LOCUS       sequence-35255-e         872 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 872)
  TITLE     C-terminal fluorescent fusion vectors for E. coli expression
REFERENCE   2  (bases 1 to 872)
  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 Biotin His6 FlAsH LIC cloning vector (H6-F...
FEATURES             Location/Qualifiers
     source          1..872
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     52..71
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        52..70
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    71..95
                     /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             110..132
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             145..189
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             190..207
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             208..228
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             247..264
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     CDS             292..309
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(362..380)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      376..423
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      460..840
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(547..566)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     757..778
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa ttaatacgac
       61 tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact
      121 ttaagaagga gatataccat gggaggcctg aacgatattt ttgaagcgca gaaaattgaa
      181 tggcatgaac accatcacca tcaccatgaa aacctgtact tccaatccaa tattggtagt
      241 gggagctgtt gtcctggctg ttgcggtggc tagtaagcgg ccgcactcga gcaccaccac
      301 caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
      361 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
      421 ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat
      481 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
      541 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
      601 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
      661 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
      721 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
      781 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
      841 cctattggtt aaaaaatgag ctgatttaac aa
//
