LOCUS       sequence-318615-        1325 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 1325)
  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 1325)
  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..1325
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(278..656)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     603..622
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(841..945)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1109..1130)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggngcctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
       61 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
      121 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
      181 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
      241 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
      301 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
      361 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
      421 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
      481 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
      541 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
      601 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
      661 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
      721 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
      781 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
      841 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
      901 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      961 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     1021 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     1081 tagaccgaaa atagggttga atgttgttcc agtttggaac aagagtccac tattaaagaa
     1141 cgtggactcc aacgtcaaag ggcaaaaaac cgtctatcag ggcgatggcc cactacgtga
     1201 accatccccc taatcaagtt ttttgggggg aagggggcca aaaaccacaa aacggaaccc
     1261 caaaggggnc ccccaaatta aaattggggg gaaacccnna aagggggaaa aaaaggaaaa
     1321 aacaa
//
