LOCUS       sequence-160226-         971 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  Expresses gRNA of interest under U6 promoter (cloning by BpiI) and 
            GFP under PGK promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 971)
  AUTHORS   Daneshvar K, Pondick JV, Kim BM, Zhou C, York SR, Macklin JA, 
            Abualteen A, Tan B, Sigova AA, Marcho C, Tremblay KD, Mager J, Choi 
            MY, Mullen AC
  TITLE     DIGIT Is a Conserved Long Noncoding RNA that Regulates GSC 
            Expression to Control Definitive Endoderm Differentiation of 
            Embryonic Stem Cells.
  JOURNAL   Cell Rep. 2016 Oct 4;17(2):353-365. doi: 
            10.1016/j.celrep.2016.09.017.
  PUBMED    27705785
REFERENCE   2  (bases 1 to 971)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2016.09"; journalName: "Cell Rep"; date: 
            "2016-10-4- 4"; volume: "17"; issue: "2"; pages: "353-365"
COMMENT     Sequence Label: pgRGFP
FEATURES             Location/Qualifiers
     source          1..971
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        124..372
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     124..144
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     295..314
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        391..466
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(608..631)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
       61 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      121 tgtgagggcc tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga
      181 gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt
      241 agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat
      301 catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa
      361 ggacgaaaca ccgggtcttc gagaagacct gttttagagc tagaaatagc aagttaaaat
      421 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
      481 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
      541 gacaaatggc tcgtcgacct cgaaattcta ccgggtaggg gaggcgcttt tcccaaggca
      601 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg
      661 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc
      721 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc
      781 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag
      841 caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc
      901 cttcgctttc tgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg
      961 gaggcccggc a
//
