LOCUS       sequence-318618-        1212 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Modified lentiviral pKLV2-U6gRNA(BbsI)-PGKpuro2ABFP-W (#67974) 
            CRISPR gRNA expression vector which includes the ccdB gene flanked 
            by BbsI sites, to improve efficiency of oligonucleotide cloning..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1212)
  AUTHORS   Goncalves E, Thomas M, Behan FM, Picco G, Pacini C, Allen F, Vinceti
            A, Sharma M, Jackson DA, Price S, Beaver CM, Dovey O, Parry-Smith D,
            Iorio F, Parts L, Yusa K, Garnett MJ
  TITLE     Minimal genome-wide human CRISPR-Cas9 library.
  JOURNAL   Genome Biol. 2021 Jan 21;22(1):40. doi: 10.1186/s13059-021-02268-4.
  PUBMED    33478580
REFERENCE   2  (bases 1 to 1212)
  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: "Genome 
            Biol."; date: "2021-01-21"; volume: "22(1):40. doi"; pages: " 
            10.1186/s13059-021-02268-4"
COMMENT     Sequence Label: pKLV2-U6gRNA(BbsI)-ccdB-PGKpuro2ABFP-W
FEATURES             Location/Qualifiers
     source          1..1212
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(695..748)
                     /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"
     primer_bind     complement(847..867)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 catgctaagc cagtttgcta gggaggtcgc agtatctggc cactgccacc tcgtgctgct
       61 cgacgtaggt ctcgttgttg gcctccttga ttctttccag tctgtagtcc acatagtaga
      121 cgccaggcat cttgaggttc ttagcgggtt tcttggatct atatgtggtc ttgatgtttg
      181 cgatcagatg gctcccgccc acgagcttca gggccatgtc gtttctgcct tccaggccgc
      241 cgtcagcggg gtacagcgtc tcggtgaagg cctcccagcc gagtgttttc ttctgcatca
      301 cagggccgtt ggatgtgaag ttcacccctc tgatcttgac gttgtagatg aggcagccgt
      361 cctggaggct ggtgtcctgg gtagcggtca gcacgccccc gtcctcgtat gtggtgactc
      421 tctcccatgt gaagccctca gggaaggact gcttgaagaa gtcggggatg ccctgggtgt
      481 ggttgatgaa ggtcttgctg ccgtagagga agctagtagc caggatgtcg aaggcgaagg
      541 ggagagggcc gccctcgacc accttgattc tcatggtctg ggtgccctcg tagggcttgc
      601 cttcgccctc ggatgtgcac ttgaagtgat ggttgtccac ggtgccctcc atgtacagct
      661 tcatgtgcat gttctcctta atcagctcgc tcattgggcc aggattctcc tccacgtcac
      721 cgcatgttag gagacttcct ctgccctctc ctccggaccc gccgccggca ccgggcttgc
      781 gggtcatgca ccaggtgcgc ggtccttcgg gcacctcgac gtcggcggtg acggtgaagc
      841 cgagccgctc gtagaagggg aggttgcggg gcgcggaggt ctccaggaag gcgggcaccc
      901 cggcgcgctc ggccgcctcc actccgggga gcacgacggc gctgcccaga cccttgccct
      961 ggtggtcggg cgagacgccg acggtggcca ggaaccacgc gggctccttg ggccggtgcg
     1021 gcgccnggag gccttccatc tgttgctgcc cggccagccg ggaaccgctc aactcggcct
     1081 ggcgcgggcc gatctcggga aaaacggccc cggtttgaag ctttccgggg gggtccaaaa
     1141 cgccaccgcg gggccggcgt ccgggaacca aacctttggc aaagttcann ccnaaccccc
     1201 ttgaagaaaa at
//
