 gttgttttca ggttgcagat taaatgaccg cctccaacta atttcagtgc catgtctgcg
     2581 cgtccttcta agccaccgtc cgcaggatat aaggtttctg ttgatgcttc ccaaccaaga
     2641 gttttttttt gcattacggg tccgttactc gggaaattta cgccacggat tttaacattg
     2701 taaatcagac agccatcttg taaactcgta tcttgagttg ctgtaagtac accgccatct
     2761 tcataagttg ttacacgttc ccaggtaaag ccttcaggaa aggactgttt aaaaaagtca
     2821 ggaatgcctt gggtgtgatt gataaaagtt ttagacccgt acataaagct cgttgcgaga
     2881 atgtcaaacg caaagggcag gggtccacct tcaactgctt tgatgcgcat tgtttgtgtg
     2941 ccttcatatg gtttaccttc accttcggat gtgcatttga aatgatgatt gttgacagta
     3001 ccttccatat ataatttcat gtgcatattt tctttaatta attctgacat ctagtatttc
     3061 tcctcttttt aaacaaaatt atttgtagag gtggtttcgc tctgccttga gctcatcagg
     3121 tgatcgagtt tcagatcacg accactgcga gtttttattt cgtttatttc aattaaggta
     3181 actaaaaaac tcctttactc aggtctgccg taaggggggt ttgtaccgta caccactgag
     3241 accgcggtgg ttgaccagac aaaccacgaa ttattgacca cttccgagta gaatcgtgct
     3301 tcagtaagag ctcactcaaa ggcggtaatc agataaaaaa aatccttagc tttcgctaag
     3361 gatgatttct gctagtatta ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
     3421 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     3481 aactcaccga ggcagttcca aagaatggca aggtcctggt aacggtctgc gattccgacc
     3541 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     3601 aaatcaccat gagtgacgac tgaatccggt gagaatggca agagcttgtg catttctttc
     3661 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     3721 ccgttattca tgcgtgattg cgcctgagca agacgaaata cacgatcgct gttaaaagga
     3781 caattacaaa caggaatcga atgtaaccgg cgcaggaaca cggccagcgc atcaacaata
     3841 ttttcacctg aatcaggata ttcttctaat acctggaagg ctgttttccc aggaatcgcg
     3901 gtggtgagta accacgcatc atcaggagta cggataaaat gcttgatggt cgggagaggc
     3961 ataaactccg tcagccagtt gagacggacc atctcatctg taacatcatt ggcaacgcta
     4021 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa gcgatagatt
     4081 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcgtcc
     4141 atgttggagt ttaagcgcgg acgggagcaa gacgtttccc gttgaatatg gctcataaca
     4201 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta
     4261 tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttgttg aataaatcga
     4321 acttttgctg agttgaagaa tcagctctag tagttacatt gtcgatctgt tcatggtgaa
     4381 cagctttgaa tgcaccaaaa actcgtaaaa gctctgatgt atctatcttt tttacaccgt
     4441 tttcatctgt gcatatggac agttttccct ttgatatgta acggtgaaca gttgttctac
     4501 ttttgtttgt tagtcttgat gcttcactga tagatacaag agccataaga acctcagatc
     4561 cttccgtatt tagccagtat gttctctagt gtggttcgtt gtttttgcgt gagccatgag
     4621 aacgaaccat tgagatcata cttactttgc atgtcactca aaaattttgc ctcaaaactg
     4681 gtgagctgaa tttttgcagt taaagcatcg tgtagtgttt ttcttagtcc gttatgtagg
     4741 taggaatctg atgtaatggt tgttggtatt ttgtcaccat tcatttttat ctggttgttc
     4801 tcaagttcgg ttacgagatc catttgtcta tctagttcaa cttggaaaat caacgtatca
     4861 gtcgggcggc ctcgcttatc aaccaccaat ttcatattgc tgtaagtgtt taaatcttta
     4921 cttattggtt tcaaaaccca ttggttaagc cttttaaact catggtagtt attttcaagc
     4981 attaacatga acttaaattc atcaaggcta atctctatat ttgccttgtg agttttcttt
     5041 tgtgttagtt cttttaataa ccactcataa atcctcatag agtatttgtt ttcaaaagac
     5101 ttaacatgtt ccagattata ttttatgaat ttttttaact ggaaaagata aggcaatatc
     5161 tcttcactaa aaactaattc taatttttcg cttgagaact tggcatagtt tgtccactgg
     5221 aaaatctcaa agcctttaac caaaggattc ctgatttcca cagttctcgt catcagctct
     5281 ctggttgctt tagctaatac accataagca ttttccctac tgatgttcat catctgagcg
     5341 tattggttat aagtgaacga taccgtccgt tctttccttg tagggttttc aatcgtgggg
     5401 ttgagtagtg ccacacagca taaaattagc ttggtttcat gctccgttaa gtcatagcga
     5461 ctaatcgcta gttcatttgc tttgaaaaca actaattcag acatacatct caattggtct
     5521 aggtgatttt aatcactata ccaattgaga tgggctagtc aatgataatt acatgtcctt
     5581 ttcctttgag ttgtgggtat ctgtaaattc tgctagacct ttgctggaaa acttgtaaat
     5641 tctgctagac cctctgtaaa ttccgctaga cctttgtgtg ttttttttgt ttatattcaa
     5701 gtggttataa tttatagaat aaagaaagaa taaaaaaaga taaaaagaat agatcccagc
     5761 cctgtgtata actcactact ttagtcagtt ccgcagtatt acaaaaggat gtcgcaaacg
     5821 ctgtttgctc ctctacaaaa cagaccttaa aaccctaaag gcttaagtag caccctcgca
     5881 agctcgggca aatcgctgaa tattcctttt gtctccgacc atcaggcacc tgagtcgctg
     5941 tctttttcgt gacattcagt tcgctgcgct cacggctctg gcagtgaatg ggggtaaatg
     6001 gcactacagg cgccttttat ggattcatgc aaggaaacta cccataatac aagaaaagcc
     6061 cgtcacgggc ttctcagggc gttttatggc g
//
