LOCUS       sequence-318616-        1418 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 1418)
  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 1418)
  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..1418
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             40..828
                     /codon_start=1
                     /gene="bla(M)"
                     /product="beta-lactamase lacking the signal sequence"
                     /label=bla(M)
                     /note="allows cytosolic expression of beta-lactamase"
                     /translation="PETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMST
                     FKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMS
                     DNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATT
                     LRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAA
                     LGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     primer_bind     complement(186..205)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
       61 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
      121 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
      181 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
      241 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
      301 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
      361 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
      421 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
      481 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
      541 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
      601 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
      661 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
      721 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
      781 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
      841 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
      901 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
      961 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1021 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1081 caagagctac caactctttt tccgaaggta actggcttca gcagaacgca gaaaccaaat
     1141 actgttcttc tagggaagcc gtagttaggc caccacttca agaactctgt aacaccgcca
     1201 aatacctcgc ttggcaatcc cggttcccgg ggnngcggcc cgggggaaaa aatcgggnnn
     1261 ttacccgggt tggaccaaaa aaaaaatann naaaaaggcn cccgnnnggg nnnaaaaggg
     1321 gggtttgggc acccccnnnn tttgaaaaaa aaacacaacc caaannnaaa aataaanngg
     1381 ggnntttaaa aannnccccc ctcccccaag gaaaagag
//
