LOCUS       sequence-194048-        1327 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Tetracycline inducible expression of GFPmut2 with constitutive TetR;
            moderate 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 1327)
  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 1327)
  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: pZH522
FEATURES             Location/Qualifiers
     source          1..1327
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     40..59
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      291..334
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(347..1060)
                     /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(387..407)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     1002..1030
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     protein_bind    1146..1164
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(1184..1203)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 annnnnnnnn annnnagnct gctctacacc tagcttctgg gcgagtttac gggttgttaa
       61 accttcgatt ccgacctcat taagcagctc taatgcgctg ttaatcactt tacttttatc
      121 taatctagac atggtgaaat cctcctaaag ttaaacaaaa ttatttgtag agggaaaccg
      181 ttgtggtctc cctgaatata tattacgagc cttatgcatg cccgtaaagt tatccagcaa
      241 ccactcatag acctagggca gcagataggg acgacgtggt gttagctgtg ataaaacgaa
      301 aggcccagtc tttcgactga gcctttcgtt ttatgctaga ttattatttg tatagttcat
      361 ccatgccatg tgtaatccca gcagctgtta caaactcaag aaggaccatg tggtctctct
      421 tttcgttggg atctttcgaa agggcagatt gtgtggacag gtaatggttg tctggtaaaa
      481 ggacagggcc atcgccaatt ggagtatttt gttgataatg gtctgctagt tgaacgcttc
      541 catcttcaat gttgtgtcta attttgaagt taactttgat tccattcttt tgtttgtctg
      601 ccatgatgta tacattgtgt gagttatagt tgtattccaa tttgtgtcca agaatgtttc
      661 catcttcttt aaaatcaata ccttttaact cgattctatt aacaagggta tcaccttcaa
      721 acttgacttc agcacgtgtc ttgtagttcc cgtcatcttt gaaaaatata gttctttcct
      781 gtacataacc ttcgggcatg gcactcttga aaaagtcatg ctgtttcata tgatctgggt
      841 atctcgcaaa gcattgaaga ccatacgcga aagtagtgac aagtgttggc catggaacag
      901 gtagttttcc agtagtgcaa ataaatttaa gggtaagttt tccgtatgtt gcatcacctt
      961 caccctctcc actgacagaa aatttgtgcc cattaacatc accatctaat tcaacaagaa
     1021 ttgggacaac tccagtgaaa agttcttctc ctttactcat gcggtacctt tctcggcttt
     1081 aatgaattcg gtcagtgcgt cctgctgaag tgctcagnat ccctatcact gnnaagggat
     1141 gtcaatctct atcactgata gggaactatg ggtgcactct cagtacaatc tgctctgatg
     1201 ccgcataatt aagccagccc cgaaacccgc caaaacccgg tgaacggccc tgaagggttn
     1261 nntcggctcc cggaatccgc ttaaaaaaca aannnnnnac cnnntcccgg gaatcnnnan
     1321 gggttca
//
