LOCUS       sequence-158326-        2191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pHu6-gRNA-NT1_SpecR backbone, gRNA for targeting the 5' region of 
            pCALNL_Ch13 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_Ch13_102010574-gRNA-rev
FEATURES             Location/Qualifiers
     source          1..2191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(454..558)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        659..907
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     659..679
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     830..849
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        929..1004
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(1097..1685)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1177..1196)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1856..2191,1..453))
                     /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 ctccgccgct ggacctacca aggcaacgct atgttctctt gcttttgtca gcaagatagc
       61 cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat tgcgctgcca
      121 ttctccaaat tgcagttcgc gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac
      181 aacaatggtg acttctacag cgcggagaat ctcgctctct ccaggggaag ccgaagtttc
      241 caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac
      301 cagcaaatca atatcactgt gtggcttcag gccgccatcc actgcggagc cgtacaaatg
      361 tacggccagc aacgtcggtt cgagatggcg ctcgatgacg ccaactacct ctgatagttg
      421 agtcgatact tcggcgatca ccgcttccct catactcttc ctttttcaat attattgaag
      481 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
      541 acaaatatgg gttccgcgca catttccccg aaaagtgcca cctgacgtcg ctagctgtac
      601 aaaaaagcag gctttaaagg aaccaattca ttcgactgga tccaaggtcg ggcaggaaga
      661 gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat
      721 aattagaatt aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa
      781 gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat
      841 gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg
      901 aaacaccgtc agctatgatc acttccctgt tttagagcta gaaatagcaa gttaaaataa
      961 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt ttctcgaggt
     1021 acctctctac atatgacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1081 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1141 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1201 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1261 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1321 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1381 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1441 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1501 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     1561 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1621 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1681 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1741 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1801 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagtttgc
     1861 cgactacctt ggtgatctcg cctttcacgt agtggacaaa ttcttccaac tgatctgcgc
     1921 gcgaggccaa gcgatcttct tcttgtccaa gataagcctg tctagcttca agtatgacgg
     1981 gctgatactg ggccggcagg cgctccattg cccagtcggc agcgacatcc ttcggcgcga
     2041 ttttgccggt tactgcgctg taccaaatgc gggacaacgt aagcactaca tttcgctcat
     2101 cgccagccca gtcgggcggc gagttccata gcgttaaggt ttcatttagc gcctcaaata
     2161 gatcctgttc aggaaccgga tcaaagagtt c
//
