LOCUS       sequence-80790-e         793 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Receiving vector for PUM-HD assembly and expression in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 793)
  AUTHORS   Abil Z, Denard CA, Zhao H
  TITLE     Modular assembly of designer PUF proteins for specific 
            post-transcriptional regulation of endogenous RNA.
  JOURNAL   J Biol Eng. 2014 Mar 1;8(1):7.
  PUBMED    24581042
REFERENCE   2  (bases 1 to 793)
  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: "J Biol 
            Eng."; date: "2014-03-1"; volume: "8(1)"; pages: "7"
COMMENT     Sequence Label: pET28-GG-PUF
FEATURES             Location/Qualifiers
     source          1..793
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(47..64)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(47..63)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(56..78)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(324..341)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(351..368)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             387..409
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    424..448
                     /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)."
     primer_bind     complement(448..467)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(449..467)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(551..570)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 acttaattaa ggatccggcg cgcccccggg taccgagctc gaattcactg gccgtcgttt
       61 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
      121 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
      181 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
      241 gggtctcttc cggcaatctc ccgcagttgt aaattggggt accggttgtt tcgaaaatct
      301 tccaaaaggc ggctgcggcc atggctgccg cgcggcacca ggccgctgct gtgatgatga
      361 tgatgatggc tgctgcccat ggtatatctc cttcttaaag ttaaacaaaa ttatttctag
      421 aggggaattg ttatccgctc acaattcccc tatagtgagt cgtattaatt tcgcgggatc
      481 gagatctcga tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg
      541 gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg
      601 ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg
      661 ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta
      721 ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgaga tcccggacac
      781 catcgaatgg cgc
//
