LOCUS       sequence-112146-         893 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Lim Lab plasmid: AmpR + PLlacO-1::RBS(st7) gfp::Asp terminator + 
            ColE1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 893)
  AUTHORS   Sagawa S, Shin JE, Hussein R, Lim HN
  TITLE     Paradoxical suppression of small RNA activity at high Hfq 
            concentrations due to random-order binding.
  JOURNAL   Nucleic Acids Res. 2015 Aug 10. pii: gkv777.
  PUBMED    26261213
REFERENCE   2  (bases 1 to 893)
  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: "Nucleic 
            Acids Res. 2015 Aug 10. pii: gkv777."
COMMENT     Sequence Label: pHL1632
FEATURES             Location/Qualifiers
     source          1..893
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(22..735)
                     /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="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDW
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(62..82)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     677..705
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     protein_bind    763..779
                     /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 accctgtacg attaaagctt atttgtatag ttcatccatg ccatgtgtaa tcccagcagc
       61 tgttacaaac tcaagaagga ccatgtggtc tctcttttcg ttgggatctt tcgaaagggc
      121 agattgtgtg gacaggtaat ggttgtctgg taaaaggaca gggccatcgc caattggagt
      181 attttgttga taatggtctg ctagttgaac gcttccatct tcaatgttgt gtctaatttt
      241 gaagttaact ttgattccat tcttttgttt gtctgccatg atgtatacat tgtgtgagtt
      301 atagttgtat tccaatttgt gtccaagaat gtttccatct tctttaaaat caataccttt
      361 taactcgatt ctattaacaa gggtatcacc ttcaaacttg acttcagcac gtgtcttgta
      421 gttccagtca tctttgaaaa atatagttct ttcctgtaca taaccttcgg gcatggcact
      481 cttgaaaaag tcatgctgtt tcatatgatc tgggtatctc gcaaagcatt gaacaccata
      541 accgaaagta gtgacaagtg ttggccatgg aacaggtagt tttccagtag tgcaaataaa
      601 tttaagggta agttttccgt atgttgcatc accttcaccc tctccactga cagaaaattt
      661 gtgcccatta acatcaccat ctaattcaac aagaattggg acaactccag tgaaaagttc
      721 ttctccttta cgcatttttt ttcctcctta gtcgacagta tcttgttatc cgctcacaat
      781 gtcaatgtta tccgctcaca tttatgacgt caggtggcac ttttcgggga aatgtgcgcg
      841 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atg
//
