LOCUS       sequence-391370-        3463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate entry vector; 7th gRNA under OsU6.2 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3463)
  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 3463)
  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: pCBLsgRNA7
FEATURES             Location/Qualifiers
     source          1..3463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..19)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(24..40)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(24..40)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      676..1264
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1165..1184
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1594..1621)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1713..1799)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(2023..2028,2029..2045,2046..2051)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             2099..3289
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(2451..2470)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(join(3463,1..19))
                     /label=T7
                     /note="T7 promoter, forward primer"
ORIGIN
        1 cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc
       61 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgcctc ctctagaggt
      121 ctcgcggtgg atcatgaacc aacggcctgg ctgtatttgg tggttgtgta gggagatggg
      181 gagaagaaaa gcccgattct cttcgctgtg atgggctgga tgcatgcggg ggagcgggag
      241 gcccaagtac gtgcacggtg agcggcccac agggcgagtg tgagcgcgag aggcgggagg
      301 aacagtttag taccacattg cccagctaac tcgaacgcga ccaacttata aacccgcgcg
      361 ctgtcgcttg tgttggagac gagatctagt ctgagtcgac cgtctctgtt tcagagctat
      421 gctggaaaca gcatagcaag ttgaaataag gctagtccgt tatcaacttg aaaaagtggc
      481 accgagtcgg tgcttttttt gtcccttcga agggcaattc tgcagatatc catcacactg
      541 gcggccgctc gaggtcgagg gtatcgataa gcttgaaagg agaccctcga gccacccatg
      601 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
      661 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
      721 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
      781 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
      841 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
      901 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
      961 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1021 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1081 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1141 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     1201 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     1261 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     1321 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     1381 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     1441 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     1501 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg
     1561 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
     1621 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
     1681 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
     1741 gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga
     1801 tgcctggcag ttccctactc tcgcgttctg cattaatgaa tcggccaacg cgcggggaga
     1861 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1921 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1981 tcaggggata acgcaggaaa gaacatgaat taattctcat gtttgacagc ttatcatcga
     2041 ttagctttaa tgcggtagtt tatcacagtt aaattgctaa cgcagtcagg caccgtgtat
     2101 gaaatctaac aatgcgctca tcgtcatcct cggcaccgtc accctggatg ctgtaggcat
     2161 aggcttggtt atgccggtac tgccgggcct cttgcgggat atcgtccatt ccgacagcat
     2221 cgccagtcac tatggcgtgc tgctagcgct atatgcgttg atgcaatttc tatgcgcacc
     2281 cgttctcgga gcactgtccg accgctttgg ccgccgccca gtcctgctcg cttcgctact
     2341 tggagccact atcgactacg cgatcatggc gaccacaccc gtcctgtgga ttctctacgc
     2401 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc
     2461 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg
     2521 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttacatgc
     2581 accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat
     2641 gcaggagtcg cataagggag agcgccgacc catgcccttg agagccttca acccagtcag
     2701 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
     2761 catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt
     2821 tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct
     2881 cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat
     2941 cgccggcatg gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg
     3001 gatggccttc cccattatga ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca
     3061 ggccatgctg tccaggcagg tagatgacga ccatcaggga cagcttcaag gatcgctcgc
     3121 ggctcttacc agcctaactt cgatcattgg accgctgatc gtcacggcga tttatgccgc
     3181 ctcggcgagc acatggaacg ggttggcatg gattgtaggc gccgccctat accttgtctg
     3241 cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa tggaagccgg
     3301 cggcacctcg ctaacggatt caccactcca agaattggag ccaatcaatt cttgcggaga
     3361 actgtgaatg cgcaaaccaa cccttgatcg gggaagaaca gtatgtcgag ctattttttg
     3421 acttactggg gatcaagcct gattgggaga aaataaaata tcc
//
