LOCUS       sequence-165717-        1039 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression of Cas9 and sgRNA scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1039)
  AUTHORS   Habashy et al
  TITLE     CRISPR/Cas9-based system for loss-of-function screening
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1039)
  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: 
            "Unpublished"
COMMENT     Sequence Label: lentiCRISPR v2-Blast
FEATURES             Location/Qualifiers
     source          1..1039
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(63..461)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(462..518)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(528..551)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(552..599)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 gtcaatcttt cacaaatttt gtaatccaga ggttgattac cgataagctt gatatcgaat
       61 tcttagccct cccacacata accagagggc agcaattcac gaatcccaac tgccgtcggc
      121 tgtccatcac tgtccttcac tatggctttg atcccaggat gcagatcgag aagcacctgt
      181 cggcaccgtc cgcaggggct caagatgccc ctgttctcat ttccgatcgc gacgatacaa
      241 gtcaggttgc cagctgccgc agcagcagca gtgcccagca ccacgagttc tgcacaaggt
      301 cccccagtaa aatgatatac attgacacca gtgaagatgc ggccgtcgct agagagagct
      361 gcgctggcga cgctgtagtc ttcagagatg gggatgctgt tgattgtagc cgttgctctt
      421 tcaatgaggg tggattcttc ttgagacaaa ggcttggcca tcggtccagg attctcttcg
      481 acatctccgg cttgtttcag cagagagaag tttgttgcgc cggatccctt atcgtcatcg
      541 tctttgtaat ctttcttctt cttagcctgt ccagccttct ttgtggcggc aggtcgcttg
      601 tcgcctccca gctgagacag gtcgatccgt gtctcgtaca ggccggtgat gctctggtgg
      661 atcagggtgg cgtccagcac ctctttggtg ctggtgtacc tcttccggtc gatggtggtg
      721 tcaaagtact tgaaggcggc aggggctccc agattggtca gggtaaacag gtggatgata
      781 ttctcggcct gctctctgat gggcttatcc cggtgcttgt tgtaggcgga cagcactttg
      841 tccagattag cgtcggccag gatcactctc ttggagaact cgctgatctg ctcgatgatc
      901 tcgtccaggt agtgcttgtg ctgttccaca aacagctgtt tctgctcatt atcctcgggg
      961 gagcccttca gcttctcata gtggctggcc aggtacagga agttcacata tttggagggc
     1021 agggccagtt cgtttccct
//
