LOCUS       sequence-158319-        2190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pHu6-gRNA-NT1_SpecR backbone, gRNA for targeting the 3' region of 
            pCALNL_Ch5 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_Ch5_155183064-gRNA-for
FEATURES             Location/Qualifiers
     source          1..2190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(384..488)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        589..837
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     589..609
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     760..779
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        858..933
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(1026..1614)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1106..1125)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1785..2190,1..383))
                     /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 tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat tgcgctgcca ttctccaaat
       61 tgcagttcgc gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg
      121 acttctacag cgcggagaat ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg
      181 ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac cagcaaatca
      241 atatcactgt gtggcttcag gccgccatcc actgcggagc cgtacaaatg tacggccagc
      301 aacgtcggtt cgagatggcg ctcgatgacg ccaactacct ctgatagttg agtcgatact
      361 tcggcgatca ccgcttccct catactcttc ctttttcaat attattgaag catttatcag
      421 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaatatgg
      481 gttccgcgca catttccccg aaaagtgcca cctgacgtcg ctagctgtac aaaaaagcag
      541 gctttaaagg aaccaattca ttcgactgga tccaaggtcg ggcaggaaga gggcctattt
      601 cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat aattagaatt
      661 aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt
      721 cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta
      781 acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccgga
      841 actcatgtga ttaactggtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
      901 tatcaacttg aaaaagtggc accgagtcgg tgcttttttt tctcgaggta cctctctaca
      961 tatgacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1021 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1081 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1141 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1201 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     1261 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     1321 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     1381 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     1441 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     1501 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     1561 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     1621 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     1681 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     1741 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagtttgcc gactaccttg
     1801 gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg cgaggccaag
     1861 cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg ctgatactgg
     1921 gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat tttgccggtt
     1981 actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc gccagcccag
     2041 tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag atcctgttca
     2101 ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct atgttctctt
     2161 gcttttgtca gcaagatagc cagatcaatg
//
