LOCUS       sequence-22954-e         939 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 939)
  TITLE     Other Plasmids
REFERENCE   2  (bases 1 to 939)
  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: M62 pPET28a-TEV-human beta cat domain N(1-137)
FEATURES             Location/Qualifiers
     source          1..939
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             3..25
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             44..61
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             71..88
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             92..124
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             134..154
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             200..235
                     /codon_start=1
                     /product="nanobody epitope tag corresponding to amino acids
                     16-27 of human beta-catenin (Traenkle et al., 2015)"
                     /label=BC2 tag
                     /translation="PDRKAAVSHWQQ"
     CDS             596..613
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(666..684)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      680..727
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(851..870)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 attttgttta actttaagaa ggagatatac catgggcagc agccatcatc atcatcatca
       61 cagcagcggc ctggtgccgc gcggcagcca tatggctagc atgactggtg gacagcaaat
      121 gggtcgcgga tccgaaaacc tgtattttca gggcatggct actcaagctg atttgatgga
      181 gttggacatg gccatggaac cagacagaaa agcggctgtt agtcactggc agcaacagtc
      241 ttacctggac tctggaatcc attctggtgc cactaccaca gctccttctc tgagtggtaa
      301 aggcaatcct gaggaagagg atgtggatac ctcccaagtc ctgtatgagt gggaacaggg
      361 attttctcag tccttcactc aagaacaagt agctgatatt gatggacagt atgcaatgac
      421 tcgagctcag agggtacgag ctgctatgtt ccctgagaca ttagatgagg gcatgcagat
      481 cccatctaca cagtttgatg ctgctcatcc cactaatgtc cagcgtttgg ctgaaccatc
      541 acagatgctg aaacatgcag ttgtataagt cgacaagctt gcggccgcac tcgagcacca
      601 ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
      661 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
      721 ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc
      781 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
      841 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg cncgtcgcnn ttccnnagnn
      901 natnnagctc naaatcgggg gctcccttta gggttccga
//
