LOCUS       sequence-158610-         841 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 841)
  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 841)
  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..841
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        134..382
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     134..154
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     305..324
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        403..478
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(490..514)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(589..608)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(590..608)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(615..632)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(615..631)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(624..646)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 atcaagcttg gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcgc
       61 cctttgtaca aaaaagcagg ctttaaagga accaattcag tcgactggat ccggtaccaa
      121 ggtcgggcag gaagagggcc tatttcccat gattccttca tatttgcata tacgatacaa
      181 ggctgttaga gagataatta gaattaattt gactgtaaac acaaagatat tagtacaaaa
      241 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa
      301 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat
      361 cttgtggaaa ggacgaaaca ccgccttcca tgtgaagcag cagttttaga gctagaaata
      421 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
      481 tttttctaga cccagctttc ttgtacaaag ttggcattaa agggcgaatt ctgcagatat
      541 ccatcacact ggcggccgct cgagcatgca tctagagggc ccaattcgcc ctatagtgag
      601 tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
      661 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
      721 gcccgcaccg atcgcccttc ccaacagttg cgcagcctat acgtacggca gtttaaggtt
      781 tacacctata aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt
      841 g
//
