LOCUS       sequence-136351-        3431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector; single gRNA under OsU3 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3431)
  AUTHORS   Lowder LG, Paul JW, Baltes NJ, Voytas DF, Zhang Y, Zhang D, Tang X, 
            Zheng X, Hsieh TF, Qi Y
  TITLE     A CRISPR/Cas9 toolbox for multiplexed plant genome editing and 
            transcriptional regulation.
  JOURNAL   Plant Physiol. 2015 Aug 21. pii: pp.00636.2015.
  PUBMED    26297141
REFERENCE   2  (bases 1 to 3431)
  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: "Plant 
            Physiol. 2015 Aug 21. pii: pp.00636.2015."
COMMENT     Sequence Label: pYPQ141D
FEATURES             Location/Qualifiers
     source          1..3431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..398
                     /label=OsU3 promoter
                     /note="Oryza sativa (rice) snRNA U3 promoter"
     misc_RNA        431..506
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(595..694)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(712..730)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(747..765)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(770..786)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(770..786)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1217..2008
                     /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      2101..2689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2590..2609
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2843..2860
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3019..3046)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3138..3224)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3273..3295
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3287..3304
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3288..3304
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3320..3415
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ctagataacg caggatccta aagggatctt taaacatacg aacagatcac ttaaagttct
       61 tctgaagcaa cttaaagtta tcaggcatgc atggatcttg gaggaatcag atgtgcagtc
      121 agggaccata gcacaggaca ggcgtcttct actggtgcta ccagcaaatg ctggaagccg
      181 ggaacactgg gtacgttgga aaccacgtga tgtggagtaa gataaactgt aggagaaaag
      241 catttcgtag tgggccatga agcctttcag gacatgtatt gcagtatggg ccggcccatt
      301 acgcaattgg acgacaacaa agactagtat tagtaccacc tcggctatcc acatagatca
      361 aagctggttt aaaagagttg tgcagatgat ccgtggcaga gacgagatct agtctgagtc
      421 gaccgtctct gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac
      481 ttgaaaaagt ggcaccgagt cggtgctttt tttgtccctt cgaagggcaa ttctgcagat
      541 atccatcaca ctggcggccg ctcgaggtcg agggtatcga taagcttgaa ttcgacccag
      601 ctttcttgta caaagttggc attataaaaa ataattgctc atcaatttgt tgcaacgaac
      661 aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg
      721 agtcgtatta catggtcata gcatccccta tagtgagtcg tattacatgg tcatagctgt
      781 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
      841 tatatcatca tgcctcctct ggaccagcca ggacagaaat gcctcgactt cgctgctgcc
      901 caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc agtggacata
      961 agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca actggtccag
     1021 aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga
     1081 ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt
     1141 gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa
     1201 aacaaagtta aacatcatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga
     1261 ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg
     1321 ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac
     1381 cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc
     1441 ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga
     1501 catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa
     1561 tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct
     1621 gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga
     1681 tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc
     1741 gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta
     1801 cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg
     1861 cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga
     1921 agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg
     1981 cgagatcacc aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa aatcccttaa
     2041 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2101 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2161 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2221 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     2281 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2341 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2401 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2461 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2521 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2581 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2641 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2701 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2761 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     2821 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     2881 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     2941 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctcgccagga
     3001 agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc
     3061 ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc
     3121 aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
     3181 aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc
     3241 tactctcgcg ttaacgcttg catggatgtt ttcccagtca cgacgttgta aaacgacggc
     3301 cagtcttaag ctcgggccca aataatgatt ttattttgac tgatagtgac ctgttcgttg
     3361 caacaaattg atgagcaatg cttttttata atgccaactt tgtatacaaa agttgcccca
     3421 tggcgttccc t
//
