LOCUS       sequence-162017-         998 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  The sgRNA at the stop codon of human Brachyury(T) locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 998)
  AUTHORS   Chu LF, Leng N, Zhang J, Hou Z, Mamott D, Vereide DT, Choi J, 
            Kendziorski C, Stewart R, Thomson JA
  TITLE     Single-cell RNA-seq reveals novel regulators of human embryonic stem
            cell differentiation to definitive endoderm.
  JOURNAL   Genome Biol. 2016 Aug 17;17(1):173. doi: 10.1186/s13059-016-1033-x.
  PUBMED    27534536
REFERENCE   2  (bases 1 to 998)
  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: "Genome 
            Biol."; date: "2016-08-17"; volume: "17(1):173. doi"; pages: " 
            10.1186/s13059-016-1033-x"
COMMENT     Sequence Label: T-STOP sgRNA-1
FEATURES             Location/Qualifiers
     source          1..998
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     216..233
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    350..371
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(386..391,392..409,410..416)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    424..440
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     429..451
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     448..464
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     448..464
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        482..500
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     482..499
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        649..897
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     649..669
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     820..839
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        918..993
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
       61 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      121 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      181 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      241 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
      301 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
      361 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
      421 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
      481 tatttaggtg acactataga atactcaagc tatgcatcaa gcttggtacc gagctcggat
      541 ccactagtaa cggccgccag tgtgctggaa ttcgcccttt gtacaaaaaa gcaggcttta
      601 aaggaaccaa ttcagtcgac tggatccggt accaaggtcg ggcaggaaga gggcctattt
      661 cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat aattagaatt
      721 aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt
      781 cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta
      841 acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccgcc
      901 ttccatgtga agcagcagtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
      961 tatcaacttg aaaaagtggc accgagtcgg tgcttttt
//
