LOCUS       Exported                 707 bp ds-DNA     linear   SYN 18-MAY-2021
DEFINITION  Expresses Cas9 nuclease and the PITCh-gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    pX330S-2-PITCh
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 707)
  AUTHORS   Sakuma T, Nakade S, Sakane Y, Suzuki KT, Yamamoto T
  TITLE     MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the 
            PITCh systems.
  JOURNAL   Nat Protoc. 2016 Jan;11(1):118-33. doi: 10.1038/nprot.2015.140. Epub
            2015 Dec 17.
  PUBMED    26678082
REFERENCE   2  (bases 1 to 707)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 18, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..707
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     117..136
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        215..290
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        386..671
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
       61 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
      121 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
      181 aaggacgaaa caccgcatcg tacgcgtacg tgttgtttta gagctagaaa tagcaagtta
      241 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttgttt
      301 tagagctaga aatagcaagt taaaataagg ctagtccgtt tttagcgcgt gcgccaattc
      361 tgcagacaaa tggctctatg agacccgtta cataacttac ggtaaatggc ccgcctggct
      421 gaccgcccaa cgacccccgc ccattgacgt caatagtaac gccaataggg actttccatt
      481 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      541 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattgtg
      601 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      661 ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctcccc
//
