LOCUS       Exported                 854 bp ds-DNA     linear   SYN 17-MAY-2021
DEFINITION  Express sgRNA and dCas9-KRAB from lentiviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    pLV hU6-sgRNA hUbC-dCas9-KRAB-T2a-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 854)
  AUTHORS   Thakore PI, D'Ippolito AM, Song L, Safi A, Shivakumar NK, Kabadi AM,
            Reddy TE, Crawford GE, Gersbach CA
  TITLE     Highly specific epigenome editing by CRISPR-Cas9 repressors for 
            silencing of distal regulatory elements.
  JOURNAL   Nat Methods. 2015 Dec;12(12):1143-9. doi: 10.1038/nmeth.3630. Epub 
            2015 Oct 26.
  PUBMED    26501517
REFERENCE   2  (bases 1 to 854)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 17, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..854
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     441..460
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(461..514)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(521..541)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(608..802)
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQILYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
ORIGIN
        1 cgcgctcggc cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt
       61 ggtcgggcga cacgccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg
      121 ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc
      181 gcgggccgat ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg
      241 ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga
      301 ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg acggtgtggc
      361 gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt
      421 cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat agggccggga ttctcctcca
      481 cgtcaccgca tgttagaaga cttcctctgc cctcgctagc aactttgcgt ttctttttcg
      541 gaactgatga tttgatttca aatgcagtct ctgaatcagg atgggtctct tggtgaattt
      601 ctctctccac cagccagggc tcttctccct tctccaaccg gaggatcaca tctggcttag
      661 taagctgata acccaaggaa accaggttct tatagttctc cagcatcaca tttctgtaca
      721 ggatctgctg agcagtgtcc agcagcttcc actcctccct ggtgaagtcc acaaacacat
      781 ccttgaaggt caccagtgtc cgggaccagg cagtcagtga cttagcatcg ctagccacct
      841 tcctcttctt cttg
//
