LOCUS       sequence-83568-e         891 bp    DNA     linear   UNA 15-APR-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 891)
  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 891)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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..891
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             7..29
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             48..65
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             75..92
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(144..470)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRFKQSTLDLIKDPARPRVPSSNSLAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGVSSGN
                     LPQL"
     primer_bind     338..360
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     352..369
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     353..369
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(466..482)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(466..482)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(479..501)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    490..506
                     /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)."
     promoter        complement(join(514..520,521..538,539..544))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    559..580
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             739..756
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(809..827)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      823..870
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 aataattttg tttaacttta agaaggagat ataccatggg cagcagccat catcatcatc
       61 atcacagcag cggcctggtg ccgcgcggca gccatggccg cagccgcctt ttggaagatt
      121 ttcgaaacaa ccggtacccc aatttacaac tgcgggagat tgccggaaga gaccccgcac
      181 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
      241 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      301 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      361 acggccagtg aattcgagct cggtacccgg gggcgcgccg gatccttaat taagtctaga
      421 gtcgactgtt taaacctgca ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc
      481 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
      541 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
      601 cgctttccag tcggtctcaa ccccacatcg caactcttcg taagtacacc tatggcaagc
      661 acattctggc caagctggag aagtactaca tgaagaacgg tgttgactta gggtaacaag
      721 cttgcggccg cactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc
      781 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
      841 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc c
//
