LOCUS       sequence-107707-        3012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses sgRNA under U6c promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3012)
  AUTHORS   Yin L, Maddison LA, Li M, Kara N, LaFave MC, Varshney GK, Burgess 
            SM, Patton JG, Chen W
  TITLE     Multiplex Conditional Mutagenesis Using Transgenic Expression of 
            Cas9 and sgRNAs.
  JOURNAL   Genetics. 2015 Apr 8. pii: genetics.115.176917.
  PUBMED    25855067
REFERENCE   2  (bases 1 to 3012)
  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: "Genetics. 
            2015 Apr 8. pii: genetics.115.176917."
COMMENT     Sequence Label: pU6c:sgRNA#4
FEATURES             Location/Qualifiers
     source          1..3012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             942..1733
                     /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"
     rep_origin      1826..2414
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2315..2334
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2568..2585
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2744..2771)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2863..2949)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(2998..3012,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3012,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc ccggtctctc caatccgaga gtctgtgaat
       61 gtttcgatgg gccgtttgct ggcgtctgag catgttgtcc tctggtctgc ggcgggggaa
      121 ggtccgcgca tgcctgtggg ggaaggcgaa ggacacgagg acaaaagatc agttacgtca
      181 cagaagaact catccaatca cacaagccta aaaccagctt cctctgcata tgctttacag
      241 ttcaaaaata ctacggtaaa cctctacaaa actgctggtt tcaaactttg aaggttttaa
      301 gcgtttgcag gtttgccccg aagaggttta ctgtcatgtt tgaggtaaat gagttgaata
      361 agtaggttta tccacttacc acatggcaga aacataccca gaagtcccgg gtttatatag
      421 cagttctcca ggctcctagt tcgagagacg acactgcaga ctgtcgaccg tctctgttta
      481 agagctatgc tggaaacagc atagcaagtt taaataaggc tagtccgtta tcaacttgaa
      541 aaagtggcac cgagtcggtg ctttttttaa acctgagaga cctctagacc agccaggaca
      601 gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg acgcacaccg tggaaacgga
      661 tgaaggcacg aacccagtgg acataagcct gttcggttcg taagctgtaa tgcaagtagc
      721 gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag
      781 tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat
      841 ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg
      901 ttacgcagca gggcagtcgc cctaaaacaa agttaaacat tatgagggaa gcggtgatcg
      961 ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga
     1021 cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg
     1081 atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga
     1141 tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag
     1201 aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac
     1261 tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga
     1321 tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag
     1381 gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa
     1441 atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag
     1501 tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg
     1561 tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag
     1621 ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg
     1681 aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga
     1741 gccacccatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
     1801 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     1861 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     1921 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     1981 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2041 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2101 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2161 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagcatt
     2221 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2281 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2341 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2401 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2461 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2521 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2581 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     2641 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     2701 ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag
     2761 gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc
     2821 tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag
     2881 agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg
     2941 ttttatttga tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc
     3001 agtcacgacg tt
//
