LOCUS       sequence-194040-        1375 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Constitutive expression of GFPmut2; 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 1375)
  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 1375)
  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: pZH514
FEATURES             Location/Qualifiers
     source          1..1375
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    36..54
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             94..105
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             114..827
                     /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(144..172)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     767..787
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      863..909
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(928..944)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(928..944)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(941..963)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    952..968
                     /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)."
ORIGIN
        1 angnnnntnn gnccnatcag tgatagagat tgacatccct atcagtgata gagatactga
       61 gcacatcagc aggacgcact gaccgaattc attaaagagg agaaaggtac cgcatgagta
      121 aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta
      181 atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac ggaaaactta
      241 cccttaaatt tatttgcact actggaaaac tacctgttcc atggccaaca cttgtcacta
      301 ctttcgcgta tggtcttcaa tgctttgcga gatacccaga tcatatgaaa cagcatgact
      361 ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg
      421 acgggaacta caagacacgt gctgaagtca agtttgaagg tgataccctt gttaatagaa
      481 tcgagttaaa aggtattgat tttaaagaag atggaaacat tcttggacac aaattggaat
      541 acaactataa ctcacacaat gtatacatca tggcagacaa acaaaagaat ggaatcaaag
      601 ttaacttcaa aattagacac aacattgaag atggaagcgt tcaactagca gaccattatc
      661 aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat tacctgtcca
      721 cacaatctgc cctttcgaaa gatcccaacg aaaagagaga ccacatggtc cttcttgagt
      781 ttgtaacagc tgctgggatt acacatggca tggatgaact atacaaataa taatctagcc
      841 atggtacgcg tgctagaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
      901 tcgttttata agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
      961 gctcacaatt ccacacaaca tacgaacccg gaagcataaa gtgtaaagcc tgggggtgcc
     1021 taanggagtg gagctaaact cacattaaat tgccgtggcg ctcactggcc cgcctttcca
     1081 gtccggaaaa cctggtcgtg gccagctgcc attaatgaan cnggccaacg ccccggggaa
     1141 angncggttt tgcnttattg gacngctctt tnngattctc tcgctttaac ggaatccgtt
     1201 gggtacccgt ccntttcggt ctgcngnnna atcggatttc atcntccctc caaaaggcgg
     1261 gtaaaaacgg gttanacnnn taaaaatcca ggtgtaaaan acccaggaaa aagacctgtg
     1321 gaacaactaa atgncccctc aaaaaagtcc aggataactg tataaannan cttgg
//
