LOCUS       sequence-391201-        3440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for single gRNA under OsU3 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3440)
  AUTHORS   Lee K, Kang M, Ji Q, Grosic S, Wang K
  TITLE     New T-DNA binary vectors with NptII selection and RUBY reporter for 
            efficient maize transformation and targeted mutagenesis.
  JOURNAL   Plant Physiol. 2023 Apr 18:kiad231. doi: 10.1093/plphys/kiad231.
  PUBMED    37070560
REFERENCE   2  (bases 1 to 3440)
  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. 2023 Apr 18:kiad231. doi: 10.1093/plphys/kiad231."
COMMENT     Sequence Label: pCBLgRNA
FEATURES             Location/Qualifiers
     source          1..3440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(585..684)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(702..720)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(737..755)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(760..776)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(760..776)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1207..1998
                     /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      2091..2679
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2580..2599
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2833..2850
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3009..3036)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3128..3214)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3263..3285
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3277..3294
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3278..3294
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3309..3404
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 aagggatctt taaacatacg aacagatcac ttaaagttct tctgaagcaa cttaaagtta
       61 tcaggcatgc atggatcttg gaggaatcag atgtgcagtc agggaccata gcacaggaca
      121 ggcgtcttct actggtgcta ccagcaaatg ctggaagccg ggaacactgg gtacgttgga
      181 aaccacgtga tgtggagtaa gataaactgt aggagaaaag catttcgtag tgggccatga
      241 agcctttcag gacatgtatt gcagtatggg ccggcccatt acgcaattgg acgacaacaa
      301 agactagtat tagtaccacc tcggctatcc acatagatca aagctggttt aaaagagttg
      361 tgcagatgat ccgtggcaga gacgagatct agtctgagtc gaccgtctct gtttcagagc
      421 tatgctggaa acagcatagc aagttgaaat aaggctagtc cgttatcaac ttgaaaaagt
      481 ggcaccgagt cggtgctttt tttgtccctt cgaagggcaa ttctgcagat atccatcaca
      541 ctggcggccg ctcgaggtcg agggtatcga taagcttgaa ttcgacccag ctttcttgta
      601 caaagttggc attataaaaa ataattgctc atcaatttgt tgcaacgaac aggtcactat
      661 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
      721 catggtcata gcatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag
      781 ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca
      841 tgcctcctct ggaccagcca ggacagaaat gcctcgactt cgctgctacc caaggttgcc
      901 gggtgacgca caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg
      961 gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc
     1021 gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt
     1081 ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt
     1141 ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta
     1201 aacattatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc
     1261 gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg
     1321 gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt
     1381 gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga
     1441 gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg
     1501 tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt
     1561 gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca
     1621 agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct
     1681 gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac
     1741 tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta
     1801 accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc
     1861 cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc
     1921 ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc
     1981 aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa aatcccttaa cgtgagttac
     2041 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2101 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2161 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2221 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2281 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2341 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2401 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2461 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2521 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2581 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2641 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2701 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     2761 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     2821 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     2881 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     2941 aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg cgagccagga agagtttgta
     3001 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt
     3061 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
     3121 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     3181 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg
     3241 ttaacgcttg catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag
     3301 ctcgggccca ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga
     3361 tgagcaatgc ttttttataa tgccaacttt gtatacaaaa gttgccccat ggcgttccct
     3421 ctagataacg caggatccta
//
