LOCUS       sequence-263853-        2830 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses the full-length Beclin-1 protein (a.a. 1-450) with a 
            N-terminal hexa-Histidine and solubility tag (i.e., Trigger Factor).
            Three mutations are added to the CC and ECD/BARA interface..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2830)
  AUTHORS   Ranaghan MJ, Durney MA, Mesleh MF, McCarren PR, Garvie CW, Daniels 
            DS, Carey KL, Skepner AP, Levine B, Perez JR
  TITLE     The Autophagy-Related Beclin-1 Protein Requires the Coiled-Coil and 
            BARA Domains To Form a Homodimer with Submicromolar Affinity.
  JOURNAL   Biochemistry. 2017 Dec 26;56(51):6639-6651. doi: 
            10.1021/acs.biochem.7b00936. Epub 2017 Dec 14.
  PUBMED    29185708
REFERENCE   2  (bases 1 to 2830)
  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"; doi: 
            "10.1021/acs.biochem.7b00936"; journalName: "Biochemistry"; date: 
            "2017-12-26- 26"; volume: "56"; issue: "51"; pages: "6639-6651"
COMMENT     Sequence Label: BPV00914 pET21GG2 H6-TF-BECN (1-450) [VA-MA-LA]
FEATURES             Location/Qualifiers
     source          1..2830
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             95..112
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1433..1450
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1457..1477
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     regulatory      2580..2589
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gggtcatcat catcaccatc atggtagcat
      121 gcaggttagc gttgaaacca cccagggtct gggtcgtcgt gttaccatta ccattgcagc
      181 agatagcatt gaaaccgcag ttaaaagcga actggttaac gttgcaaaaa aagtgcgcat
      241 tgatggtttt cgcaaaggta aagttccgat gaatattgtt gcacagcgtt atggtgcaag
      301 cgttcgtcag gatgttctgg gtgatctgat gagccgtaac tttattgatg ccatcatcaa
      361 agaaaaaatc aatccggcag gcgcaccgac ctatgttccg ggtgaatata aactgggtga
      421 agattttacc tacagcgtgg aatttgaagt ttatccggaa gttgaactgc agggcctgga
      481 agcaattgaa gttgaaaaac cgattgttga agttaccgat gcagatgttg atggtatgct
      541 ggataccctg cgtaaacagc aggcaacctg gaaagaaaaa gatggtgcag ttgaagccga
      601 agatcgtgtg accattgatt tcaccggtag cgttgatggt gaagaatttg agggtggtaa
      661 agcaagcgat tttgttctgg caatgggtca gggtcgtatg attccgggtt ttgaagatgg
      721 tattaaaggt cataaagccg gtgaggaatt taccattgat gttacctttc cggaagaata
      781 tcatgccgaa aatctgaaag gcaaagcagc caaatttgcc atcaatctga aaaaagtgga
      841 agaacgtgaa ctgccggaac tgaccgcaga atttatcaaa cgttttggtg tggaagatgg
      901 cagcgttgaa ggtctgcgtg ccgaagttcg caaaaatatg gaacgcgaac tgaaaagcgc
      961 aattcgtaat cgtgttaaaa gccaggccat tgaaggcctg gttaaagcaa atgatattga
     1021 tgttcctgca gccctgattg atagcgaaat tgatgtgctg cgtcgtcagg cagcccagcg
     1081 ctttggtggt aatgaaaaac aggcactgga actgcctcgt gaactgtttg aagaacaggc
     1141 aaaacgtcgt gttgttgttg gtctgctgct gggcgaagtt attcgtacca atgaactgaa
     1201 agcagatgaa gaacgcgtga aaggtctgat tgaagaaatg gcaagcgcct atgaagatcc
     1261 gaaagaagtg attgagttct atagcaaaaa caaagaactg atggacaaca tgcgtaatgt
     1321 tgcactggaa gaacaagccg ttgaagcagt tctggccaaa gcaaaagtga ccgaaaaaga
     1381 aaccaccttt aacgaactga tgaatcagca agccggtgca agcaccggat ccctggttcc
     1441 gcgtggtagc gctagcgaaa atctgtattt tcagggaatg gaaggtagca aaaccagcaa
     1501 taatagcacc atgcaggtta gctttgtttg tcagcgttgt agccagccgc tgaaactgga
     1561 taccagcttt aaaatcctgg atcgtgttac cattcaagaa ctgaccgcac cgctgctgac
     1621 caccgcacag gcaaaaccgg gtgaaaccca agaagaagaa accaattcag gcgaagaacc
     1681 gtttattgaa acaccgcgtc aggatggtgt tagccgtcgt tttattccgc ctgcacgtat
     1741 gatgagcacc gaaagcgcaa atagctttac cctgattggt gaagcaagtg atggtggcac
     1801 catggaaaat ctgagccgtc gtctgaaagt taccggtgac ctgtttgata tcatgagcgg
     1861 tcagaccgat gttgatcatc cgctgtgtga agaatgtacc gataccctgc tggatcagct
     1921 ggatacccaa ctgaatgtta ccgaaaatga atgccagaac tataaacgct gtctggaaat
     1981 tctggaacaa atgaacgaag atgatagcga acagctgcag atggaactga aagaactggc
     2041 actggaagaa gaacgtctga tccaagaact ggaagatgtg gaaaaaaacc gtaaaattgt
     2101 ggccgagaat ctggaaaaag ttcaggcaga agccgaacgc ctggatcaag aagaggcaca
     2161 gtatcagcgt gaatatagcg aatttaaacg tcagcagctg gaactggacg atgaactgaa
     2221 aagcgctgaa aatcaggcgc gttatgcaca gacccaggcg gacaaactga aaaaaacgaa
     2281 tgtttttaac gccacctttc acatttggca tagtggtcag tttggcacca ttaataactt
     2341 tcgtctgggt cgtctgccga gcgttccggt tgaatggaat gaaattaatg cagcatgggg
     2401 tcagacagtt ctgctgctgc atgcactggc caataaaatg ggtctgaaat tccagcgtta
     2461 tcgtctggtt ccgtatggta atcatagcta tctggaaagc ctgaccgata aaagcaaaga
     2521 actgccgctg tattgtagcg gtggtctgcg ttttttttgg gataacaaat ttgatcatgc
     2581 catggtggca tttctggatt gtgttcagca gtttaaagaa gaggttgaaa aaggcgaaac
     2641 ccgtttttgt ctgccgtatc gtatggatgt agaaaaaggc aaaattgaag ataccggtgg
     2701 tagtggtggt agctatagca ttaaaaccca gtttaatagc gaagaacagt ggaccaaagc
     2761 cctgaaattt atgctgacca atctgaaatg gggtctggca tgggttagca gccagtttta
     2821 taacaaataa
//
