LOCUS       sequence-377947-        3217 bp    DNA     circular SYN 15-APR-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 3217)
  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 3217)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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..3217
                     /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             334..1128
                     /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(388..407)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     998..1017
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      1149..1220
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1236..1263
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1291..1308)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1425..1970)
                     /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(1505..1524)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2084..2178)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2199..2858)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2773..2792
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2859..2961)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3122..3141
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ttatcaaaaa gagtattgca ttaaagtcta acctatagga atcttacagc 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 cgaagttata tcactctacg gccagctgca aacatgattg aacaagatgg attgcacgca
      361 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
      421 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
      481 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
      541 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
      601 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
      661 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
      721 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
      781 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
      841 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
      901 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
      961 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     1021 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     1081 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgata atagggatct
     1141 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     1201 gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct
     1261 gcgtttatac ctagggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt
     1321 tcgactgcgg cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag
     1381 gaagatactt aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc
     1441 ccccctgaca agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga
     1501 ctataaagat accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc
     1561 tttcggttta ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac
     1621 actcagttcc gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca
     1681 gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc
     1741 aaaagcacca ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat
     1801 gcgccggtta aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt
     1861 tacctcggtt caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg
     1921 ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc
     1981 ttattaatca gataaaatat ttctagattt cagtgcaatt tatctcttca aatgtagcac
     2041 ctgaagtcag ccccatacga tataagttgt tactagtgct tggattctca ccaataaaaa
     2101 acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg
     2161 gatctatcaa caggagtcca agcgagctcg atatcaaatt acgccccgcc ctgccactca
     2221 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca
     2281 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
     2341 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
     2401 aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa
     2461 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
     2521 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
     2581 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacga
     2641 aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
     2701 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
     2761 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
     2821 atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa
     2881 aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg
     2941 gaacctctta cgtgccgatc aacgtctcat tttcgccaga tatcgacgtc ggtgcctaat
     3001 gagtgagcta acttacatta attgcgttgc gcattcacca ccctgaattg actctcttcc
     3061 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgga tcttccccat
     3121 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     3181 cgatgccgaa gttcctatta tcacgaggcc ctttgag
//
