LOCUS       sequence-71644-e        1097 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 1097)
  TITLE     Series-9: N- or C-terminal Tag E.coli T7 Expression Vectors with 
            Biobrick Polycistronic Restriction Sites
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1097)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pET FLAG TEV cloning vector with BioBrick
            poly...
FEATURES             Location/Qualifiers
     source          1..1097
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     64..83
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        64..82
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             159..181
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             192..215
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             216..236
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(351..369)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      365..412
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(778..783,784..800,801..806))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(833..851)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        919..1023
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
       61 aattaatacg actcactata gggagaccac aacggtttcc ctctagtgcc ggctccggag
      121 agctctttaa tgcggccgct taattaacta gaaataattt tgtttaactt taagaaggag
      181 atatacatat ggactacaaa gacgatgacg acaaggaaaa cctgtacttc caatccaata
      241 ttggaagtgg ataacggatc gcgatcgccc tgcaggccgg ccggtaccac gcgtgcgcgc
      301 tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
      361 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
      421 aggaactata tccggatatc cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag
      481 tggctccaag tagcgaagcg agcaggactg ggcggcggcc aaagcggtcg gacagtgctc
      541 cgagaacggg tgcgcataga aattgcatca acgcatatag cgctagcagc acgccatagt
      601 gactggcgat gctgtcggaa tggacgatat cccgcaagag gcccggcagt accggcataa
      661 ccaagcctat gcctacagca tccagggtga cggtgccgag gatgacgatg agcgcattgt
      721 tagatttcat acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt
      781 aaagcttatc gatgataagc tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg
      841 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
      901 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
      961 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataacattg aaaaaggaag
     1021 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     1081 cctgtttttg ctcaccc
//
