LOCUS       sequence-377953-        3410 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-inducible activation of knt for kanamycin resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3410)
  AUTHORS   Sheets MB, Tague N, Dunlop MJ
  TITLE     An optogenetic toolkit for light-inducible antibiotic resistance.
  JOURNAL   Nat Commun. 2023 Feb 23;14(1):1034. doi: 10.1038/s41467-023-36670-2.
  PUBMED    36823420
REFERENCE   2  (bases 1 to 3410)
  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: "2023-02-23"; volume: "14(1):1034. doi"; pages: " 
            10.1038/s41467-023-36670-2"
COMMENT     Sequence Label: p15A-OptoCre-knt-P*-R
FEATURES             Location/Qualifiers
     source          1..3410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(72..105)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      153..224
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      240..267
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    complement(276..309)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             330..1124
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(384..403)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     994..1013
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      1145..1216
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1232..1259
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1287..1304)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1421..1966)
                     /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(1501..1520)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2080..2174)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2195..2854)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2769..2788
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2855..2957)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(3221..3254)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     3315..3334
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ttatcaaaaa gagtattgtc ttaaagtcta acctatagga ttcttacagc catcgagagg
       61 gacacggcga aataacttcg tatagcatac attatacgaa gttatccagg catcaaataa
      121 ggatccaaac tcgagtaagg atctccaggc atcaaataaa acgaaaggct cagtcgaaag
      181 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg
      241 gctcaccttc gggtgggcct ttctgcgttt ataccataac ttcgtatagc atacattata
      301 cgaagttatt ttaagaagga gatatacata tgattgaaca agatggattg cacgcaggtt
      361 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
      421 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
      481 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
      541 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
      601 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
      661 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
      721 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
      781 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
      841 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
      901 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
      961 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     1021 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     1081 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgataatag ggatctccag
     1141 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     1201 gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt
     1261 ttatacctag ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga
     1321 ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag
     1381 atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc
     1441 ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat
     1501 aaagatacca ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc
     1561 ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc
     1621 agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc
     1681 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa
     1741 gcaccactgg cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc
     1801 cggttaaggc taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc
     1861 tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt
     1921 ttcgttttca gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat
     1981 taatcagata aaatatttct agatttcagt gcaatttatc tcttcaaatg tagcacctga
     2041 agtcagcccc atacgatata agttgttact agtgcttgga ttctcaccaa taaaaaacgc
     2101 ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc
     2161 tatcaacagg agtccaagcg agctcgatat caaattacgc cccgccctgc cactcatcgc
     2221 agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat
     2281 gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg
     2341 tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac
     2401 tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg
     2461 ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat
     2521 cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt
     2581 aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc atacgaaatt
     2641 ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct
     2701 tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg
     2761 tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat
     2821 caacggtggt atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc
     2881 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
     2941 ctcttacgtg ccgatcaacg tctcattttc gccagatatc gacaattccg gttcgcttgc
     3001 tgtccataaa accgcccagt ctagctatcg ccatgtaagc ccactgcaag ctacctgctt
     3061 tctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatccgggg
     3121 tcagcaccgt ttctgcggac tggctttcta cgtgttccgc ttcctttagc agcccttgcg
     3181 ccctgagtgc ttgcggcagc gtgagcttca aaagcgctct gaagttccta tactttctag
     3241 agaataggaa cttcgaactg caggtcgacg gatccccgga attaattctc atgtttgaca
     3301 ggatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     3361 gtgatgccgg ccacgatgcc gaagttccta ttatcacgag gccctttgag
//
