LOCUS       sequence-35254-e         792 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 792)
  TITLE     C-terminal fluorescent fusion vectors for E. coli expression
REFERENCE   2  (bases 1 to 792)
  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"
COMMENT     Sequence Label: pET GFP domain 11 LIC cloning vector (GFP C-11)
FEATURES             Location/Qualifiers
     source          1..792
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     50..69
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        50..68
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    69..93
                     /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)."
     RBS             108..130
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             143..187
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             188..205
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             206..226
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             263..310
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             332..349
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(402..420)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      416..463
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(587..606)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc
       61 actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt
      121 aagaaggaga tataccatgg gaggcctgaa cgatattttt gaagcgcaga aaattgaatg
      181 gcatgaacac catcaccatc accatgaaaa cctgtacttc caatccaata ttggtagtgg
      241 gagcaacggc agcagcggat cccgcgatca catggtcctg cacgagtacg tgaacgccgc
      301 cgggatcact tagtaagcgg ccgcactcga gcaccaccac caccaccact gagatccggc
      361 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
      421 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
      481 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
      541 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
      601 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
      661 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
      721 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
      781 tccacgttct tt
//
