LOCUS       sequence-163283-        2191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pHu6-gRNA-NT1_SpecR backbone, gRNA for targeting the upstream region
            of FAM19A2 locus.
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_chr12_FAM19A2-up-gRNA-rev
FEATURES             Location/Qualifiers
     source          1..2191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        76..324
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     76..96
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     247..266
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        346..421
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(514..1102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(594..613)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1273..2061)
                     /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"
     promoter        complement(2062..2166)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcattc gactggatcc
       61 aaggtcgggc aggaagaggg cctatttccc atgattcctt catatttgca tatacgatac
      121 aaggctgtta gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa
      181 aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt
      241 aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat
      301 atcttgtgga aaggacgaaa caccgatatc cgtttatcag tgtcagtttt agagctagaa
      361 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
      421 cttttttttc tcgaggtacc tctctacata tgacatgtga gcaaaaggcc agcaaaaggc
      481 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
      541 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
      601 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
      661 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
      721 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
      781 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
      841 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
      901 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
      961 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     1021 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     1081 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     1141 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     1201 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     1261 ttggtctgac agtttgccga ctaccttggt gatctcgcct ttcacgtagt ggacaaattc
     1321 ttccaactga tctgcgcgcg aggccaagcg atcttcttct tgtccaagat aagcctgtct
     1381 agcttcaagt atgacgggct gatactgggc cggcaggcgc tccattgccc agtcggcagc
     1441 gacatccttc ggcgcgattt tgccggttac tgcgctgtac caaatgcggg acaacgtaag
     1501 cactacattt cgctcatcgc cagcccagtc gggcggcgag ttccatagcg ttaaggtttc
     1561 atttagcgcc tcaaatagat cctgttcagg aaccggatca aagagttcct ccgccgctgg
     1621 acctaccaag gcaacgctat gttctcttgc ttttgtcagc aagatagcca gatcaatgtc
     1681 gatcgtggct ggctcgaaga tacctgcaag aatgtcattg cgctgccatt ctccaaattg
     1741 cagttcgcgc ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa caatggtgac
     1801 ttctacagcg cggagaatct cgctctctcc aggggaagcc gaagtttcca aaaggtcgtt
     1861 gatcaaagct cgccgcgttg tttcatcaag ccttacggtc accgtaacca gcaaatcaat
     1921 atcactgtgt ggcttcaggc cgccatccac tgcggagccg tacaaatgta cggccagcaa
     1981 cgtcggttcg agatggcgct cgatgacgcc aactacctct gatagttgag tcgatacttc
     2041 ggcgatcacc gcttccctca tactcttcct ttttcaatat tattgaagca tttatcaggg
     2101 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaatatgggt
     2161 tccgcgcaca tttccccgaa aagtgccacc t
//
