LOCUS       sequence-123890-        3459 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector; single gRNA under AtU3 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3459)
  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 3459)
  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: pYPQ141B
FEATURES             Location/Qualifiers
     source          1..3459
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        380..455
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(658..757)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(774..793)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(775..793)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(798..814)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(798..814)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1245..2036
                     /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      2129..2717
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2618..2637
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2871..2888
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3047..3074)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3166..3252)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3301..3323
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3315..3332
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3316..3332
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3348..3443
                     /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 tttactttaa atttttctta tggctcagcc tgtgatggat
       61 aactgaatca aacaaatggc gtctgggttt aagaacatct gttttggcta tgttggacga
      121 aacaagtgaa cttttaggat caacttccgt ttatatacgg agcttatatc gagcaataag
      181 ataagtgggc tttttatgta atttaatggg ctatcgtcca tatattcact aatacccatg
      241 cccagtaccc atgtatgcgt ttcatataag ctcctaattt ctcccacatc gctcaaatct
      301 aaacaaatct tgttgtatat ataacactga gggagcacca ttggtcagag acgagatcta
      361 gtctgagtcg accgtctctg ttttagagct agaaatagca agttaaaata aggctagtcc
      421 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tttccctttc cttttttctt
      481 ttttttgcca taaacttaaa tttgtatatc gatcattgta gatattgaaa acctagaaca
      541 aaccaacatc catgtgaatg tctttcatga ctgatttaga gataattctt gaattttgga
      601 actagaatct ataatgagcc taaattaaaa cattgtgaac tagtaagggc gaattcgacc
      661 cagctttctt gtacaaagtt ggcattataa aaaataattg ctcatcaatt tgttgcaacg
      721 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata
      781 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
      841 tctgatgtta cattgcacaa gataaaaata tatcatcatg cctcctctgg accagccagg
      901 acagaaatgc ctcgacttcg ctgctgccca aggttgccgg gtgacgcaca ccgtggaaac
      961 ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt
     1021 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg
     1081 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg
     1141 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
     1201 atgttacgca gcagggcagt cgccctaaaa caaagttaaa catcatgagg gaagcggtga
     1261 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
     1321 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
     1381 gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
     1441 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
     1501 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
     1561 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
     1621 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
     1681 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
     1741 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
     1801 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
     1861 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
     1921 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
     1981 tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct
     2041 cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     2101 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2161 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2221 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     2281 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2341 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2401 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2461 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2521 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2581 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2641 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2701 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     2761 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     2821 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     2881 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     2941 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     3001 caattaatac gcgtaccgct cgccaggaag agtttgtaga aacgcaaaaa ggccatccgt
     3061 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
     3121 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
     3181 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
     3241 tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgcttgca tggatgtttt
     3301 cccagtcacg acgttgtaaa acgacggcca gtcttaagct cgggcccaaa taatgatttt
     3361 attttgactg atagtgacct gttcgttgca acaaattgat gagcaatgct tttttataat
     3421 gccaactttg tatacaaaag ttgccccatg gcgttccct
//
