LOCUS       sequence-194043-        1339 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Tetracycline inducible expression of GFPmut2 using autoregulation; 
            strong ribosome binding site; GFPmut2 ORF can be replaced with gene 
            of interest by isothermal assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1339)
  AUTHORS   Hensel Z
  TITLE     A plasmid-based Escherichia coli gene expression system with 
            cell-to-cell variation below the extrinsic noise limit.
  JOURNAL   PLoS One. 2017 Oct 30;12(10):e0187259. doi: 
            10.1371/journal.pone.0187259. eCollection 2017.
  PUBMED    29084263
REFERENCE   2  (bases 1 to 1339)
  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: "PLoS One.";
            date: "2017-10-30"; volume: "12(10):e0187259. doi"; pages: " 
            10.1371/journal.pone.0187259. eCollection 2017"
COMMENT     Sequence Label: pZH517
FEATURES             Location/Qualifiers
     source          1..1339
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     gap             2..12
                     /estimated_length=11
     primer_bind     35..54
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        complement(join(142..167,168..173,174..190,191..196,
                     197..215))
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    172..190
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    197..215
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      216..259
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(272..985)
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(312..332)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     927..955
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     RBS             994..1005
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(join(1015..1040,1041..1046,1047..1063,1064..1069,
                     1070..1088))
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1045..1063
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1070..1088
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(1107..1126)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 annnnnnnnn nngctgctct acacctagct tctgggcgag tttacgggtt gttaaacctt
       61 cgattccgac ctcattaagc agctctaatg cgctgttaat cactttactt ttatctaatc
      121 tagacatggt gaaatcctcc tggtcagtgc gtcctgctga tgtgctcagt atctctatca
      181 ctgataggga tgtcaatctc tatcactgat agggaataaa acgaaaggcc cagtctttcg
      241 actgagcctt tcgttttatg ctagattatt atttgtatag ttcatccatg ccatgtgtaa
      301 tcccagcagc tgttacaaac tcaagaagga ccatgtggtc tctcttttcg ttgggatctt
      361 tcgaaagggc agattgtgtg gacaggtaat ggttgtctgg taaaaggaca gggccatcgc
      421 caattggagt attttgttga taatggtctg ctagttgaac gcttccatct tcaatgttgt
      481 gtctaatttt gaagttaact ttgattccat tcttttgttt gtctgccatg atgtatacat
      541 tgtgtgagtt atagttgtat tccaatttgt gtccaagaat gtttccatct tctttaaaat
      601 caataccttt taactcgatt ctattaacaa gggtatcacc ttcaaacttg acttcagcac
      661 gtgtcttgta gttcccgtca tctttgaaaa atatagttct ttcctgtaca taaccttcgg
      721 gcatggcact cttgaaaaag tcatgctgtt tcatatgatc tgggtatctc gcaaagcatt
      781 gaagaccata cgcgaaagta gtgacaagtg ttggccatgg aacaggtagt tttccagtag
      841 tgcaaataaa tttaagggta agttttccgt atgttgcatc accttcaccc tctccactga
      901 cagaaaattt gtgcccatta acatcaccat ctaattcaac aagaattggg acaactccag
      961 tgaaaagttc ttctccttta ctcatgcggt acctttctcc tctttaatga attcggtcag
     1021 tgcgtcctgc tgatgtgctc agtatctcta tcactgatag ggatgtcaat ctctatcact
     1081 gatagggaca tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     1141 ccccgacacc cgccaaaacc cgctgaagcg ccctgaaggg cttgtctgct cccggcatcc
     1201 gcttacagac aagcttggac cgtctccggg aactgcatgg gtcaaaagtt ttcaccgtca
     1261 tcaccgaaac gccnnngacg aaanggcctn gggaaacccc nttttnnnna aggtgaagnn
     1321 ngnnaaaaaa aagggtttc
//
