LOCUS       sequence-122130-        3298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector; single gRNA under OsU6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3298)
  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 3298)
  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: pYPQ141C
FEATURES             Location/Qualifiers
     source          1..3298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        333..408
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(497..596)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(613..632)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(614..632)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(637..653)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(637..653)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1084..1875
                     /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      1968..2556
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2457..2476
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2710..2727
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2886..2913)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3005..3091)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3140..3162
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3154..3171
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3155..3171
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3187..3282
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ctagataacg caggatccac tagtaacggc cgccagtgtg ctggaattgc ccttggatca
       61 tgaaccaacg gcctggctgt atttggtggt tgtgtaggga gatggggaga agaaaagccc
      121 gattctcttc gctgtgatgg gctggatgca tgcgggggag cgggaggccc aagtacgtgc
      181 acggtgagcg gcccacaggg cgagtgtgag cgcgagaggc gggaggaaca gtttagtacc
      241 acattgccca gctaactcga acgcgaccaa cttataaacc cgcgcgctgt cgcttgtgtg
      301 gagacgagat ctagtctgag tcgaccgtct ctgttttaga gctagaaata gcaagttaaa
      361 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttgtccc
      421 ttcgaagggc aattctgcag atatccatca cactggcggc cgctcgaggt cgagggtatc
      481 gataagcttg aattcgaccc agctttcttg tacaaagttg gcattataaa aaataattgc
      541 tcatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat
      601 ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc
      661 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatgc
      721 ctcctctgga ccagccagga cagaaatgcc tcgacttcgc tgctgcccaa ggttgccggg
      781 tgacgcacac cgtggaaacg gatgaaggca cgaacccagt ggacataagc ctgttcggtt
      841 cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa
      901 cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg tttttttggg
      961 gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg
     1021 atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac aaagttaaac
     1081 atcatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt agttggcgtc
     1141 atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc cgcagtggat
     1201 ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt aaggcttgat
     1261 gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc ccctggagag
     1321 agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat cattccgtgg
     1381 cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga cattcttgca
     1441 ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac aaaagcaaga
     1501 gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc ggttcctgaa
     1561 caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg
     1621 gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag cgcagtaacc
     1681 ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct gccggcccag
     1741 tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga agatcgcttg
     1801 gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga gatcaccaag
     1861 gtagtcggca aataaccctc gagccaccca tgaccaaaat cccttaacgt gagttacgcg
     1921 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     1981 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2041 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2101 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2161 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2221 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2281 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2341 agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2401 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2461 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     2521 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     2581 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     2641 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     2701 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     2761 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     2821 gcgggcagtg agcgcaacgc aattaatacg cgtaccgctc gccaggaaga gtttgtagaa
     2881 acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg
     2941 gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg
     3001 gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc
     3061 agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcgtta
     3121 acgcttgcat ggatgttttc ccagtcacga cgttgtaaaa cgacggccag tcttaagctc
     3181 gggcccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgatg
     3241 agcaatgctt ttttataatg ccaactttgt atacaaaagt tgccccatgg cgttccct
//
