LOCUS       sequence-146452-         940 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  lenti sgRNA cloning backbone with MS2 loops at tetraloop and 
            stemloop 2, dCas9-VP64, and blast resistance marker. Contains BsmBI 
            sites for insertion of spacer sequences..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 940)
  AUTHORS   Joung J, Konermann S, Gootenberg JS, Abudayyeh OO, Platt RJ, Brigham
            MD, Sanjana NE, Zhang F
  TITLE     Genome-scale CRISPR-Cas9 knockout and transcriptional activation 
            screening.
  JOURNAL   Nat Protoc. 2017 Apr;12(4):828-863. doi: 10.1038/nprot.2017.016. 
            Epub 2017 Mar 23.
  PUBMED    28333914
REFERENCE   2  (bases 1 to 940)
  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"; doi: 
            "10.1038/nprot.2017.016"; journalName: "Nat Protoc"; date: 
            "2017-04"; volume: "12"; issue: "4"; pages: "828-863"
COMMENT     Sequence Label: lentiSAMv2
FEATURES             Location/Qualifiers
     source          1..940
                     /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..515)
                     /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"
     CDS             complement(537..686)
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             complement(711..731)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
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 ttgggccagg attctcctcg
      481 acgtcaccgc atgttagcag acttcctctg ccctctccac tgcctgtaca gttaatcagc
      541 atgtccaggt cgaaatcatc aagggcgtca ctgccgagca tgtcgaggtc aaagtcatca
      601 agggcatccg aaccaagcat gtcaaggtca aaatcatcga gggcgtcact tcccagcata
      661 tccagatcaa aatcgtccaa tgcgtcagcc cgtccggatc cagcggccgc caccttcctc
      721 tttttcttag gtccgctacc tcctcctccg cttcctcctc ctccgcttcc acctcctcca
      781 gcgctgtcgc ctcccagctg agacaggtcg atccgtgtct cgtacaggcc ggtgatgctc
      841 tggtggatca gggtggcgtc cagcacctct ttggtgctgg tgtacctctt ccggtcgatg
      901 gtggtgtcaa agtacttgaa ggcggcaggg gctcccagat
//
