LOCUS       sequence-258912-        1009 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  gRNA used for knockin NanoLuc and tdTomado (separated by 2A) into 
            MYH11 allele (MYH11-NanoLuc-2A-tdTomato vector) ).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1009)
  AUTHORS   Zhang J, McIntosh BE, Wang B, Brown ME, Probasco MD, Webster S, 
            Duffin B, Zhou Y, Guo LW, Burlingham WJ, Kent C, Ferris M, Thomson 
            JA
  TITLE     A Human Pluripotent Stem Cell-Based Screen for Smooth Muscle Cell 
            Differentiation and Maturation Identifies Inhibitors of Intimal 
            Hyperplasia.
  JOURNAL   Stem Cell Reports. 2019 Jun 11;12(6):1269-1281. doi: 
            10.1016/j.stemcr.2019.04.013. Epub 2019 May 9.
  PUBMED    31080110
REFERENCE   2  (bases 1 to 1009)
  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.stemcr.2019.04.013"; journalName: "Stem Cell Reports"; 
            date: "2019-06-11- 11"; volume: "12"; issue: "6"; pages: "1269-1281"
COMMENT     Sequence Label: MYH11-gRNA1
FEATURES             Location/Qualifiers
     source          1..1009
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        88..336
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     88..108
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     259..278
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        357..432
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(543..561)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(544..561)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(575..592)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(575..591)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(584..606)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(869..890)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cggcgcagct gcngngcgtt ncgttatcgg atccagaatt cgtgatccaa gctcggatcc
       61 gatatcgacg tcaaggtcgg gcaggaagag ggcctatttc ccatgattcc ttcatatttg
      121 catatacgat acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag
      181 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
      241 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
      301 ttggctttat atatcttgtg gaaaggacga aacaccgaag gcaccatggc gcagaagttt
      361 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
      421 ccgagtcggt gctttttttt gcggccgcac ttgaaattcg atctgaattc gtcgacaagc
      481 ttctcgagcc taggctagct ctagaccaca cgtgtggggg cccgagctcg cggccgctgt
      541 attctatagt gtcacctaaa tggccgcaca attcactggc cgtcgtttta caacgtcgtg
      601 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
      661 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
      721 atggcgaatg gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
      781 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
      841 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
      901 cgtggactcc aacgtcaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      961 ccatcaccct aatcaagntt ttttnggggt cgaggtgccg taaagcact
//
