LOCUS       Exported                3046 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  expresses sgRNA under U6b promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pU6b:sgRNA#3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3046)
  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 3046)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3046
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             976..1767
                     /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      1860..2448
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2349..2368
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2602..2619
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      2778..2805
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      2897..2983
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3032..8
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3046..17
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc ccggtctctc tcattaccct ccacgtgtct
       61 gtctgggttt tcatgggctc tgctctagtg agagcagctt ccttttgtcg gagtgttttg
      121 tgctcttatc caaggcaggg gggatctgcg catgcctgtg gggggaggag aaggacacgt
      181 gaacaaaagc tcctcgatgt cacacaggaa gttcaggaac tcatccaatc actctaaaga
      241 aacggcctgt ttccttcgca tacgcttaca gctccaaaac tctacggtaa acctacataa
      301 actgctggtt ttcaaatttt aaagaattta agggtttaca ggtttactac tacacagtga
      361 tttactgaca catgtaagtg taaatgagtt gaataagtaa gtaagccata taccacacat
      421 gaaacacata cccagaagtc actggtatat atagccgtcc tccagactcc cagttcgaga
      481 gacgacactg cagactgtcg accgtctctg tttaagagct atgctggaaa cagcatagca
      541 agtttaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
      601 ttaaaccaaa gagacctcta gaccagccag gacagaaatg cctcgacttc gctgctaccc
      661 aaggttgccg ggtgacgcac accgtggaaa cggatgaagg cacgaaccca gtggacataa
      721 gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga
      781 accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac
      841 tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg
      901 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa
      961 acaaagttaa acattatgag ggaagcggtg atcgccgaag tatcgactca actatcagag
     1021 gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc
     1081 tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc
     1141 gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct
     1201 tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac
     1261 atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat
     1321 gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg
     1381 acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat
     1441 ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg
     1501 ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac
     1561 agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc
     1621 ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa
     1681 gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc
     1741 gagatcacca aggtagtcgg caaataaccc tcgagccacc catgaccaaa atcccttaac
     1801 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1861 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1921 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1981 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     2041 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2101 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2161 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2221 tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg
     2281 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2341 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2401 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2461 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2521 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2581 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     2641 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     2701 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     2761 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     2821 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     2881 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     2941 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     3001 actctcgcgt taacgctagc atggatgttt tcccagtcac gacgtt
//
