LOCUS       sequence-123892-        3628 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector; single gRNA under AtU6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3628)
  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 3628)
  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: pYPQ141A
FEATURES             Location/Qualifiers
     source          1..3628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..324
                     /label=AtU6-1 promoter
                     /note="promoter for the Arabidopsis thaliana U6-1 snRNA 
                     gene (Waibel and Filipowicz, 1990)"
     misc_RNA        358..433
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(827..926)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(943..962)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(944..962)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(967..983)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(967..983)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1414..2205
                     /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      2298..2886
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2787..2806
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3040..3057
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3216..3243)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3335..3421)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3470..3492
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3484..3501
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3485..3501
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3517..3612
                     /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 agaaatctca aaattccggc agaacaattt tgaatctcga
       61 tccgtagaaa ccagacggtc attgttttag ttccaccacg attatatttg aaatttacgt
      121 gagtgtgagt gagacttgca taagaaaata aaatctttag ttgggaaaaa attcaataat
      181 ataaatgggc ttgagaagga agcgagggat aggccttttt ctaaaatagg cccatttaag
      241 ctattaacaa tcttcaaaag taccacagcg cttaggtaaa gaaagcagct gagtttatat
      301 atggttagac acgaagtagt gattggagac gagatctagt ctgagtcgac cgtctctgtt
      361 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
      421 accgagtcgg tgcttttttt ggcaaaaatt ttcagatttt ttcttcatct gtagatttct
      481 gggttttttt ttccgtttcg tgaatcataa gtgaagtttt ggatgcaaat ctgcgcgaaa
      541 aaagttggac ctgcaatgag cttatttaga tagctaagac aaagtgattg gtccgttgtt
      601 tcagttctga ttgtcagaga gtttgtttcg agtcggcgac accaatgcgt tttgttaacc
      661 agatttcggg taagaaatgt atcgagagtt tgtttccaga cggctacatc attttcttat
      721 gaagggtgaa attagataga ccaaagattg aaacacaaca tttctttcac aaaaatataa
      781 taaacttgat agcatttagg atcagcaact agtaagggcg aattcgaccc agctttcttg
      841 tacaaagttg gcattataaa aaataattgc tcatcaattt gttgcaacga acaggtcact
      901 atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat
      961 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
     1021 attgcacaag ataaaaatat atcatcatgc ctcctctgga ccagccagga cagaaatgcc
     1081 tcgacttcgc tgctgcccaa ggttgccggg tgacgcacac cgtggaaacg gatgaaggca
     1141 cgaacccagt ggacataagc ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc
     1201 tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt
     1261 ttcatggctt gttatgactg tttttttggg gtacagtcta tgcctcgggc atccaagcag
     1321 caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag
     1381 cagggcagtc gccctaaaac aaagttaaac atcatgaggg aagcggtgat cgccgaagta
     1441 tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg
     1501 gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat
     1561 ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac
     1621 cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc
     1681 attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt
     1741 ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt
     1801 gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg
     1861 gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc
     1921 ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg
     1981 ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc
     2041 gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag
     2101 gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt
     2161 gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataaccctc gagccaccca
     2221 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     2281 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2341 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2401 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2461 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2521 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2581 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2641 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2701 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2761 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2821 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2881 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2941 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3001 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3061 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     3121 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
     3181 cgtaccgctc gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     3241 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     3301 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     3361 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
     3421 gatgcctggc agttccctac tctcgcgtta acgcttgcat ggatgttttc ccagtcacga
     3481 cgttgtaaaa cgacggccag tcttaagctc gggcccaaat aatgatttta ttttgactga
     3541 tagtgacctg ttcgttgcaa caaattgatg agcaatgctt ttttataatg ccaactttgt
     3601 atacaaaagt tgccccatgg cgttccct
//
