LOCUS       sequence-90412-e         827 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 827)
  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 827)
  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..827
                     /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             complement(140..466)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRFKQSTLDLIKDPARPRVPSSNSLAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGVSSGN
                     LPQL"
     primer_bind     334..356
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     348..365
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     349..365
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(462..478)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(462..478)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(475..497)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    486..502
                     /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(510..516,517..534,535..540))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    555..576
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             735..752
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(805..823)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 attttgttta actttaagaa ggagatatac catgggcagc agccatcatc atcatcatca
       61 cagcagcggc ctggtgccgc gcggcagcca tggccgcagc cgccttttgg aagattttcg
      121 aaacaaccgg taccccaatt tacaactgcg ggagattgcc ggaagagacc ccgcacagat
      181 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
      241 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
      301 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
      361 ccagtgaatt cgagctcggt acccgggggc gcgccggatc cttaattaag tctagagtcg
      421 actgtttaaa cctgcaggca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg
      481 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
      541 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
      601 ttccagtcgg tctcaacccc acatcgcaac tcttcgtaag tacacctatg gcaagcacat
      661 tctggccaag ctggagaagt actacatgaa gaacggtgtt gacttagggt aacaagcttg
      721 cggccgcact cgagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa
      781 aggaagctga gttggctgct gccaccgctg agcaataact agcataa
//
