LOCUS       sequence-194046-        1396 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Tetracycline inducible expression of GFPmut2 with constitutive TetR;
            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 1396)
  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 1396)
  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: pZH520
FEATURES             Location/Qualifiers
     source          1..1396
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     25..44
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      276..319
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(332..1045)
                     /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(372..392)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     987..1015
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     RBS             1054..1065
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 angctgctct acacctagct tctgggcgag tttacgggtt gttaaacctt cgattccgac
       61 ctcattaagc agctctaatg cgctgttaat cactttactt ttatctaatc tagacatggt
      121 gaaatcctcc taaagttaaa caaaattatt tgtagaggga aaccgttgtg gtctccctga
      181 atatatatta cgagccttat gcatgcccgt aaagttatcc agcaaccact catagaccta
      241 gggcagcaga tagggacgac gtggtgttag ctgtgataaa acgaaaggcc cagtctttcg
      301 actgagcctt tcgttttatg ctagattatt atttgtatag ttcatccatg ccatgtgtaa
      361 tcccagcagc tgttacaaac tcaagaagga ccatgtggtc tctcttttcg ttgggatctt
      421 tcgaaagggc agattgtgtg gacaggtaat ggttgtctgg taaaaggaca gggccatcgc
      481 caattggagt attttgttga taatggtctg ctagttgaac gcttccatct tcaatgttgt
      541 gtctaatttt gaagttaact ttgattccat tcttttgttt gtctgccatg atgtatacat
      601 tgtgtgagtt atagttgtat tccaatttgt gtccaagaat gtttccatct tctttaaaat
      661 caataccttt taactcgatt ctattaacaa gggtatcacc ttcaaacttg acttcagcac
      721 gtgtcttgta gttcccgtca tctttgaaaa atatagttct ttcctgtaca taaccttcgg
      781 gcatggcact cttgaaaaag tcatgctgtt tcatatgatc tgggtatctc gcaaagcatt
      841 gaagaccata cgcgaaagta gtgacaagtg ttggccatgg aacaggtagt tttccagtag
      901 tgcaaataaa tttaagggta agttttccgt atgttgcatc accttcaccc tctccactga
      961 cagaaaattt gtgcccatta acatcaccat ctaattcaac aagaattggg acaactccag
     1021 tgaaaagttc ttctccttta ctcatgcggt acctttctcc tctttaatga attcggtcag
     1081 tgcgtcctgc tgatgtgctc agtatctcta tcactgaaan gggatgtcaa tctctatcac
     1141 tgaaagggac atatgggtgc annnccagta caatctgctc tgaaggcgna aaattnaacc
     1201 cagccccnaa cacccgccaa aaaccggctg aacgggccct gaagggtttn gtcggctccc
     1261 ggtatccgct taaaaaaaaa acttgggacc gttccccggg ntctngnntg gggtcaaaag
     1321 gttttccccc gccccccccc aaaacnnnnn gaaaaaaaaa gggcctnnng atnnccccnt
     1381 aattttnttg aggtnt
//
