LOCUS       sequence-113695-         960 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  3rd generation lentiviral vector that expresses shRNA under the 
            mouse U6 promoter. A CMV-EGFP reporter cassette is included in the 
            vector to monitor expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 960)
  AUTHORS   Rubinson DA, Dillon CP, Kwiatkowski AV, Sievers C, Yang L, Kopinja 
            J, Rooney DL, Ihrig MM, McManus MT, Gertler FB, Scott ML, Van Parijs
            L
  TITLE     A lentivirus-based system to functionally silence genes in primary 
            mammalian cells, stem cells and transgenic mice by RNA interference.
  JOURNAL   Nat Genet. 2003 Mar . 33(3):401-6.
  PUBMED    12590264
REFERENCE   2  (bases 1 to 960)
  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: "Nat Genet. 
            2003 Mar . 33(3):401-6."
COMMENT     Sequence Label: pLL3.7
FEATURES             Location/Qualifiers
     source          1..960
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    81..114
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(134..847,848..850,851..853))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(137..847,848..850,851..853))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(179..200)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     528..547
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     787..808
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      850..859
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(898..949)
                     /label=CMVd2 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields moderately reduced expression 
                     relative to the full-length promoter"
     primer_bind     complement(927..947)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 aaattttgta atccagaggt tgattatcga taagcttgat atcgaattgt acctagtgga
       61 accggaaccc ttaaacatgt ataacttcgt ataatgtatg ctatacgaag ttattaggtc
      121 cctcgacgaa ttcctacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
      181 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact
      241 gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
      301 gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt
      361 tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt
      421 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
      481 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
      541 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
      601 agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca
      661 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca
      721 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
      781 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
      841 ccttgctcac catggtggcg accggtagcg ctagcggatc tgacggttca ctaaaccagc
      901 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga
//
