LOCUS       sequence-90372-e        2976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of APX W41F under control of TetR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2976)
  AUTHORS   Dwyer DJ, Belenky PA, Yang JH, MacDonald IC, Martell JD, Takahashi 
            N, Chan CT, Lobritz MA, Braff D, Schwarz EG, Ye JD, Pati M, 
            Vercruysse M, Ralifo PS, Allison KR, Khalil AS, Ting AY, Walker GC, 
            Collins JJ
  TITLE     Antibiotics induce redox-related physiological alterations as part 
            of their lethality.
  JOURNAL   Proc Natl Acad Sci U S A. 2014 May 20;111(20):E2100-9. doi: 
            10.1073/pnas.1401876111. Epub 2014 May 6.
  PUBMED    24803433
REFERENCE   2  (bases 1 to 2976)
  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"; doi: 
            "10.1073/pnas.1401876111"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2014-05-20- 20"; volume: "111"; issue: "20"; pages: "E2100-9"
COMMENT     Sequence Label: pZE21-APX(W41F) dimeric
FEATURES             Location/Qualifiers
     source          1..2976
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(7..25,26..31,32..48,49..54,55..80)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    7..25
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    32..50
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             90..101
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      920..1006
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1170..1758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1250..1269)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1846..1940)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1971..2765)
                     /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(2082..2101)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2692..2711
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2944..2962
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagtccc tatcagtgat agagattgac atccctatca gtgatagaga tactgagcac
       61 atcagcagga cgcactgacc gaattcatta aagaggagaa aggtaccatg ggaaagtcat
      121 acccaaccgt gagcccagat taccagaagg ctattgagaa ggcaaaaagg aaactgaggg
      181 gattcatcgc agaaaagaaa tgcgccccac tgatcctgcg actggctttc cacagcgcag
      241 gaacatttga ctccaagacc aaaacaggcg gaccctttgg gactatcaag caccaggctg
      301 aactggcaca tggggccaac aatggcctgg acattgctgt gagactgctg gagccaatca
      361 aagaacagtt ccccattgtc tcctacgcag acttctacca gctggccggc gtggtcgctg
      421 tggagattac cggcggcccc gaggtccctt tccacccagg aagggaggac aaacctgaac
      481 cccctccaga ggggagactg ccagatgcta ccaagggcag cgaccatctg cgggacgtgt
      541 tcggaaaagc aatggggctg tctgaccagg atatcgtggc cctgagtgga gggcacacca
      601 ttggcgccgc tcataaggaa cgctcaggct tcgagggacc ctggacaagc aaccctctga
      661 tcttcgacaa ttcctacttt actgaactgc tgaccggcga gaaggatgga ctgctgcagc
      721 tgccctctga caaagccctg ctgacagata gcgtgttcag gcctctggtc gaaaagtatg
      781 cagccgacga ggatgtgttc ttcgccgact atgctgaggc acacctgaaa ctgtctgaac
      841 tgggattcgc cgaggcatag aagcttgata tcgaattcct gcagcccggg ggatcccatg
      901 gtacgcgtgc tagaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
      961 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgccctagac
     1021 ctaggcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     1081 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1141 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1201 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1261 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1321 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     1381 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1441 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1501 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1561 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     1621 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     1681 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     1741 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     1801 aacgaaaact cacgttaagg gattttggtc atgactagtg cttggattct caccaataaa
     1861 aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac
     1921 tggatctatc aacaggagtc caagcgagct ctcgaacccc agagtcccgc tcagaagaac
     1981 tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc
     2041 acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac
     2101 gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag
     2161 cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc
     2221 tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga
     2281 tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc
     2341 tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc
     2401 cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg
     2461 agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg
     2521 tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg
     2581 tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc
     2641 tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca
     2701 tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca
     2761 atcatgcgaa acgatcctca tcctgtctct tgatcagatc ttgatcccct gcgccatcag
     2821 atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag
     2881 ggcgccccag ctggcaattc cgacgtctaa gaaaccatta ttatcatgac attaacctat
     2941 aaaaataggc gtatcacgag gccctttcgt cttcac
//
