LOCUS       sequence-245053-        7390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector with pcoCas9-x3.7 (Plant codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7390)
  AUTHORS   Zhong Z, Sretenovic S, Ren Q, Yang L, Bao Y, Qi C, Yuan M, He Y, Liu
            S, Liu X, Wang J, Huang L, Wang Y, Baby D, Wang D, Zhang T, Qi Y, 
            Zhang Y
  TITLE     Improving Plant Genome Editing with High-Fidelity xCas9 and 
            Non-canonical PAM-Targeting Cas9-NG.
  JOURNAL   Mol Plant. 2019 Mar 27. pii: S1674-2052(19)30124-8. doi: 
            10.1016/j.molp.2019.03.011.
  PUBMED    30928637
REFERENCE   2  (bases 1 to 7390)
  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: "Mol Plant. 
            2019 Mar 27. pii: S1674-2052(19)30124-8. doi: 
            10.1016/j.molp.2019.03.011."
COMMENT     Sequence Label: pYPQ150-x3.7
FEATURES             Location/Qualifiers
     source          1..7390
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(62..148)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     197..219
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     211..228
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     212..228
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    244..343
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             365..412
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             419..439
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4565..4612
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      4623..4875
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    4920..5043
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5060..5079)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5061..5079)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5084..5100)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5084..5100)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5531..6322
                     /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      6415..7003
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6904..6923
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7157..7174
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(7333..7360)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg
       61 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
      121 cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg
      181 cttgcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct taagctcggg
      241 ccccaaataa tgattttatt ttgactgata gtgacctgtt cgttgcaaca aattgatgag
      301 caatgctttt ttataatgcc aactttgtac aaaaaagcag gctccgaatt cgcccttcac
      361 catggattac aaggatgatg atgataagga ttacaaggat gatgatgata agatggctcc
      421 aaagaagaag agaaaggttg gaatccacgg agttccagct gctgataaga agtactctat
      481 cggacttgac atcggaacca actctgttgg atgggctgtt atcaccgatg agtacaaggt
      541 tccatctaag aagttcaagg ttcttggaaa caccgataga cactctatca agaagaacct
      601 tatcggtgct cttcttttcg attctggaga gaccgctgag gctaccagat tgaagagaac
      661 cgctagaaga agatacacca gaagaaagaa cagaatctgc taccttcagg aaatcttctc
      721 taacgagatg gctaaggttg atgattcttt cttccacaga cttgaggagt ctttccttgt
      781 tgaggaggat aagaagcacg agagacaccc aatcttcgga aacatcgttg atgaggttgc
      841 ttaccacgag aagtacccaa ccatctacca ccttagaaag aagttggttg attctaccga
      901 taaggctgat cttagactta tctaccttgc tcttgctcac atgatcaagt tcagaggaca
      961 cttccttatc gagggagacc ttaacccaga taactctgat gttgataagt tgttcatcca
     1021 gcttgttcag acctacaacc agcttttcga ggagaaccca atcaacgctt ctggagttga
     1081 tgctaaggct atcctttctg ctagactttc taagtctcgt agacttgaga accttatcgc
     1141 tcagcttcca ggagagaaga agaacggact tttcggaaac cttatcgctc tttctcttgg
     1201 acttacccca aacttcaagt ctaacttcga tcttgctgag gataccaagt tgcagctttc
     1261 taaggatacc tacgatgatg atcttgataa ccttcttgct cagatcggag atcagtacgc
     1321 tgatcttttc cttgctgcta agaacctttc tgatgctatc cttctttctg acatccttag
     1381 agttaacacc gagatcacca aggctccact ttctgcttct atgatcaagc tctacgatga
     1441 gcaccaccag gatcttaccc ttttgaaggc tcttgttaga cagcagcttc cagagaagta
     1501 caaggaaatc ttcttcgatc agtctaagaa cggatacgct ggatacatcg atggaggagc
     1561 ttctcaggag gagttctaca agttcatcaa gccaatcctt gagaagatgg atggaaccga
     1621 ggagcttctt gttaagttga acagagagga tcttcttaga aagcagagaa ccttcgataa
     1681 cggaatcatc ccacaccaga tccaccttgg agagcttcac gctatccttc gtagacagga
     1741 ggatttctac ccattcttga aggataacag agagaagatc gagaagatcc ttaccttcag
     1801 aatcccatac tacgttggac cacttgctag aggaaactct agtttcgctt ggatgaccag
     1861 aaagtctgag gagaccatca ccccttggaa cttcgagaag gttgttgata agggagcttc
     1921 tgctcagtct ttcatcgaga gaatgaccaa cttcgataag aaccttccaa acgagaaggt
     1981 tcttccaaag cactctcttc tttacgagta cttcaccgtt tacaacgagc ttaccaaggt
     2041 taagtacgtt accgagggaa tgagaaagcc agctttcctt tctggagacc agaagaaggc
     2101 tatcgttgat cttcttttca agaccaacag aaaggttacc gttaagcagt tgaaggagga
     2161 ttacttcaag aagatcgagt gcttcgattc tgttgaaatc tctggagttg aggatagatt
     2221 caacgcttct cttggaacct accacgatct tttgaagatc atcaaagata aggacttcct
     2281 tgataacgaa gagaacgagg atatcctcga ggacatcgtt cttaccttga cccttttcga
     2341 ggatagagag atgatcgagg agagactcaa gacctacgct caccttttcg atgataaggt
     2401 tatgaagcag ttgaagagaa gaagatacac cggatggggt agactttctc gtaagttgat
     2461 caacggaatc agagataagc agtctggaaa gaccatcctt gatttcttga agtctgatgg
     2521 attcgctaac agaaacttca tccagcttat ccacgatgat tctcttacct tcaaggagga
     2581 catccagaag gctcaggttt ctggacaggg agattctctt cacgagcaca tcgctaacct
     2641 tgctggatct ccagctatca agaagggaat ccttcagacc gttaaggttg ttgatgagct
     2701 tgttaaggtt atgggtagac acaagccaga gaacatcgtt atcgagatgg ctagagagaa
     2761 ccagaccacc cagaagggac agaagaactc tcgtgagaga atgaagagaa tcgaggaggg
     2821 aatcaaggag cttggatctc aaatcttgaa ggagcaccca gttgagaaca cccagcttca
     2881 gaacgagaag ttgtaccttt actaccttca gaacggaaga gatatgtacg ttgatcagga
     2941 gcttgacatc aacagacttt ctgattacga tgttgatcac atcgttccac agtctttctt
     3001 gaaggatgat tctatcgata acaaggttct tacccgttct gataagaaca gaggaaagtc
     3061 tgataacgtt ccatctgagg aggttgttaa gaagatgaag aactactgga gacagcttct
     3121 taacgctaag ttgatcaccc agagaaagtt cgataacctt accaaggctg agagaggagg
     3181 actttctgag cttgataagg ctggattcat caagagacag cttgttgaga ccagacagat
     3241 caccaagcac gttgctcaga tccttgattc tcgtatgaac accaagtacg atgagaacga
     3301 taagttgatc agagaggtta aggttatcac cttgaagtct aagttggttt ctgatttcag
     3361 aaaggatttc cagttctaca aggttagaga gatcaacaac taccaccacg ctcacgatgc
     3421 ttaccttaac gctgttgttg gaaccgctct tatcaagaag tacccaaagt tggagtctga
     3481 gttcgtttac ggagattaca aggtttacga tgttagaaag atgatcgcta agtctgagca
     3541 ggagatcgga aaggctaccg ctaagtactt cttctactct aacatcatga acttcttcaa
     3601 gaccgagatc acccttgcta acggagagat cagaaagaga ccacttatcg agaccaacgg
     3661 agagaccgga gagatcgttt gggataaggg aagagatttc gctaccgtta gaaaggttct
     3721 ttctatgcca caggttaaca tcgttaagaa aaccgaggtt cagaccggag gattctctaa
     3781 ggagtctatc cttccaaaga gaaactctga taagttgatc gctagaaaga aggattggga
     3841 cccaaagaag tacggaggat tcgattctcc aaccgttgct tactctgttc ttgttgttgc
     3901 taaggttgag aagggaaagt ctaagaagtt gaagtctgtt aaggagcttc ttggaatcac
     3961 catcatggag cgttcttctt tcgagaagaa cccaatcgat ttccttgagg ctaagggata
     4021 caaggaggtt aagaaggatc ttatcatcaa gttgccaaag tactctcttt tcgagcttga
     4081 gaacggaaga aagagaatgc ttgcttctgc tggagtgctt cagaagggaa acgagcttgc
     4141 tcttccatct aagtacgtta acttccttta ccttgcttct cactacgaga agttgaaggg
     4201 atctccagag gataacgagc agaagcagct tttcgttgag cagcacaagc actaccttga
     4261 tgagatcatc gagcaaatct ctgagttctc taagagagtt atccttgctg atgctaacct
     4321 tgataaggtt ctttctgctt acaacaagca cagagataag ccaatcagag agcaggctga
     4381 gaacatcatc caccttttca cccttaccaa ccttggtgct ccagctgctt tcaagtactt
     4441 cgataccacc atcgatagaa aaagatacac ctctaccaag gaggttcttg atgctaccct
     4501 tatccaccag tctatcaccg gactttacga gaccagaatc gatctttctc agcttggagg
     4561 agataagaga ccagctgcta ccaagaaggc tggacaggct aagaagaaga agtgagacgt
     4621 ccgatcgttc aaacatttgg caataaagtt tcttaagatt gaatcctgtt gccggtcttg
     4681 cgatgattat catataattt ctgttgaatt acgttaagca tgtaataatt aacatgtaat
     4741 gcatgacgtt atttatgaga tgggttttta tgattagagt cccgcaatta tacatttaat
     4801 acgcgataga aaacaaaata tagcgcgcaa actaggataa attatcgcgc gcggtgtcat
     4861 ctatgttact agatcgggaa ttgatccccc ctcgacagct tccggaaagg gcgaattcgc
     4921 aactttgtat acaaaagttg aacgagaaac gtaaaatgat ataaatatca atatattaaa
     4981 ttagattttg cataaaaaac agactacata atactgtaaa acacaacata tccagtcact
     5041 atgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     5101 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     5161 atcatgcctc ctctggacca gccaggacag aaatgcctcg acttcgctgc tacccaaggt
     5221 tgccgggtga cgcacaccgt ggaaacggat gaaggcacga acccagtgga cataagcctg
     5281 ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca cgcaactggt ccagaacctt
     5341 gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt
     5401 ttttggggta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg
     5461 atgtttgatg ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa
     5521 gttaaacatt atgagggaag cggtgatcgc cgaagtatcg actcaactat cagaggtagt
     5581 tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt acggctccgc
     5641 agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg tgaccgtaag
     5701 gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt cggcttcccc
     5761 tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg acgacatcat
     5821 tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc gcaatgacat
     5881 tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct tgctgacaaa
     5941 agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct ttgatccggt
     6001 tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga actcgccgcc
     6061 cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc
     6121 agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc
     6181 ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac aagaagaaga
     6241 tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga aaggcgagat
     6301 caccaaggta gtcggcaaat aaccctcgag ccacccatga ccaaaatccc ttaacgtgag
     6361 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6421 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6481 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6541 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6601 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6661 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6721 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6781 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6841 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6901 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6961 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7021 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7081 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     7141 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     7201 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     7261 ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgcgagcc aggaagagtt
     7321 tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca
     7381 gtttatggcg
//
