LOCUS       sequence-150451-        1052 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Stable expression of shRNA targeting mouse Pnky. The shRNA (and 
            EGFP) can be excised by the addition of Cre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1052)
  AUTHORS   Ramos AD, Andersen RE, Liu SJ, Nowakowski TJ, Hong SJ, Gertz CC, 
            Salinas RD, Zarabi H, Kriegstein AR, Lim DA
  TITLE     The long noncoding RNA Pnky regulates neuronal differentiation of 
            embryonic and postnatal neural stem cells.
  JOURNAL   Cell Stem Cell. 2015 Apr 2;16(4):439-47. doi: 
            10.1016/j.stem.2015.02.007. Epub 2015 Mar 19.
  PUBMED    25800779
REFERENCE   2  (bases 1 to 1052)
  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: 
            "10.1016/j.stem.2015.02.007"; journalName: "Cell Stem Cell"; date: 
            "2015-04-2- 2"; volume: "16"; issue: "4"; pages: "439-47"
COMMENT     Sequence Label: pSicoR (EGFP) shPnky-2
FEATURES             Location/Qualifiers
     source          1..1052
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    125..158
                     /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(178..891,892..894,895..897))
                     /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(181..891,892..894,895..897))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(223..244)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     572..591
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     831..852
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      894..903
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(942..1041)
                     /label=CMVd1 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields slightly reduced expression 
                     relative to the full-length promoter"
     primer_bind     complement(971..991)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt tgtaatccag
       61 aggttgatta tcgataagct tgatatcgaa ttgtacctag tggaaccgga acccttaaac
      121 atgtataact tcgtataatg tatgctatac gaagttatta ggtccctcga cgaattccta
      181 cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac
      241 catgtgatcg cgcttctcgt tggggtcttt gctcagggcg gactgggtgc tcaggtagtg
      301 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc
      361 gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg aagttcacct tgatgccgtt
      421 cttctgcttg tcggccatga tatagacgtt gtggctgttg tagttgtact ccagcttgtg
      481 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag
      541 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa
      601 gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt
      661 catgtggtcg gggtagcggc tgaagcactg cacgccgtag gtcagggtgg tcacgagggt
      721 gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta
      781 ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc
      841 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatggt
      901 ggcgaccggt agcgctagcg gatctgacgg ttcactaaac cagctctgct tatatagacc
      961 tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt
     1021 ggaaagtccc gttgattttg gtgccaaaac aa
//
