LOCUS       sequence-194047-        1345 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Tetracycline inducible expression of GFPmut2 with constitutive TetR;
            weak 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 1345)
  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 1345)
  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: pZH521
FEATURES             Location/Qualifiers
     source          1..1345
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     23..42
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      274..317
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(330..1043)
                     /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(370..390)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     985..1013
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     promoter        complement(join(1073..1098,1099..1104,1105..1121,1122..1127,
                     1128..1146))
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1103..1121
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1128..1146
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(1165..1184)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gctgctctac acctagcttc tgggcgagtt tacgggttgt taaaccttcg attccgacct
       61 cattaagcag ctctaatgcg ctgttaatca ctttactttt atctaatcta gacatggtga
      121 aatcctccta aagttaaaca aaattatttg tagagggaaa ccgttgtggt ctccctgaat
      181 atatattacg agccttatgc atgcccgtaa agttatccag caaccactca tagacctagg
      241 gcagcagata gggacgacgt ggtgttagct gtgataaaac gaaaggccca gtctttcgac
      301 tgagcctttc gttttatgct agattattat ttgtatagtt catccatgcc atgtgtaatc
      361 ccagcagctg ttacaaactc aagaaggacc atgtggtctc tcttttcgtt gggatctttc
      421 gaaagggcag attgtgtgga caggtaatgg ttgtctggta aaaggacagg gccatcgcca
      481 attggagtat tttgttgata atggtctgct agttgaacgc ttccatcttc aatgttgtgt
      541 ctaattttga agttaacttt gattccattc ttttgtttgt ctgccatgat gtatacattg
      601 tgtgagttat agttgtattc caatttgtgt ccaagaatgt ttccatcttc tttaaaatca
      661 atacctttta actcgattct attaacaagg gtatcacctt caaacttgac ttcagcacgt
      721 gtcttgtagt tcccgtcatc tttgaaaaat atagttcttt cctgtacata accttcgggc
      781 atggcactct tgaaaaagtc atgctgtttc atatgatctg ggtatctcgc aaagcattga
      841 agaccatacg cgaaagtagt gacaagtgtt ggccatggaa caggtagttt tccagtagtg
      901 caaataaatt taagggtaag ttttccgtat gttgcatcac cttcaccctc tccactgaca
      961 gaaaatttgt gcccattaac atcaccatct aattcaacaa gaattgggac aactccagtg
     1021 aaaagttctt ctcctttact catgcggtac ctttctcgaa tttaatgaat tcggtcagtg
     1081 cgtcctgctg atgtgctcag tatctctatc actgataggg atgtcaatct ctatcactga
     1141 tagggacata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     1201 ccgacacccg ccaaaacccg gtgaacggcc ctgaagggtt tgtcggctcc ggcatccgct
     1261 taannaaaaa cttggaccgt ctccgggaac ttcatggtgc aagaggtttc cccnncatca
     1321 ccaaaacccc cnnaacaaaa ggtca
//
