LOCUS       sequence-93085-e         893 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Lentiviral Vector for Sp sgRNA delivery without SpCas9, Puromycin 
            resistance, EFS Promoter driven.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 893)
  AUTHORS   Heckl D, Kowalczyk MS, Yudovich D, Belizaire R, Puram RV, McConkey 
            ME, Thielke A, Aster JC, Regev A, Ebert BL
  TITLE     Generation of mouse models of myeloid malignancy with combinatorial 
            genetic lesions using CRISPR-Cas9 genome editing.
  JOURNAL   Nat Biotechnol. 2014 Jun 22. doi: 10.1038/nbt.2951.
  PUBMED    24952903
REFERENCE   2  (bases 1 to 893)
  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: "Nat 
            Biotechnol. 2014 Jun 22. doi: 10.1038/nbt.2951."
COMMENT     Sequence Label: pLKO5.sgRNA.EFS.PAC
FEATURES             Location/Qualifiers
     source          1..893
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(41..640)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(124..144)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     621..640
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(666..877)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
ORIGIN
        1 aaattttgta atccagaggt tgattgtcga cttaacgcgt tcaggcaccg ggcttgcggg
       61 tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga
      121 gccgctcgta gaaggggagg ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg
      181 cgcgctcggc cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt
      241 ggtcgggcga gactccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg
      301 ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc
      361 gcgggccgat ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg
      421 ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga
      481 ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg acggtgtggc
      541 gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt
      601 cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat ggtggcggat cccgcgtcac
      661 gacacctgtg ttctggcggc aaacccgttg cgaaaaagaa cgttcacggc gactactgca
      721 cttatatacg gttctccccc accctcggga aaaaggcgga gccagtacac gacatcactt
      781 tcccagttta ccccgcgcca ccttctctag gcaccggttc aattgccgac ccctcccccc
      841 aacttctcgg ggactgtggg cgatgtgcgc tctgcccact gacgggcacc gga
//
