LOCUS       sequence-158323-        2191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pHu6-gRNA-NT1_SpecR backbone, gRNA for targeting the 5' region of 
            pCALNL_Ch12 reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2191)
  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 2191)
  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_Ch12_62418577-gRNA-rev
FEATURES             Location/Qualifiers
     source          1..2191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(488..592)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        693..941
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     693..713
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     864..883
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        963..1038
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(1131..1719)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1211..1230)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1890..2191,1..487))
                     /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 atagatcctg ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa
       61 cgctatgttc tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct
      121 cgaagatacc tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag
      181 ctggataacg ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga
      241 gaatctcgct ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc
      301 gcgttgtttc atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct
      361 tcaggccgcc atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat
      421 ggcgctcgat gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt
      481 ccctcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
      541 gcggatacat atttgaatgt atttagaaaa ataaacaaat atgggttccg cgcacatttc
      601 cccgaaaagt gccacctgac gtcgctagct gtacaaaaaa gcaggcttta aaggaaccaa
      661 ttcattcgac tggatccaag gtcgggcagg aagagggcct atttcccatg attccttcat
      721 atttgcatat acgatacaag gctgttagag agataattag aattaatttg actgtaaaca
      781 caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag
      841 ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg
      901 atttcttggc tttatatatc ttgtggaaag gacgaaacac cgactttttg ggggagaggg
      961 aggttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa
     1021 gtggcaccga gtcggtgctt ttttttctcg aggtacctct ctacatatga catgtgagca
     1081 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1141 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1201 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1261 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1321 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1381 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1441 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1501 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1561 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1621 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1681 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1741 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     1801 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     1861 agtatatatg agtaaacttg gtctgacagt ttgccgacta ccttggtgat ctcgcctttc
     1921 acgtagtgga caaattcttc caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt
     1981 ccaagataag cctgtctagc ttcaagtatg acgggctgat actgggccgg caggcgctcc
     2041 attgcccagt cggcagcgac atccttcggc gcgattttgc cggttactgc gctgtaccaa
     2101 atgcgggaca acgtaagcac tacatttcgc tcatcgccag cccagtcggg cggcgagttc
     2161 catagcgtta aggtttcatt tagcgcctca a
//
