LOCUS       sequence-391286-        3547 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate entry vector; 6th gRNA under OsU6.1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3547)
  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 3547)
  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: pCBLsgRNA6
FEATURES             Location/Qualifiers
     source          1..3547
                     /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      760..1348
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1249..1268
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1678..1705)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1797..1883)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(2107..2112,2113..2129,2130..2135)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             2183..3373
                     /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(2535..2554)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(join(3547,1..19))
                     /label=T7
                     /note="T7 promoter, forward primer"
ORIGIN
        1 cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc
       61 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgcctc ctctagaggt
      121 ctcgtgcaaa gaacgaacta agccggacaa aaaaaaaaag gagcacatat acaaaccggt
      181 tttattcatg aatggtcacg atggatgatg gggctcagac ttgagctacg aggccgcagg
      241 cgagagaagc ctagtgtgct ctctgcttgt ttgggccgta acggaggata cggccgacga
      301 gcgtgtacta ccgcgcggga tgccgctggg cgctgcgggg gccgttggat ggggatcggt
      361 gggtcgcggg agcgttgagg ggagacaggt ttagtaccac ctcgcctacc gaacaatgaa
      421 gaacccacct tataaccccg cgcgctgccg cttgtgttgg agacgagatc tagtctgagt
      481 cgaccgtctc tgtttcagag ctatgctgga aacagcatag caagttgaaa taaggctagt
      541 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttgtccct tcgaagggca
      601 attctgcaga tatccatcac actggcggcc gctcgaggtc gagggtatcg ataagcttcg
      661 gtggagaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg cgtcgttcca
      721 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
      781 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
      841 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
      901 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
      961 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1021 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1081 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1141 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1201 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1261 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1321 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1381 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     1441 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     1501 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     1561 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     1621 tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa
     1681 aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt
     1741 cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt
     1801 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc
     1861 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt tctgcattaa
     1921 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     1981 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2041 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gaattaattc
     2101 tcatgtttga cagcttatca tcgattagct ttaatgcggt agtttatcac agttaaattg
     2161 ctaacgcagt caggcaccgt gtatgaaatc taacaatgcg ctcatcgtca tcctcggcac
     2221 cgtcaccctg gatgctgtag gcataggctt ggttatgccg gtactgccgg gcctcttgcg
     2281 ggatatcgtc cattccgaca gcatcgccag tcactatggc gtgctgctag cgctatatgc
     2341 gttgatgcaa tttctatgcg cacccgttct cggagcactg tccgaccgct ttggccgccg
     2401 cccagtcctg ctcgcttcgc tacttggagc cactatcgac tacgcgatca tggcgaccac
     2461 acccgtcctg tggattctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg
     2521 tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt
     2581 cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact
     2641 gttgggcgcc atctccttac atgcaccatt ccttgcggcg gcggtgctca acggcctcaa
     2701 cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgcc gacccatgcc
     2761 cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc
     2821 cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctg
     2881 ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc
     2941 ggtattcgga atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg
     3001 tttcggcgag aagcaggcca ttatcgccgg catggcggcc gacgcgctgg gctacgtctt
     3061 gctggcgttc gcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg
     3121 cggcatcggg atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca
     3181 gggacagctt caaggatcgc tcgcggctct taccagccta acttcgatca ttggaccgct
     3241 gatcgtcacg gcgatttatg ccgcctcggc gagcacatgg aacgggttgg catggattgt
     3301 aggcgccgcc ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc
     3361 cacctcgacc tgaatggaag ccggcggcac ctcgctaacg gattcaccac tccaagaatt
     3421 ggagccaatc aattcttgcg gagaactgtg aatgcgcaaa ccaacccttg atcggggaag
     3481 aacagtatgt cgagctattt tttgacttac tggggatcaa gcctgattgg gagaaaataa
     3541 aatatcc
//
