LOCUS       sequence-158313-        2262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for targeting Ch10 psuedo gix site in PCDH15 gene (five 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-for2
FEATURES             Location/Qualifiers
     source          1..2262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     552..571
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(790..894)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        995..1243
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     995..1015
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1166..1185
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        1264..1339
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(1432..2020)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1512..1531)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2191..2262,1..720,721..789))
                     /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 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
       61 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
      121 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
      181 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
      241 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
      301 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
      361 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
      421 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
      481 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
      541 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
      601 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
      661 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
      721 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
      781 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
      841 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
      901 tccccgaaaa gtgccacctg acgtcgctag ctgtacaaaa aagcaggctt taaaggaacc
      961 aattcagtcg actggatcca aggtcgggca ggaagagggc ctatttccca tgattccttc
     1021 atatttgcat atacgataca aggctgttag agagataatt agaattaatt tgactgtaaa
     1081 cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc
     1141 agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt
     1201 cgatttcttg gctttatata tcttgtggaa aggacgaaac accggcccat gacccttctc
     1261 ctcgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
     1321 agtggcaccg agtcggtgct tttttttctc gaggtacctc tctacatatg acatgtgagc
     1381 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1441 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1501 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1561 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1621 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1681 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1741 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1801 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1861 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1921 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1981 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2041 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2101 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2161 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2221 ctcagcgatc tgtctatttc gttcatccat agttgcctga ct
//
