LOCUS       sequence-100025-        4219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  aTc-inducible SCRIBE_galK(OFF).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4219)
  AUTHORS   Farzadfard F, Lu TK
  TITLE     Synthetic biology. Genomically encoded analog memory with precise in
            vivo DNA writing in living cell populations.
  JOURNAL   Science. 2014 Nov 14;346(6211):1256272. doi: 
            10.1126/science.1256272.
  PUBMED    25395541
REFERENCE   2  (bases 1 to 4219)
  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: "Science."; 
            date: "2014-11-14"; volume: "346(6211):1256272. doi"; pages: " 
            10.1126/science.1256272"
COMMENT     Sequence Label: pFF761
FEATURES             Location/Qualifiers
     source          1..4219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(7..25,26..31,32..48,49..54,55..80)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    7..25
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    32..50
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1312..2097
                     /codon_start=1
                     /gene="bet"
                     /product="single-stranded DNA binding recombinase in the 
                     lambda Red system"
                     /label=Beta
                     /note="mediates pairing of a single-stranded DNA with a 
                     complementary target"
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
     terminator      2115..2201
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2237..2254)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2371..2916)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2451..2470)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3030..3124)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(3154..3945,3946..4014))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3777..3796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4015..4119)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4187..4205
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagtccc tatcagtgat agagattgac atccctatca gtgatagaga tactgagcac
       61 atcagcagga cgcactgacc ttaattaaat gcgcaccctt agcgagaggt ttatcattaa
      121 ggtcaacctc tggatgttgt ttcggcatcc tgcattgaat ctgagttact gtctgttttc
      181 ctgaattcgg aaactgcttt ggtccccagt gagcggcaat cgattcatta ggcatgatct
      241 ttcttgccga gcgcggaaat tagctggaat tcaggaaacc cgttttttct gacgtaaggg
      301 tgcgcaactt tcatgaaatc cgctgaatat ttgaacactt ttagattgag aaatctcggc
      361 ctacctgtca tgaacaattt gcatgacatg tctaaggcga ctcgcatatc tgttgaaaca
      421 cttcggttgt taatctatac agctgatttt cgctatagga tctacactgt agaaaagaaa
      481 ggcccagaga agagaatgag aaccatttac caaccttctc gagaacttaa agccttacaa
      541 ggatgggttc tacgtaacat tttagataaa ctgtcgtcat ctcctttttc tattggattt
      601 gaaaagcacc aatctatttt gaataatgct accccgcata ttggggcaaa ctttatactg
      661 aatattgatt tggaggattt tttcccaagt ttaactgcta acaaagtttt tggagtgttc
      721 cattctcttg gttataatcg actaatatct tcagttttga caaaaatatg ttgttataaa
      781 aatctgctac cacaaggtgc tccatcatca cctaaattag ctaatctaat atgttctaaa
      841 cttgattatc gtattcaggg ttatgcaggt agtcggggct tgatatatac gagatatgcc
      901 gatgacctca ccttatctgc acagtctatg aaaaaggttg ttaaagcacg tgatttttta
      961 ttttctataa tcccaagtga aggattggtt attaactcaa aaaaaacttg tattagtggg
     1021 cctcgtagtc agaggaaagt tacaggttta gttatttcac aagagaaagt tgggataggt
     1081 agagaaaaat ataaagaaat tagagcaaag atacatcata tattttgcgg taagtcttct
     1141 gagatagaac acgttagggg atggttgtca tttattttaa gtgtggattc aaaaagccat
     1201 aggagattaa taacttatat tagcaaatta gaaaaaaaat atggaaagaa ccctttaaat
     1261 aaagcgaaga cctaaggatc cggttgatat tgattcagag gtataaaacg aatgagtact
     1321 gcactcgcaa cgctggctgg gaagctggct gaacgtgtcg gcatggattc tgtcgaccca
     1381 caggaactga tcaccactct tcgccagacg gcatttaaag gtgatgccag cgatgcgcag
     1441 ttcatcgcat tactgatcgt tgccaaccag tacggcctta atccgtggac gaaagaaatt
     1501 tacgcctttc ctgataagca gaatggcatc gttccggtgg tgggcgttga tggctggtcc
     1561 cgcatcatca atgaaaacca gcagtttgat ggcatggact ttgagcagga caatgaatcc
     1621 tgtacatgcc ggatttaccg caaggaccgt aatcatccga tctgcgttac cgaatggatg
     1681 gatgaatgcc gccgcgaacc attcaaaact cgcgaaggca gagaaatcac ggggccgtgg
     1741 cagtcgcatc ccaaacggat gttacgtcat aaagccatga ttcagtgtgc ccgtctggcc
     1801 ttcggatttg ctggtatcta tgacaaggat gaagccgagc gcattgtcga aaatactgca
     1861 tacactgcag aacgtcagcc ggaacgcgac atcactccgg ttaacgatga aaccatgcag
     1921 gagattaaca ctctgctgat cgccctggat aaaacatggg atgacgactt attgccgctc
     1981 tgttcccaga tatttcgccg cgacattcgt gcatcgtcag aactgacaca ggccgaagca
     2041 gtaaaagctc ttggattcct gaaacagaaa gccgcagagc agaaggtggc agcatgaacg
     2101 cgtgctagag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     2161 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccgccc tagacctagg
     2221 ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag
     2281 cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca
     2341 gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca
     2401 tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca
     2461 ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg
     2521 tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt
     2581 aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg
     2641 ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg
     2701 cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc
     2761 taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa
     2821 gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca
     2881 gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata
     2941 aaatatttct agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc
     3001 atacgatata agttgttact agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa
     3061 ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg
     3121 agtccaagcg agctcgtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3181 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3241 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3301 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3361 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3421 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3481 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3541 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3601 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3661 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3721 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3781 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3841 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3901 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3961 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4021 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4081 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4141 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     4201 gaggcccttt cgtcttcac
//
