LOCUS       sequence-194042-        1518 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Constitutive expression of GFPmut2; 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 1518)
  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 1518)
  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: pZH516
FEATURES             Location/Qualifiers
     source          1..1518
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    35..53
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             113..826
                     /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(143..171)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     766..786
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      862..908
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(927..943)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(927..943)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(940..962)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    951..967
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(975..981,982..999,1000..1005))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1020..1041
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 annnncnnat gtcnatcagt gatagagatt gacatcccta tcagtgatag agatactgag
       61 cacatcagca ggacgcactg accgaattca ttaaagccga gaaaggtacc gcatgagtaa
      121 aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa
      181 tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac
      241 ccttaaattt atttgcacta ctggaaaact acctgttcca tggccaacac ttgtcactac
      301 tttcgcgtat ggtcttcaat gctttgcgag atacccagat catatgaaac agcatgactt
      361 tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga
      421 cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat
      481 cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttggacaca aattggaata
      541 caactataac tcacacaatg tatacatcat ggcagacaaa caaaagaatg gaatcaaagt
      601 taacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag accattatca
      661 acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac
      721 acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt
      781 tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaataat aatctagcca
      841 tggtacgcgt gctagaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
      901 cgttttataa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
      961 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1021 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1081 ctgtcgtgcc agctgcatta atgaatcggc caacgccccg ggaaaaggcg gtttgcgtat
     1141 tggggcgctc ttccgcttcc tcgttacatg aatncntggc cnngnttcgt tcggctgcgg
     1201 ggaaacggta tcagctcact caaaggccgg taataccggt tatcccanga aatccggggg
     1261 ataaccccag gaaaaaacat tgnnnaccaa aaggccccnn aaaagggccc gaaaacnnna
     1321 aaaaggcgcc cctacgcgaa aggnaaantg ggcttannnn nntnggccac ntagagaggg
     1381 tgtcccaaaa aaagncttnn ttngtnnnnn nngngaagaa ggcgccctcn gggggccnnc
     1441 nanaaaannn nnnntaannn nnngaatatt nccccnttcn nnnnccnaga tacanntcna
     1501 acaaatnccn tnnnncaa
//
