LOCUS       sequence-391371-        3411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate entry vector; 5th gRNA under ZmU6-C3 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3411)
  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 3411)
  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: pCBLsgRNA5
FEATURES             Location/Qualifiers
     source          1..3411
                     /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      624..1212
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1113..1132
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1542..1569)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1661..1747)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(1971..1976,1977..1993,1994..1999)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             2047..3237
                     /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(2399..2418)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(join(3411,1..19))
                     /label=T7
                     /note="T7 promoter, forward primer"
ORIGIN
        1 cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc
       61 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgcctc ctctagaggt
      121 ctcgtgttca aaaacatcct cacaggaaag acacgaagaa acatggtcaa tggcccatta
      181 tataaagcac cgccacaaag cccaaatacc agttcgtcgg tggagcaagt aacgcgctag
      241 gcaacaggca aacagtttgt cccacctcgt ccagtcacaa aggcaaagcg tgacttataa
      301 gccagagcgg aagaaccata ccggagacga gatctagtct gagtcgaccg tctctgtttc
      361 agagctatgc tggaaacagc atagcaagtt gaaataaggc tagtccgtta tcaacttgaa
      421 aaagtggcac cgagtcggtg ctttttttgt cccttcgaag ggcaattctg cagatatcca
      481 tcacactggc ggccgctcga ggtcgagggt atcgataagc tttgcaggag accctcgagc
      541 cacccatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
      601 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
      661 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
      721 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
      781 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
      841 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
      901 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
      961 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     1021 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     1081 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     1141 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     1201 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     1261 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     1321 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     1381 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     1441 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     1501 aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
     1561 tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
     1621 cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     1681 cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt
     1741 ttatttgatg cctggcagtt ccctactctc gcgttctgca ttaatgaatc ggccaacgcg
     1801 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     1861 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1921 ccacagaatc aggggataac gcaggaaaga acatgaatta attctcatgt ttgacagctt
     1981 atcatcgatt agctttaatg cggtagttta tcacagttaa attgctaacg cagtcaggca
     2041 ccgtgtatga aatctaacaa tgcgctcatc gtcatcctcg gcaccgtcac cctggatgct
     2101 gtaggcatag gcttggttat gccggtactg ccgggcctct tgcgggatat cgtccattcc
     2161 gacagcatcg ccagtcacta tggcgtgctg ctagcgctat atgcgttgat gcaatttcta
     2221 tgcgcacccg ttctcggagc actgtccgac cgctttggcc gccgcccagt cctgctcgct
     2281 tcgctacttg gagccactat cgactacgcg atcatggcga ccacacccgt cctgtggatt
     2341 ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc
     2401 tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct
     2461 tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc
     2521 ttacatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc
     2581 ttcctaatgc aggagtcgca taagggagag cgccgaccca tgcccttgag agccttcaac
     2641 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc
     2701 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag
     2761 gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg
     2821 cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag
     2881 gccattatcg ccggcatggc ggccgacgcg ctgggctacg tcttgctggc gttcgcgacg
     2941 cgaggctgga tggccttccc cattatgatt cttctcgctt ccggcggcat cgggatgccc
     3001 gcgttgcagg ccatgctgtc caggcaggta gatgacgacc atcagggaca gcttcaagga
     3061 tcgctcgcgg ctcttaccag cctaacttcg atcattggac cgctgatcgt cacggcgatt
     3121 tatgccgcct cggcgagcac atggaacggg ttggcatgga ttgtaggcgc cgccctatac
     3181 cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc gacctgaatg
     3241 gaagccggcg gcacctcgct aacggattca ccactccaag aattggagcc aatcaattct
     3301 tgcggagaac tgtgaatgcg caaaccaacc cttgatcggg gaagaacagt atgtcgagct
     3361 attttttgac ttactgggga tcaagcctga ttgggagaaa ataaaatatc c
//
