LOCUS       sequence-251838-        5305 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2-input history dependent program A then B;0-mKate2-sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5305)
  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 5305)
  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: 2SP7
FEATURES             Location/Qualifiers
     source          1..5305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      815..909
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1401..1623
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             1671..2621
                     /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(2968..2987)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2979..3794
                     /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(3052..3071)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     protein_bind    3910..3957
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     RBS             4107..4118
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      complement(4924..4951)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    4999..5048
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     RBS             5214..5225
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(5232..5305,1..640)
                     /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"
ORIGIN
        1 taagttttcc gtgcgtggcg agggtgaagg tgacgcaact aatggtaaac tgacgctgaa
       61 gttcatctgt actactggta aactgccggt accttggccg actctggtaa cgacgctgac
      121 ttatggtgtt cagtgctttg ctcgttatcc ggaccatatg aagcagcatg acttcttcaa
      181 gtccgccatg ccggaaggct atgtgcagga acgcacgatt tcctttaagg atgacggcac
      241 gtacaaaacg cgtgcggaag tgaaatttga aggcgatacc ctggtaaacc gcattgagct
      301 gaaaggcatt gactttaaag aagacggcaa tatcctgggc cataagctgg aatacaattt
      361 taacagccac aatgtttaca tcaccgccga taaacaaaaa aatggcatta aagcgaattt
      421 taaaattcgc cacaacgtgg aggatggcag cgtgcagctg gctgatcact accagcaaaa
      481 cactccaatc ggtgatggtc ctgttctgct gccagacaat cactatctga gcacgcaaag
      541 cgttctgtct aaagatccga acgagaaacg cgatcatatg gttctgctgg agttcgtaac
      601 cgcagcgggc atcacgcatg gtatggatga actgtacaaa tgataaaacc tatctattcg
      661 ttttcaacag agtaagggta tccgagttct tagacgtcag gtggcaaacg cgaagtaatc
      721 ttttcggttt taaagaaaaa gggcagggtg gtgacacctt gcccgttttt ttgccggagc
      781 tagagctgtc agaccaagtt tacgagctcg cttggactcc tgttgataga tccagtaatg
      841 acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt
      901 ggtgagaatc caagcactag ggacagtaag acgggtaagc ctgttgatga taccgctgcc
      961 ttactgggtg cattagccag tctgaatgac ctgtcacggg ataatccgaa gtggtcagac
     1021 tggaaaatca gagggcagga actgctgaac agcaaaaagt cagatagcac cacatagcag
     1081 acccgccata aaacgccctg agaagcccgt gacgggcttt tcttgtatta tgggtagttt
     1141 ccttgcatga atccataaaa ggcgcctgta gtgccattta cccccattca ctgccagagc
     1201 cgtgagcgca gcgaactgaa tgtcacgaaa aagacagcga ctcaggtgcc tgatggtcgg
     1261 agacaaaagg aatattcagc gatttgcccg agcttgcgag ggtgctactt aagcctttag
     1321 ggttttaagg tctgttttgt agaggagcaa acagcgtttg cgacatcctt ttgtaatact
     1381 gcggaactga ctaaagtagt gagttataca cagggctggg atctattctt tttatctttt
     1441 tttattcttt ctttattcta taaattataa ccacttgaat ataaacaaaa aaaacacaca
     1501 aaggtctagc ggaatttaca gagggtctag cagaatttac aagttttcca gcaaaggtct
     1561 agcagaattt acagataccc acaactcaaa ggaaaaggac atgtaattat cattgactag
     1621 cccatctcaa ttggtatagt gattaaaatc acctagacca attgagatgt atgtctgaat
     1681 tagttgtttt caaagcaaat gaactagcga ttagtcgcta tgacttaacg gagcatgaaa
     1741 ccaagctaat tttatgctgt gtggcactac tcaaccccac gattgaaaac cctacaagga
     1801 aagaacggac ggtatcgttc acttataacc aatacgctca gatgatgaac atcagtaggg
     1861 aaaatgctta tggtgtatta gctaaagcaa ccagagagct gatgacgaga actgtggaaa
     1921 tcaggaatcc tttggttaaa ggctttgaga ttttccagtg gacaaactat gccaagttct
     1981 caagcgaaaa attagaatta gtttttagtg aagagatatt gccttatctt ttccagttaa
     2041 aaaaattcat aaaatataat ctggaacatg ttaagtcttt tgaaaacaaa tactctatga
     2101 ggatttatga gtggttatta aaagaactaa cacaaaagaa aactcacaag gcaaatatag
     2161 agattagcct tgatgaattt aagttcatgt taatgcttga aaataactac catgagttta
     2221 aaaggcttaa ccaatgggtt ttgaaaccaa taagtaaaga tttaaacact tacagcaata
     2281 tgaaattggt ggttgataag cgaggccgcc cgactgatac gttgattttc caagttgaac
     2341 tagatagaca aatggatctc gtaaccgaac ttgagaacaa ccagataaaa atgaatggtg
     2401 acaaaatacc aacaaccatt acatcagatt cctacctaca taacggacta agaaaaacac
     2461 tacacgatgc tttaactgca aaaattcagc tcaccagttt tgaggcaaaa tttttgagtg
     2521 acatgcaaag taagtatgat ctcaatggtt cgttctcatg gctcacgcaa aaacaacgaa
     2581 ccacactaga gaacatactg gctaaatacg gaaggatctg aggttcttat ggctcttgta
     2641 tctatcagtg aagcatcaag actaacaaac aaaagtagaa caactgttca ccgttacata
     2701 tcaaagggaa aactgtccat atgcacagat gaaaacggtg taaaaaagat agatacatca
     2761 gagcttttac gagtttttgg tgcattcaaa gctgttcacc atgaacagat cgacaatgta
     2821 actactagag ctgatccttc aactcagcaa aagttcgatt tattcaacaa agccacgttg
     2881 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
     2941 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     3001 cgtcttgctc ccgtccgcgc ttaaactcca acatggacgc tgatttatat gggtataaat
     3061 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg
     3121 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     3181 agatggtccg tctcaactgg ctgacggagt ttatgcctct cccgaccatc aagcatttta
     3241 tccgtactcc tgatgatgcg tggttactca ccaccgcgat tcctgggaaa acagccttcc
     3301 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gccgtgttcc
     3361 tgcgccggtt acattcgatt cctgtttgta attgtccttt taacagcgat cgtgtatttc
     3421 gtcttgctca ggcgcaatca cgcatgaata acggtttggt tgatgcgagt gattttgatg
     3481 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcacaag ctcttgccat
     3541 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     3601 aggggaaatt aataggttgt attgatgttg gacgggtcgg aatcgcagac cgttaccagg
     3661 accttgccat tctttggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     3721 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     3781 atgagttttt ctaataatac tagcagaaat catccttagc gaaagctaag gatttttttt
     3841 atctgattac cgcctttgag tgagctctta ctgaagcacg attctactcg gaagtggtca
     3901 ataattcgtg gtttgtctgg tcaaccaccg cggtctcagt ggtgtacggt acaaaccccc
     3961 cttacggcag acctgagtaa aggagttttt tagttacctt aattgaaata aacgaaataa
     4021 aaactcgcag tggtcgtgat ctgaaactcg atcacctgat gagctcaagg cagagcgaaa
     4081 ccacctctac aaataatttt gtttaaaaag aggagaaata ctagatgtca gaattaatta
     4141 aagaaaatat gcacatgaaa ttatatatgg aaggtactgt caacaatcat catttcaaat
     4201 gcacatccga aggtgaaggt aaaccatatg aaggcacaca aacaatgcgc atcaaagcag
     4261 ttgaaggtgg acccctgccc tttgcgtttg acattctcgc aacgagcttt atgtacgggt
     4321 ctaaaacttt tatcaatcac acccaaggca ttcctgactt ttttaaacag tcctttcctg
     4381 aaggctttac ctgggaacgt gtaacaactt atgaagatgg cggtgtactt acagcaactc
     4441 aagatacgag tttacaagat ggctgtctga tttacaatgt taaaatccgt ggcgtaaatt
     4501 tcccgagtaa cggacccgta atgcaaaaaa aaactcttgg ttgggaagca tcaacagaaa
     4561 ccttatatcc tgcggacggt ggcttagaag gacgcgcaga catggcactg aaattagttg
     4621 gaggcggtca tttaatctgc aacctgaaaa caacctatcg ttccaaaaaa cccgctaaaa
     4681 accttaaaat gcctggagta tactatgttg atcgtcgctt agaacgtatt aaagaagctg
     4741 ataaagaaac ctacgttgaa caacatgaag tagccgtagc ccgttattgt gaccttccgt
     4801 cgaaattagg acatcgttga taacacctgg gatttccgaa gattgactat tctcagataa
     4861 agtaaataat aaaaaagccg gattaataat ctggcttttt atattctctc tctagtatat
     4921 aaacgcagaa aggcccaccc gaaggtgagc cagtgtgata agaatagtaa tacaggatcc
     4981 gaatcgtttc agttgcctgc ccggatgatc ctgacgacgg agaccgccgt cgtcgacaag
     5041 ccggccgatc cacaaacatt acgagccgga tgattaattg tcaagcaaaa aaagcaaaaa
     5101 gcatttacct tgattgagat gttaattgtg ttggcatagg gtgtctcaag gtgcgtacct
     5161 tgactgatga gtccgaaagg acgaaacacc cctctacaaa taattttgtt taaaaagagg
     5221 agaaatacta gatgcgtaaa ggcgaagagc tgttcactgg tgtcgtccct attctggtgg
     5281 aactggatgg tgatgtcaac ggtca
//
