LOCUS       sequence-138993-         983 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  3rd generation lenti sgRNA cloning backbone with MS2 loops at 
            tetraloop and stemloop 2 and EF1a-puro resistance marker. Contains 
            BsmBI sites for insertion of spacer sequences..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 983)
  AUTHORS   Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena 
            C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F
  TITLE     Genome-scale transcriptional activation by an engineered CRISPR-Cas9
            complex.
  JOURNAL   Nature. 2014 Dec 10. doi: 10.1038/nature14136.
  PUBMED    25494202
REFERENCE   2  (bases 1 to 983)
  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: "Nature. 
            2014 Dec 10. doi: 10.1038/nature14136."
COMMENT     Sequence Label: lenti sgRNA(MS2)_puro backbone
FEATURES             Location/Qualifiers
     source          1..983
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(63..662)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(146..166)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     643..662
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     intron          678..881
                     /label=EF-1-alpha intron A
                     /note="truncated intron upstream of the start codon of 
                     human EF-1-alpha"
     primer_bind     complement(701..721)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 gtcaatcttt cacaaatttt gtaatccaga ggttgattac cgataagctt gatatcgaat
       61 tctcaggcac cgggcttgcg ggtcatgcac caggtgcgcg gtccttcggg cacctcgacg
      121 tcggcggtga cggtgaagcc gagccgctcg tagaagggga ggttgcgggg cgcggaggtc
      181 tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca ctccggggag cacgacggcg
      241 ctgcccagac ccttgccctg gtggtcgggc gagactccga cggtggccag gaaccacgcg
      301 ggctccttgg gccggtgcgg cgccaggagg ccttccatct gttgctgcgc ggccagccgg
      361 gaaccgctca actcggccat gcgcgggccg atctcggcga acaccgcccc cgcttcgacg
      421 ctctccggcg tggtccagac cgccacggcg gcgccgtcgt ccgcgaccca caccttgccg
      481 atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg ctcgatgtgg
      541 cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc ggcgagggtg
      601 cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt gtactcggtc
      661 attgtacatc acgacacctg aaatggaaga aaaaaacttt gaaccactgt ctgaggcttg
      721 agaatgaacc aagatccaaa ctcaaaaagg gcaaattcca aggagaatta catcaagtgc
      781 caagctggcc taacttcagt ctccacccac tcagtgtggg gaaactccat cgcataaaac
      841 ccctcccccc aacctaaaga cgacgtactc caaaagctcg agaactaatc gaggtgcctg
      901 gacggcgccc ggtactccgt ggagtcacat gaagcgacgg ctgaggacgg aaaggccctt
      961 ttcctttgtg tgggtgactc acc
//
