LOCUS       sequence-158321-        2191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pHu6-gRNA-NT1_SpecR backbone, gRNA for targeting the 3' 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-for
FEATURES             Location/Qualifiers
     source          1..2191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(620..724)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        825..1073
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     825..845
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     996..1015
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        1095..1170
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(1263..1851)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1343..1362)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2022..2191,1..619))
                     /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 catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac aacgtaagca
       61 ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat
      121 ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc gccgctggac
      181 ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga
      241 tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct ccaaattgca
      301 gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt
      361 ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa aggtcgttga
      421 tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat
      481 cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg gccagcaacg
      541 tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc gatacttcgg
      601 cgatcaccgc ttccctcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
      661 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa atatgggttc
      721 cgcgcacatt tccccgaaaa gtgccacctg acgtcgctag ctgtacaaaa aagcaggctt
      781 taaaggaacc aattcattcg actggatcca aggtcgggca ggaagagggc ctatttccca
      841 tgattccttc atatttgcat atacgataca aggctgttag agagataatt agaattaatt
      901 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg
      961 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
     1021 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac accgaaatag
     1081 cacaatgaat ggaagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat
     1141 caacttgaaa aagtggcacc gagtcggtgc ttttttttct cgaggtacct ctctacatat
     1201 gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1261 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     1321 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     1381 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     1441 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     1501 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     1561 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     1621 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     1681 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     1741 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     1801 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     1861 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     1921 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     1981 aaatcaatct aaagtatata tgagtaaact tggtctgaca gtttgccgac taccttggtg
     2041 atctcgcctt tcacgtagtg gacaaattct tccaactgat ctgcgcgcga ggccaagcga
     2101 tcttcttctt gtccaagata agcctgtcta gcttcaagta tgacgggctg atactgggcc
     2161 ggcaggcgct ccattgccca gtcggcagcg a
//
