LOCUS       sequence-255058-        5072 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  (Empty backbone) Gateway entry clone with sgRNA under ZmUbi 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5072)
  AUTHORS   Ming M, Ren Q, Pan C, He Y, Zhang Y, Liu S, Zhong Z, Wang J, Malzahn
            AA, Wu J, Zheng X, Zhang Y, Qi Y
  TITLE     CRISPR-Cas12b enables efficient plant genome engineering.
  JOURNAL   Nat Plants. 2020 Mar;6(3):202-208. doi: 10.1038/s41477-020-0614-6. 
            Epub 2020 Mar 9.
  PUBMED    32170285
REFERENCE   2  (bases 1 to 5072)
  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"; doi: 
            "10.1038/s41477-020-0614-6"; journalName: "Nat Plants"; date: 
            "2020-03"; volume: "6"; issue: "3"; pages: "202-208"
COMMENT     Sequence Label: pYPQ141-ZmUbi-RZ-Bth
FEATURES             Location/Qualifiers
     source          1..5072
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     ncRNA           1013..1049
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     ncRNA           1186..1253
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     protein_bind    complement(1273..1372)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1390..1408)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1425..1443)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1448..1464)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1448..1464)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1895..2686
                     /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      2779..3367
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3268..3287
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3521..3538
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3697..3724)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3816..3902)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3951..3973
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3965..3982
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3966..3982
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3997..4092
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 tcctcccccc ccccctctct accttctcta gatcggcgtt ccggtccatg gttagggccc
       61 ggtagttcta cttctgttca tgtttgtgtt agatccgtgt ttgtgttaga tccgtgctgc
      121 tagcgttcgt acacggatgc gacctgtacg tcagacacgt tctgattgct aacttgccag
      181 tgtttctctt tggggaatcc tgggatggct ctagccgttc cgcagacggg atcgatttca
      241 tgattttttt tgtttcgttg catagggttt ggtttgccct tttcctttat ttcaatatat
      301 gccgtgcact tgtttgtcgg gtcatctttt catgcttttt tttgtcttgg ttgtgatgat
      361 gtggtctggt tgggcggtcg ttctagatcg gagtagaatt aattctgttt caaactacct
      421 ggtggattta ttaattttgg atctgtatgt gtgtgccata catattcata gttacgaatt
      481 gaagatgatg gatggaaata tcgatctagg ataggtatac atgttgatgc gggttttact
      541 gatgcatata cagagatgct ttttgttcgc ttggttgtga tgatgtggtg tggttgggcg
      601 gtcgttcatt cgttctagat cggagtagaa tactgtttca aactacctgg tgtatttatt
      661 aattttggaa ctgtatgtgt gtgtcataca tcttcatagt tacgagttta agatggatgg
      721 aaatatcgat ctaggatagg tatacatgtt gatgtgggtt ttactgatgc atatacatga
      781 tggcatatgc agcatctatt catatgctct aaccttgagt acctatctat tataataaac
      841 aagtatgttt tataattatt ttgatcttga tatacttgga tgatggcata tgcagcagct
      901 atatgtggat ttttttagcc ctgccttcat acgctattta tttgcttggt actgtttctt
      961 ttgtcgatgc tcaccctgtt gtttggtgtt acttctgcag ggatccctcg ccctgatgag
     1021 tccgtgagga cgaaacgagt aagctcgtcg gcgaggttct gtcttttggt caggacaacc
     1081 gtctagctat aagtgctgca ggggtgtgag aaactcctat tgctggacga tgtctctttc
     1141 gaggcattag cacgagacgg agctcagtct gaccgcggcg tctctggccg gcatggtccc
     1201 agcctcctcg ctggcgccgg ctgggcaaca tgcttcggca tggcgaatgg gacggtaccc
     1261 ggccggaatt cgacccagct ttcttgtaca aagttggcat tataaaaaat aattgctcat
     1321 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag
     1381 ctgatatccc ctatagtgag tcgtattaca tggtcatagc atcccctata gtgagtcgta
     1441 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     1501 cattgcacaa gataaaaata tatcatcatg cctcctctgg accagccagg acagaaatgc
     1561 ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc
     1621 acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg
     1681 ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt
     1741 tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca
     1801 gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca
     1861 gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga tcgccgaagt
     1921 atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct
     1981 ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga
     2041 tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga
     2101 ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac
     2161 cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt
     2221 tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat
     2281 tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc
     2341 ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac
     2401 cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac
     2461 gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc
     2521 cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca
     2581 ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt
     2641 tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct cgagccaccc
     2701 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     2761 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2821 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2881 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2941 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3001 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3061 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3121 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3181 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3241 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3301 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3361 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3421 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3481 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3541 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     3601 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac
     3661 gcgtaccgcg agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc
     3721 ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc
     3781 cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc
     3841 accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt
     3901 tgatgcctgg cagttcccta ctctcgcgtt aacgcttgca tggatgtttt cccagtcacg
     3961 acgttgtaaa acgacggcca gtcttaagct cgggcccaat aatgatttta ttttgactga
     4021 tagtgacctg ttcgttgcaa caaattgatg agcaatgctt ttttataatg ccaactttgt
     4081 atacaaaagt tgccccatgg cgttccctct agagagataa tgagcattgc atgtctaagt
     4141 tataaaaaat taccacatat tttttttgtc acacttgttt gaagtgcagt ttatctatct
     4201 ttatacatat atttaaactt tactctacga ataatataat ctatagtact acaataatat
     4261 cagtgtttta gagaatcata taaatgaaca gttagacatg gtctaaagga caattgagta
     4321 ttttgacaac aggactctac agttttatct ttttagtgtg catgtgttct cctttttttt
     4381 tgcaaatagc ttcacctata taatacttca tccattttat tagtacatcc atttagggtt
     4441 tagggttaat ggtttttata gactaatttt tttagtacat ctattttatt ctattttagc
     4501 ctctaaatta agaaaactaa aactctattt tagttttttt atttaataat ttagatataa
     4561 aatagaataa aataaagtga ctaaaaatta aacaaatacc ctttaagaaa ttaaaaaaac
     4621 taaggaaaca tttttcttgt ttcgagtaga taatgccagc ctgttaaacg ccgtcgacga
     4681 gtctaacgga caccaaccag cgaaccagca gcgtcgcgtc gggccaagcg aagcagacgg
     4741 cacggcatct ctgtcgctgc ctctggaccc ctctcgagag ttccgctcca ccgttggact
     4801 tgctccgctg tcggcatcca gaaattgcgt ggcggagcgg cagacgtgag ccggcacggc
     4861 aggcggcctc ctcctcctct cacggcaccg gcagctacgg gggattcctt tcccaccgct
     4921 ccttcgcttt cccttcctcg cccgccgtaa taaatagaca ccccctccac accctctttc
     4981 cccaacctcg tgttgttcgg agcgcacaca cacacaacca gatctccccc aaatccaccc
     5041 gtcggcacct ccgcttcaag gtacgccgct cg
//
