LOCUS       sequence-377954-        3466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-inducible activation of cat-T172A for chloramphenicol 
            resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3466)
  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 3466)
  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-cat-T172A-P-R
FEATURES             Location/Qualifiers
     source          1..3466
                     /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..989
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFAMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(396..415)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1049..1120
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1136..1163
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1191..1208)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1325..1870)
                     /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(1405..1424)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1984..2078)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2109..2903)
                     /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(2220..2239)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2830..2849
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3088..3165)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3371..3390
                     /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 cgaagttatt ttaagaagga gatatacata tggagaaaaa aatcactgga tataccaccg
      361 ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat
      421 gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa
      481 ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc
      541 cggaatttcg tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt
      601 gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg
      661 acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc
      721 tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc aatccctggg
      781 tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt
      841 tcgccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg
      901 ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt
      961 actgcgatga gtggcagggc ggggcgtaat aatagggatc tccaggcatc aaataaggat
     1021 ccaaactcga gtaaggatct ccaggcatca aataaaacga aaggctcagt cgaaagactg
     1081 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc
     1141 accttcgggt gggcctttct gcgtttatac ctagggatat attccgcttc ctcgctcact
     1201 gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga acggggcgga
     1261 gatttcctgg aagatgccag gaagatactt aacagggaag tgagagggcc gcggcaaagc
     1321 cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt
     1381 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctggc ggctccctcg
     1441 tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat tccgctgtta tggccgcgtt
     1501 tgtctcattc cacgcctgac actcagttcc gggtaggcag ttcgctccaa gctggactgt
     1561 atgcacgaac cccccgttca gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1621 tccaacccgg aaagacatgc aaaagcacca ctggcagcag ccactggtaa ttgatttaga
     1681 ggagttagtc ttgaagtcat gcgccggtta aggctaaact gaaaggacaa gttttggtga
     1741 ctgcgctcct ccaagccagt tacctcggtt caaagagttg gtagctcaga gaaccttcga
     1801 aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa
     1861 acgatctcaa gaagatcatc ttattaatca gataaaatat ttctagattt cagtgcaatt
     1921 tatctcttca aatgtagcac ctgaagtcag ccccatacga tataagttgt tactagtgct
     1981 tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg
     2041 gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctct cgaaccccag
     2101 agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga
     2161 gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca
     2221 atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag
     2281 tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca
     2341 tgggtcacga cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg
     2401 gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc
     2461 atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc
     2521 ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga
     2581 gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt
     2641 cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac
     2701 gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc ggtcttgaca
     2761 aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt
     2821 gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg
     2881 tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagatcat
     2941 gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac tttgcagggc
     3001 ttcccaacct taccagaggg cgccccagct ggcaattccg acgtcggtgc ctaatgagtg
     3061 agctaactta cattaattgc gttgcgcatt caccaccctg aattgactct cttccgggcg
     3121 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     3181 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     3241 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     3301 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     3361 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     3421 tgccggccac gatgccgaag ttcctattat cacgaggccc tttgag
//
