LOCUS       sequence-158320-        2191 bp    DNA     circular SYN 15-APR-2025
DEFINITION  pHu6-gRNA-NT1_SpecR backbone, gRNA for targeting the 5' region of 
            pCALNL_Ch5 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 Apr 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-rev
FEATURES             Location/Qualifiers
     source          1..2191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(509..613)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        714..962
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     714..734
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     885..904
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        984..1059
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(1152..1740)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1232..1251)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1911..2191,1..508))
                     /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 aggtttcatt tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg
       61 ccgctggacc taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat
      121 caatgtcgat cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc tgccattctc
      181 caaattgcag ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa
      241 tggtgacttc tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa
      301 ggtcgttgat caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaaccagca
      361 aatcaatatc actgtgtggc ttcaggccgc catccactgc ggagccgtac aaatgtacgg
      421 ccagcaacgt cggttcgaga tggcgctcga tgacgccaac tacctctgat agttgagtcg
      481 atacttcggc gatcaccgct tccctcatac tcttcctttt tcaatattat tgaagcattt
      541 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
      601 tatgggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgctagc tgtacaaaaa
      661 agcaggcttt aaaggaacca attcattcga ctggatccaa ggtcgggcag gaagagggcc
      721 tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga gagataatta
      781 gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat
      841 aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta
      901 ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca
      961 ccgtctacct ctcatgagcc ggtgttttag agctagaaat agcaagttaa aataaggcta
     1021 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttttctc gaggtacctc
     1081 tctacatatg acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1141 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1201 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1261 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1321 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1381 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1441 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1501 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1561 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     1621 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1681 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1741 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     1801 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     1861 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag tttgccgact
     1921 accttggtga tctcgccttt cacgtagtgg acaaattctt ccaactgatc tgcgcgcgag
     1981 gccaagcgat cttcttcttg tccaagataa gcctgtctag cttcaagtat gacgggctga
     2041 tactgggccg gcaggcgctc cattgcccag tcggcagcga catccttcgg cgcgattttg
     2101 ccggttactg cgctgtacca aatgcgggac aacgtaagca ctacatttcg ctcatcgcca
     2161 gcccagtcgg gcggcgagtt ccatagcgtt a
//
