LOCUS       sequence-107705-        3014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses sgRNA under U6a promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3014)
  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 3014)
  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: pU6a:sgRNA#2
FEATURES             Location/Qualifiers
     source          1..3014
                     /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             944..1735
                     /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      1828..2416
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2317..2336
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2570..2587
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2746..2773)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2865..2951)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3000..3014,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3014,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc ccggtctcta gtctttctcc agcctcggtc
       61 attcagtaag acgagtttcc tgctctcggc gtcttttgtt ctggtcttca aggagggggg
      121 aaatgttctg cgcatgcctg tgggggaggg agaaggacac gtcactgaaa acgtccctgc
      181 atcacaccga gacacccaat cactcaagcc aagaccagat aattttgcat atgctttaca
      241 gtttgaaaat taccacggta atccctcaca caaactctgg atttgagatc tttcaggttt
      301 tatcagtttg caggtttatg tcaccatgat atagggtcag acttgatcta agggagctga
      361 ataagtggtt tagtcactca ccacctccca aaaacatacc cagaagtccc tggtatatat
      421 agctctccct ccagctcttg gttcgagaga cgacactgca gactgtcgac cgtctctgtt
      481 taagagctat gctggaaaca gcatagcaag tttaaataag gctagtccgt tatcaacttg
      541 aaaaagtggc accgagtcgg tgcttttttt aaactcaaga gacctctaga ccagccagga
      601 cagaaatgcc tcgacttcgc tgctacccaa ggttgccggg tgacgcacac cgtggaaacg
      661 gatgaaggca cgaacccagt ggacataagc ctgttcggtt cgtaagctgt aatgcaagta
      721 gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc
      781 agtggcggtt ttcatggctt gttatgactg tttttttggg gtacagtcta tgcctcgggc
      841 atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga
      901 tgttacgcag cagggcagtc gccctaaaac aaagttaaac attatgaggg aagcggtgat
      961 cgccgaagta tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc
     1021 gacgttgctg gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag
     1081 tgatattgat ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt
     1141 gatcaacgac cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt
     1201 agaagtcacc attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga
     1261 actgcaattt ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac
     1321 gatcgacatt gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt
     1381 aggtccagcg gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct
     1441 aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt
     1501 agtgcttacg ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga
     1561 tgtcgctgcc gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga
     1621 agctagacag gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt
     1681 ggaagaattt gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataaccctc
     1741 gagccaccca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     1801 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1861 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1921 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     1981 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2041 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2101 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2161 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca
     2221 ttgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2281 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2341 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2401 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2461 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     2521 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     2581 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     2641 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     2701 aattaatacg cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc
     2761 aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac
     2821 cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg
     2881 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt
     2941 cgttttattt gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc
     3001 ccagtcacga cgtt
//
