LOCUS       sequence-250098-        6139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2-input history dependent program B then A; sfGFP-GFP221-GFP226.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6139)
  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 6139)
  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: 2SP1
FEATURES             Location/Qualifiers
     source          1..6139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(722..749)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(803..1516)
                     /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             1523..1534
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_RNA        complement(1558..1607)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     protein_bind    1699..1748
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     RBS             1825..1836
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(1843..1845,1846..1848,1849..2559)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      2731..2825
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3317..3539
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             3587..4537
                     /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(4884..4903)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4895..5710
                     /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(4968..4987)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     protein_bind    5893..5940
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     RBS             6023..6034
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(6041..6043,6044..6046,6047..6139,1..618)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
ORIGIN
        1 ggtgagggtg atgctacata cgggaaattg actcttaaat ttatatgtac aacaggtaaa
       61 ctacctgttc catggcctac acttgtcact acgcttactt acggggtcca gtgcttctct
      121 agatatcctg atcacatgaa gcaacatgac tttttcaaga gcgccatgcc cgagggatat
      181 gtgcaggaga ggaccatatt tttcaaggat gatgggaact acaaaacgcg cgcggaggtc
      241 aaattcgaag gcgacacact cgtaaacaga atcgagttaa aaggaatcga cttcaaagaa
      301 gatgggaaca tccttgggca caaattggag tataactata actctcataa tgtttatata
      361 atggcagata aacaaaaaaa cggaataaag gttaacttca aaatcagaca caacatcgaa
      421 gacggtagcg tacagctggc cgatcactat cagcagaaca ccccgatcgg tgatggtccg
      481 gtactgttgc cggataacca ctacctgtcc actcagtctg ccctctccaa agacccgaac
      541 gaaaaaagag atcacatggt tctgctggag ttcgtaaccg ctgcgggcat caccctgggc
      601 atggacgaac tgtacaaata acacctggga tttccgaaga ttgactattc tcagataaag
      661 taaataataa aaaagccgga ttaataatct ggctttttat attctctctc tagtatataa
      721 acgcagaaag gcccacccga aggtgagcca gtgtgataag aatagtaata caggatccga
      781 atcgtttcag ttgcctttat catttgtaca gttcatccat accatgcgtg atgcccgctg
      841 cggttacgaa ctccagcaga accatatgat cgcgtttctc gttcggatct ttagacagaa
      901 cgctttgcgt gctcagatag tgattgtctg gcagcagaac aggaccatca ccgattggag
      961 tgttttgctg gtagtgatca gccagctgca cgctgccatc ctccacgttg tggcgaattt
     1021 taaaattcgc tttaatgcca tttttttgtt tatcggcggt gatgtaaaca ttgtggctgt
     1081 taaaattgta ttccagctta tggcccagga tattgccgtc ttctttaaag tcaatgcctt
     1141 tcagctcaat gcggtttacc agggtatcgc cttcaaattt cacttccgca cgcgttttgt
     1201 acgtgccgtc atccttaaag gaaatcgtgc gttcctgcac atagccttcc ggcatggcgg
     1261 acttgaagaa gtcatgctgc ttcatatggt ccggataacg agcaaagcac tgaacaccat
     1321 aagtcagcgt cgttaccaga gtcggccaag gtaccggcag tttaccagta gtacagatga
     1381 acttcagcgt cagtttacca ttagttgcgt caccttcacc ctcgccacgc acggaaaact
     1441 tatgaccgtt gacatcacca tccagttcca ccagaatagg gacgacacca gtgaacagct
     1501 cttcgccttt acgcatctag tatttctcct ctttttaaac aaaattattt gtagaggctg
     1561 tttcgtcctc acggactcat cagaccggaa agcacatccg gtgacagctg caaaaaaagc
     1621 aaaaagcatt taccttgatt gagatgttaa ttgtgttggc attccacaaa cattacgagc
     1681 cggatgatta attgtcaagc ccggatgatc ctgacgacgg agaccgccgt cgtcgacaag
     1741 ccggccgaag ggtgtctcaa ggtgcgtacc ttgactgatg agtccgaaag gacgaaacac
     1801 ccctctacaa ataattttgt ttaaaaagag gagaaatact agatggtgag caaaggtgag
     1861 gaacttttta caggagtagt acctattctt gtagagttag atggtgatgt gaatggccat
     1921 aaatttagtg tttctggtga aggagagggc gatgcgactt atggcaagtt gacccttaaa
     1981 tttatttgta ctactggtaa acttcctgtt ccttggccta ctctcgtgac aactcttact
     2041 tacggagtcc aatgcttctc tagatacccg gaccacatga agcagcacga tttcttcaaa
     2101 agcgccatgc ctgagggata cgtccaagaa aggacgatct tcttcaagga cgacgggaac
     2161 tacaagacac gggcggaggt gaaatttgag ggggacacgc tcgtgaacag gattgagttg
     2221 aaggggattg acttcaagga ggacggcaat atactcggac acaaattaga gtacaactac
     2281 aactcgcaca acgtctacat tatggccgat aagcagaaga acggcattaa ggttaacttc
     2341 aagatcaggc acaatatcga ggatggcagc gttcagcttg cagatcatta ccaacagaac
     2401 acaccgatcg gcgacggccc ggtactgttg ccggataacc actacctgtc gacgcagtct
     2461 gccctatcga aagaccctaa cgaaaaaaga gaccatatgg tgctattaga gtttgtgact
     2521 gcagcaggta ttacacttgg tatggacgaa ctttataaat aaaacctatc tattcgtttt
     2581 caacagagta agggtatccg agttcttaga cgtcaggtgg caaacgcgaa gtaatctttt
     2641 cggttttaaa gaaaaagggc agggtggtga caccttgccc gtttttttgc cggagctaga
     2701 gctgtcagac caagtttacg agctcgcttg gactcctgtt gatagatcca gtaatgacct
     2761 cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg
     2821 agaatccaag cactagggac agtaagacgg gtaagcctgt tgatgatacc gctgccttac
     2881 tgggtgcatt agccagtctg aatgacctgt cacgggataa tccgaagtgg tcagactgga
     2941 aaatcagagg gcaggaactg ctgaacagca aaaagtcaga tagcaccaca tagcagaccc
     3001 gccataaaac gccctgagaa gcccgtgacg ggcttttctt gtattatggg tagtttcctt
     3061 gcatgaatcc ataaaaggcg cctgtagtgc catttacccc cattcactgc cagagccgtg
     3121 agcgcagcga actgaatgtc acgaaaaaga cagcgactca ggtgcctgat ggtcggagac
     3181 aaaaggaata ttcagcgatt tgcccgagct tgcgagggtg ctacttaagc ctttagggtt
     3241 ttaaggtctg ttttgtagag gagcaaacag cgtttgcgac atccttttgt aatactgcgg
     3301 aactgactaa agtagtgagt tatacacagg gctgggatct attcttttta tcttttttta
     3361 ttctttcttt attctataaa ttataaccac ttgaatataa acaaaaaaaa cacacaaagg
     3421 tctagcggaa tttacagagg gtctagcaga atttacaagt tttccagcaa aggtctagca
     3481 gaatttacag atacccacaa ctcaaaggaa aaggacatgt aattatcatt gactagccca
     3541 tctcaattgg tatagtgatt aaaatcacct agaccaattg agatgtatgt ctgaattagt
     3601 tgttttcaaa gcaaatgaac tagcgattag tcgctatgac ttaacggagc atgaaaccaa
     3661 gctaatttta tgctgtgtgg cactactcaa ccccacgatt gaaaacccta caaggaaaga
     3721 acggacggta tcgttcactt ataaccaata cgctcagatg atgaacatca gtagggaaaa
     3781 tgcttatggt gtattagcta aagcaaccag agagctgatg acgagaactg tggaaatcag
     3841 gaatcctttg gttaaaggct ttgagatttt ccagtggaca aactatgcca agttctcaag
     3901 cgaaaaatta gaattagttt ttagtgaaga gatattgcct tatcttttcc agttaaaaaa
     3961 attcataaaa tataatctgg aacatgttaa gtcttttgaa aacaaatact ctatgaggat
     4021 ttatgagtgg ttattaaaag aactaacaca aaagaaaact cacaaggcaa atatagagat
     4081 tagccttgat gaatttaagt tcatgttaat gcttgaaaat aactaccatg agtttaaaag
     4141 gcttaaccaa tgggttttga aaccaataag taaagattta aacacttaca gcaatatgaa
     4201 attggtggtt gataagcgag gccgcccgac tgatacgttg attttccaag ttgaactaga
     4261 tagacaaatg gatctcgtaa ccgaacttga gaacaaccag ataaaaatga atggtgacaa
     4321 aataccaaca accattacat cagattccta cctacataac ggactaagaa aaacactaca
     4381 cgatgcttta actgcaaaaa ttcagctcac cagttttgag gcaaaatttt tgagtgacat
     4441 gcaaagtaag tatgatctca atggttcgtt ctcatggctc acgcaaaaac aacgaaccac
     4501 actagagaac atactggcta aatacggaag gatctgaggt tcttatggct cttgtatcta
     4561 tcagtgaagc atcaagacta acaaacaaaa gtagaacaac tgttcaccgt tacatatcaa
     4621 agggaaaact gtccatatgc acagatgaaa acggtgtaaa aaagatagat acatcagagc
     4681 ttttacgagt ttttggtgca ttcaaagctg ttcaccatga acagatcgac aatgtaacta
     4741 ctagagctga tccttcaact cagcaaaagt tcgatttatt caacaaagcc acgttgtgtc
     4801 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     4861 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     4921 ttgctcccgt ccgcgcttaa actccaacat ggacgctgat ttatatgggt ataaatgggc
     4981 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     5041 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     5101 ggtccgtctc aactggctga cggagtttat gcctctcccg accatcaagc attttatccg
     5161 tactcctgat gatgcgtggt tactcaccac cgcgattcct gggaaaacag ccttccaggt
     5221 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggccg tgttcctgcg
     5281 ccggttacat tcgattcctg tttgtaattg tccttttaac agcgatcgtg tatttcgtct
     5341 tgctcaggcg caatcacgca tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     5401 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cacaagctct tgccattctc
     5461 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     5521 gaaattaata ggttgtattg atgttggacg ggtcggaatc gcagaccgtt accaggacct
     5581 tgccattctt tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     5641 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     5701 gtttttctaa taatactagc agaaatcatc cttagcgaaa gctaaggatt ttttttatct
     5761 gattaccgcc tttgagtgag ctcttactga agcacgattc tactcggaag tggtcaataa
     5821 tgcgagtttt tatttcgttt atttcaatta aggtaactaa aaaactcctt tcccttacgg
     5881 cagacctgag tgggtttgta ccgtacacca ctgagaccgc ggtggttgac cagacaaacc
     5941 acgaagtggt cgtgatctga aactcgatca cctgatgagc tcaaggcaga gcgaaaccac
     6001 ctctacaaat aattttgttt aaaaagagga gaaatactag atggttagta aaggcgaaga
     6061 actatttaca ggtgtagtac caatactagt agagttggat ggggatgtaa atggtcataa
     6121 atttagtgtt tcaggtgaa
//
