LOCUS       sequence-377956-        3427 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-inducible activation of cat
            amphenicol resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3427)
  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 3427)
  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-P-R
FEATURES             Location/Qualifiers
     source          1..3427
                     /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
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(396..415)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1010..1081
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1097..1124
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1152..1169)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1286..1831)
                     /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(1366..1385)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1945..2039)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2070..2864)
                     /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(2181..2200)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2791..2810
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3049..3126)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3332..3351
                     /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 tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg
      901 ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt
      961 actgcgatga gtggcagggc ggggcgtaat aatagggatc tccaggcatc aaataaaacg
     1021 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     1081 ctactagagt cacactggct caccttcggg tgggcctttc tgcgtttata cctagggata
     1141 tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa
     1201 atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa
     1261 gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg
     1321 aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt
     1381 ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca
     1441 ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca
     1501 gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta
     1561 tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca
     1621 gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac
     1681 tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt
     1741 ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca
     1801 agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata
     1861 tttctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg
     1921 atataagttg ttactagtgc ttggattctc accaataaaa aacgcccggc ggcaaccgag
     1981 cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc
     2041 aagcgagctc tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag
     2101 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat
     2161 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc
     2221 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata
     2281 ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgcgcgcc
     2341 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc
     2401 tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg
     2461 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg
     2521 atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg
     2581 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga
     2641 acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca
     2701 ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg
     2761 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc
     2821 caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat
     2881 cctgtctctt gatcagatca tgatcccctg cgccatcaga tccttggcgg caagaaagcc
     2941 atccagttta ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc
     3001 gacgtcggtg cctaatgagt gagctaactt acattaattg cgttgcgcat tcaccaccct
     3061 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     3121 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     3181 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     3241 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     3301 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     3361 ccgcacctgt ggcgccggtg atgccggcca cgatgccgaa gttcctatta tcacgaggcc
     3421 ctttgag
//
