LOCUS       sequence-158314-        2262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for targeting Ch10 psuedo gix site in PCDH15 gene(six base pairs 
            from the cas9 site and 3' of gix site).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2262)
  AUTHORS   Chaikind B, Bessen JL, Thompson DB, Hu JH, Liu DR
  TITLE     A programmable Cas9-serine recombinase fusion protein that operates 
            on DNA sequences in mammalian cells.
  JOURNAL   Nucleic Acids Res. 2016 Aug 11. pii: gkw707.
  PUBMED    27515511
REFERENCE   2  (bases 1 to 2262)
  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: "Nucleic 
            Acids Res. 2016 Aug 11. pii: gkw707."
COMMENT     Sequence Label: pHU6_gRNA_Ch10-for1
FEATURES             Location/Qualifiers
     source          1..2262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     524..543
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(762..866)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        967..1215
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     967..987
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1138..1157
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        1236..1311
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(1404..1992)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1484..1503)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2163..2262,1..692,693..761))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
       61 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
      121 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
      181 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
      241 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
      301 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
      361 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
      421 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
      481 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
      541 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
      601 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
      661 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
      721 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
      781 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
      841 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgct
      901 agctgtacaa aaaagcaggc tttaaaggaa ccaattcagt cgactggatc caaggtcggg
      961 caggaagagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     1021 agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     1081 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     1141 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     1201 aaaggacgaa acaccgccca tgacccttct cctctgtttt agagctagaa atagcaagtt
     1261 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttttc
     1321 tcgaggtacc tctctacata tgacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1381 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1441 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1501 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1561 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1621 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1681 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1741 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1801 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1861 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1921 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1981 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2041 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2101 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2161 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2221 atagttgcct gactccccgt cgtgtagata actacgatac gg
//
