LOCUS       sequence-255111-        5063 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 5063)
  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 5063)
  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-Aac
FEATURES             Location/Qualifiers
     source          1..5063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1158..1253)
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1268..1285)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1268..1284)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1277..1299)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      1348..1434
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1526..1553
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1712..1729)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1883..2471)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1963..1982)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2564..3355)
                     /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"
     primer_bind     3786..3802
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3786..3802
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3807..3825
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        3842..3860
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3878..3977
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     ncRNA           complement(3997..4064)
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     ncRNA           complement(4189..4225)
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
ORIGIN
        1 agttagcaat cagaacgtgt ctgacgtaca ggtcgcatcc gtgtacgaac gctagcagca
       61 cggatctaac acaaacacgg atctaacaca aacatgaaca gaagtagaac taccgggccc
      121 taaccatgga ccggaacgcc gatctagaga aggtagagag gggggggggg gggaggacga
      181 gcggcgtacc ttgaagcgga ggtgccgacg ggtggatttg ggggagatct ggttgtgtgt
      241 gtgtgcgctc cgaacaacac gaggttgggg aaagagggtg tggagggggt gtctatttat
      301 tacggcgggc gaggaaggga aagcgaagga gcggtgggaa aggaatcccc cgtagctgcc
      361 ggtgccgtga gaggaggagg aggccgcctg ccgtgccggc tcacgtctgc cgctccgcca
      421 cgcaatttct ggatgccgac agcggagcaa gtccaacggt ggagcggaac tctcgagagg
      481 ggtccagagg cagcgacaga gatgccgtgc cgtctgcttc gcttggcccg acgcgacgct
      541 gctggttcgc tggttggtgt ccgttagact cgtcgacggc gtttaacagg ctggcattat
      601 ctactcgaaa caagaaaaat gtttccttag tttttttaat ttcttaaagg gtatttgttt
      661 aatttttagt cactttattt tattctattt tatatctaaa ttattaaata aaaaaactaa
      721 aatagagttt tagttttctt aatttagagg ctaaaataga ataaaataga tgtactaaaa
      781 aaattagtct ataaaaacca ttaaccctaa accctaaatg gatgtactaa taaaatggat
      841 gaagtattat ataggtgaag ctatttgcaa aaaaaaagga gaacacatgc acactaaaaa
      901 gataaaactg tagagtcctg ttgtcaaaat actcaattgt cctttagacc atgtctaact
      961 gttcatttat atgattctct aaaacactga tattattgta gtactataga ttatattatt
     1021 cgtagagtaa agtttaaata tatgtataaa gatagataaa ctgcacttca aacaagtgtg
     1081 acaaaaaaaa tatgtggtaa ttttttataa cttagacatg caatgctcat tatctctcta
     1141 gagggaacgc catggggcaa cttttgtata caaagttggc attataaaaa agcattgctc
     1201 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta ttgggcccga
     1261 gcttaagact ggccgtcgtt ttacaacgtc gtgactggga aaacatccat gcaagcgtta
     1321 acgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc ggaagactgg
     1381 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     1441 ggagcggatt tgaacgttgt gaagcaacgg cccggagggt ggcgggcagg acgcccgcca
     1501 taaactgcca ggcatcaaac taagcagaag gccatcctga cggatggcct ttttgcgttt
     1561 ctacaaactc ttcctggctc gcggtacgcg tattaattgc gttgcgctca ctgcccgctt
     1621 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     1681 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     1741 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1801 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1861 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1921 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1981 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2041 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2101 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2161 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2221 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2281 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2341 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2401 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2461 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgacg
     2521 cgtaactcac gttaagggat tttggtcatg ggtggctcga gggttatttg ccgactacct
     2581 tggtgatctc gcctttcacg tagtggacaa attcttccaa ctgatctgcg cgcgaggcca
     2641 agcgatcttc ttcttgtcca agataagcct gtctagcttc aagtatgacg ggctgatact
     2701 gggccggcag gcgctccatt gcccagtcgg cagcgacatc cttcggcgcg attttgccgg
     2761 ttactgcgct gtaccaaatg cgggacaacg taagcactac atttcgctca tcgccagccc
     2821 agtcgggcgg cgagttccat agcgttaagg tttcatttag cgcctcaaat agatcctgtt
     2881 caggaaccgg atcaaagagt tcctccgccg ctggacctac caaggcaacg ctatgttctc
     2941 ttgcttttgt cagcaagata gccagatcaa tgtcgatcgt ggctggctcg aagatacctg
     3001 caagaatgtc attgcgctgc cattctccaa attgcagttc gcgcttagct ggataacgcc
     3061 acggaatgat gtcgtcgtgc acaacaatgg tgacttctac agcgcggaga atctcgctct
     3121 ctccagggga agccgaagtt tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat
     3181 caagccttac ggtcaccgta accagcaaat caatatcact gtgtggcttc aggccgccat
     3241 ccactgcgga gccgtacaaa tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga
     3301 cgccaactac ctctgatagt tgagtcgata cttcggcgat caccgcttcc ctcataatgt
     3361 ttaactttgt tttagggcga ctgccctgct gcgtaacatc gttgctgctc cataacatca
     3421 aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata gactgtaccc
     3481 caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt
     3541 cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta cttgcattac agcttacgaa
     3601 ccgaacaggc ttatgtccac tgggttcgtg ccttcatccg tttccacggt gtgcgtcacc
     3661 cggcaacctt gggtagcagc gaagtcgagg catttctgtc ctggctggtc cagaggaggc
     3721 atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac acgggccaga
     3781 gctgccagga aacagctatg accatgtaat acgactcact ataggggatg ctatgaccat
     3841 gtaatacgac tcactatagg ggatatcagc tggatggcaa ataatgattt tattttgact
     3901 gatagtgacc tgttcgttgc aacaaattga tgagcaatta ttttttataa tgccaacttt
     3961 gtacaagaaa gctgggtcga attccggccg ggtaccgtcc cattcgccat gccgaagcat
     4021 gttgcccagc cggcgccagc gaggaggctg ggaccatgcc ggccagagac gccgcggtca
     4081 gactgagctc cgtctcgtgc cacttctcag atttgagaag ctcaacgggc tttgccacct
     4141 ggaaagtggc cattggcaca cccgttgaaa aattctgtcc tctagaccga cgagcttact
     4201 cgtttcgtcc tcacggactc atcagggtct aggatccctg cagaagtaac accaaacaac
     4261 agggtgagca tcgacaaaag aaacagtacc aagcaaataa atagcgtatg aaggcagggc
     4321 taaaaaaatc cacatatagc tgctgcatat gccatcatcc aagtatatca agatcaaaat
     4381 aattataaaa catacttgtt tattataata gataggtact caaggttaga gcatatgaat
     4441 agatgctgca tatgccatca tgtatatgca tcagtaaaac ccacatcaac atgtatacct
     4501 atcctagatc gatatttcca tccatcttaa actcgtaact atgaagatgt atgacacaca
     4561 catacagttc caaaattaat aaatacacca ggtagtttga aacagtattc tactccgatc
     4621 tagaacgaat gaacgaccgc ccaaccacac cacatcatca caaccaagcg aacaaaaagc
     4681 atctctgtat atgcatcagt aaaacccgca tcaacatgta tacctatcct agatcgatat
     4741 ttccatccat catcttcaat tcgtaactat gaatatgtat ggcacacaca tacagatcca
     4801 aaattaataa atccaccagg tagtttgaaa cagaattaat tctactccga tctagaacga
     4861 ccgcccaacc agaccacatc atcacaacca agacaaaaaa aagcatgaaa agatgacccg
     4921 acaaacaagt gcacggcata tattgaaata aaggaaaagg gcaaaccaaa ccctatgcaa
     4981 cgaaacaaaa aaaatcatga aatcgatccc gtctgcggaa cggctagagc catcccagga
     5041 ttccccaaag agaaacactg gca
//
