LOCUS       sequence-377957-        3653 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-inducible activation of tetA for tetracycline resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3653)
  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 3653)
  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-tetA-Ptet-Rtet
FEATURES             Location/Qualifiers
     source          1..3653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(122..1312)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     941..960
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     protein_bind    1327..1360
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      complement(1369..1396)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(1412..1483)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    1531..1564
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(join(1581..1586,1587..1603,1604..1609))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1742..1759)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1876..2421)
                     /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(1956..1975)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2535..2629)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2660..3454)
                     /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(2771..2790)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3381..3400
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 cgaattcata tgactagtat cgttcgaaga tctaggtgcg cattcacagt tctccgcaag
       61 aattgattgg ctccaattct tggagtggtg aatccgttag cgaggtgccg ccggcttcca
      121 ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt
      181 atagggcggc gcctacaatc catgccaacc cgttccatgt gctcgccgag gcggcataaa
      241 tcgccgtgac gatcagcggt ccaatgatcg aagttaggct ggtaagagcc gcgagcgatc
      301 cttgaagctg tccctgatgg tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg
      361 gcatcccgat gccgccggaa gcgagaagaa tcataatggg gaaggccatc cagcctcgcg
      421 tcgcgaacgc cagcaagacg tagcccagcg cgtcggccgc catgccggcg ataatggcct
      481 gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca
      541 agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct
      601 cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga
      661 cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt
      721 tgaaggctct caagggcatc ggtcgacgct ctcccttatg cgactcctgc attaggaagc
      781 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
      841 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
      901 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
      961 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     1021 tccacaggac gggtgtggtc gccatgatcg cgtagtcgat agtggctcca agtagcgaag
     1081 cgagcaggac tgggcggcgg ccaaagcggt cggacagtgc tccgagaacg ggtgcgcata
     1141 gaaattgcat caacgcatat agcgctagca gcacgccata gtgactggcg atgctgtcgg
     1201 aatggacgat atcccgcaag aggcccggca gtaccggcat aaccaagcct atgcctacag
     1261 catccagggt gacggtgccg aggatgacga tgagcgcatt gttagatttc atacacgctt
     1321 cttaaaataa cttcgtataa tgtatgctat acgaagttat ggtataaacg cagaaaggcc
     1381 cacccgaagg tgagccagtg tgactctagt agagagcgtt caccgacaaa caacagataa
     1441 aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctg gagatcctta
     1501 ctcgagtttg gatccttatt tgatgcctgg ataacttcgt ataatgtatg ctatacgaag
     1561 ttatttcgcc gtgctaccgc attaaagctt atcgatgata agctgtcaaa catgagaatt
     1621 atcactagtg aatcatcgat ggtaccatat gttcgaagct tggcttgact ctcttccggg
     1681 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggagggata tattccgctt
     1741 cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg
     1801 aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa gtgagagggc
     1861 cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg aaatctgacg
     1921 ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1981 cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca ttccgctgtt
     2041 atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca gttcgctcca
     2101 agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta tccggtaact
     2161 atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca gccactggta
     2221 attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca
     2281 agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt ggtagctcag
     2341 agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca agagattacg
     2401 cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata tttctagatt
     2461 tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg
     2521 ttactagtgc ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac
     2581 aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc
     2641 tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct
     2701 gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa
     2761 gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca
     2821 gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc
     2881 aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg
     2941 cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa
     3001 gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg
     3061 ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt
     3121 tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca
     3181 gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg
     3241 tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca ccggacaggt
     3301 cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag
     3361 agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg
     3421 gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt
     3481 gatcagatca tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta
     3541 ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc gacgtcggtg
     3601 cctaatgagt gagctaactt acattaattg cgttgcgcat tcaccaccct gaa
//
