LOCUS       sequence-167859-         802 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  hU6 driven sgRNA vector with 2xMS2 vectors with puro selectable 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 802)
  AUTHORS   Hess GT, Fresard L, Han K, Lee CH, Li A, Cimprich KA, Montgomery SB,
            Bassik MC
  TITLE     Directed evolution using dCas9-targeted somatic hypermutation in 
            mammalian cells.
  JOURNAL   Nat Methods. 2016 Oct 31. doi: 10.1038/nmeth.4038.
  PUBMED    27798611
REFERENCE   2  (bases 1 to 802)
  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: "Nat 
            Methods. 2016 Oct 31. doi: 10.1038/nmeth.4038."
COMMENT     Sequence Label: pGH224_sgRNA_2xMS2_Puro
FEATURES             Location/Qualifiers
     source          1..802
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     125..144
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        240..258
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        310..328
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     promoter        471..682
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
ORIGIN
        1 ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa
       61 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa
      121 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat
      181 cttgtggaaa ggacgaaaca ccggagacgg gataccgtct ctgttttaga gctaggccaa
      241 catgaggatc acccatgtct gcagggccta gcaagttaaa ataaggctag tccgttatca
      301 acttggccaa catgaggatc acccatgtct gcagggccaa gtggcaccga gtcggtgctt
      361 tttttggatc ctgcaaagat ggataaagtt ttaaacagag aggaatcttt gcagctaatg
      421 gaccttctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg
      481 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta
      541 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
      601 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
      661 cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt
      721 tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt acgtgattct
      781 tgatcccgag cttcgggttg ga
//
