LOCUS       sequence-158324-        2190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pHu6-gRNA-NT1_SpecR backbone, gRNA for targeting the 3' region of 
            pCALNL_Ch13 reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2190)
  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 2190)
  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_Ch13_102010574-gRNA-for
FEATURES             Location/Qualifiers
     source          1..2190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(771..875)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        976..1224
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     976..996
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1147..1166
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        1245..1320
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(1413..2001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1493..1512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2172..2190,1..770))
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 atctcgcctt tcacgtagtg gacaaattct tccaactgat ctgcgcgcga ggccaagcga
       61 tcttcttctt gtccaagata agcctgtcta gcttcaagta tgacgggctg atactgggcc
      121 ggcaggcgct ccattgccca gtcggcagcg acatccttcg gcgcgatttt gccggttact
      181 gcgctgtacc aaatgcggga caacgtaagc actacatttc gctcatcgcc agcccagtcg
      241 ggcggcgagt tccatagcgt taaggtttca tttagcgcct caaatagatc ctgttcagga
      301 accggatcaa agagttcctc cgccgctgga cctaccaagg caacgctatg ttctcttgct
      361 tttgtcagca agatagccag atcaatgtcg atcgtggctg gctcgaagat acctgcaaga
      421 atgtcattgc gctgccattc tccaaattgc agttcgcgct tagctggata acgccacgga
      481 atgatgtcgt cgtgcacaac aatggtgact tctacagcgc ggagaatctc gctctctcca
      541 ggggaagccg aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc
      601 cttacggtca ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc gccatccact
      661 gcggagccgt acaaatgtac ggccagcaac gtcggttcga gatggcgctc gatgacgcca
      721 actacctctg atagttgagt cgatacttcg gcgatcaccg cttccctcat actcttcctt
      781 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
      841 tgtatttaga aaaataaaca aatatgggtt ccgcgcacat ttccccgaaa agtgccacct
      901 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcattc gactggatcc
      961 aaggtcgggc aggaagaggg cctatttccc atgattcctt catatttgca tatacgatac
     1021 aaggctgtta gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa
     1081 aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt
     1141 aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat
     1201 atcttgtgga aaggacgaaa caccggagac ttaagtccaa aaccgtttta gagctagaaa
     1261 tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
     1321 ttttttttct cgaggtacct ctctacatat gacatgtgag caaaaggcca gcaaaaggcc
     1381 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1441 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1501 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1561 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1621 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1681 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1741 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1801 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1861 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1921 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1981 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2041 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2101 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2161 tggtctgaca gtttgccgac taccttggtg
//
