LOCUS       sequence-205603-        3023 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 3023)
  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 3023)
  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#1
FEATURES             Location/Qualifiers
     source          1..3023
                     /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             953..1744
                     /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      1837..2425
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2326..2345
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2579..2596
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2755..2782)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2874..2960)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3009..3023,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3023,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc ccggtctctc agtaagtctc tcagcgtttc
       61 tccagcctcg gtcattcagt aagacgagtt tcctgctctc ggcgtctttt gttctggtct
      121 tcaaggaggg gggaaatgtt ctgcgcatgc ctgtggggga gggagaagga cacgtcactg
      181 aaaacgtccc tgcatcacac cgagacaccc aatcactcaa gccaagacca gataattttg
      241 catatgcttt acagtttgaa aattaccacg gtaatccctc acacaaactc tggatttgag
      301 atccttcagg ttttatcagt ttgcaggttt atgtcaccat gatatagggt cagacttgat
      361 ctaagggagc tgaataagtg gtttagtcac tcaccacctc ccaaaaacat acccagaagt
      421 ccctggtata tatagctctc cctccagctc ttggttcgag agacgacact gcagactgtc
      481 gaccgtctct gtttaagagc tatgctggaa acagcatagc aagtttaaat aaggctagtc
      541 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttagtcagag acctctagac
      601 cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc
      661 gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta
      721 atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg
      781 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat
      841 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
      901 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga
      961 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
     1021 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
     1081 gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
     1141 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
     1201 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
     1261 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
     1321 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
     1381 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
     1441 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
     1501 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
     1561 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
     1621 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
     1681 agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa
     1741 ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     1801 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     1861 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     1921 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     1981 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2041 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2101 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2161 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2221 gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2281 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2341 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     2401 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     2461 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     2521 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     2581 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     2641 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     2701 gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg
     2761 ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct
     2821 gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc
     2881 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac
     2941 tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg
     3001 gatgttttcc cagtcacgac gtt
//
