LOCUS       sequence-391240-        3580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate entry vector; 8th gRNA under TaU6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3580)
  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 3580)
  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: pCBLsgRNA8
FEATURES             Location/Qualifiers
     source          1..3580
                     /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      793..1381
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1282..1301
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1711..1738)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1830..1916)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(2140..2145,2146..2162,2163..2168)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             2216..3406
                     /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(2568..2587)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(join(3580,1..19))
                     /label=T7
                     /note="T7 promoter, forward primer"
ORIGIN
        1 cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc
       61 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgcctc ctctagaggt
      121 ctcggaaaga ccaagcccgt tattctgaca gttctggtgc tcaacacatt tatatttatc
      181 aaggagcaca ttgttactca ctgctaggag ggaatcgaac taggaatatt gatcagagga
      241 actacgagag agctgaagat aactgccctc tagctctcac tgatctgggt cgcatagtga
      301 gatgcagccc acgtgagttc agcaacggtc tagcgctggg cttttaggcc cgcatgatcg
      361 ggcttttgtc gggtggtcga cgtgttcacg attggggaga gcaacgcagc agttcctctt
      421 agtttagtcc cacctcgcct gtccagcaga gttctgaccg gtttataaac tcgcttgctg
      481 catcagactt gggagacgag atctagtctg agtcgaccgt ctctgtttca gagctatgct
      541 ggaaacagca tagcaagttg aaataaggct agtccgttat caacttgaaa aagtggcacc
      601 gagtcggtgc tttttttgtc ccttcgaagg gcaattctgc agatatccat cacactggcg
      661 gccgctcgag gtcgagggta tcgataagct ttcgaggaga ccctcgagcc acccatgacc
      721 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
      781 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
      841 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
      901 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
      961 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     1021 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     1081 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     1141 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     1201 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     1261 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     1321 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     1381 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     1441 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     1501 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     1561 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     1621 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac
     1681 cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc
     1741 ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc
     1801 ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac
     1861 aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc
     1921 ctggcagttc cctactctcg cgttctgcat taatgaatcg gccaacgcgc ggggagaggc
     1981 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2041 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2101 ggggataacg caggaaagaa catgaattaa ttctcatgtt tgacagctta tcatcgatta
     2161 gctttaatgc ggtagtttat cacagttaaa ttgctaacgc agtcaggcac cgtgtatgaa
     2221 atctaacaat gcgctcatcg tcatcctcgg caccgtcacc ctggatgctg taggcatagg
     2281 cttggttatg ccggtactgc cgggcctctt gcgggatatc gtccattccg acagcatcgc
     2341 cagtcactat ggcgtgctgc tagcgctata tgcgttgatg caatttctat gcgcacccgt
     2401 tctcggagca ctgtccgacc gctttggccg ccgcccagtc ctgctcgctt cgctacttgg
     2461 agccactatc gactacgcga tcatggcgac cacacccgtc ctgtggattc tctacgccgg
     2521 acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga
     2581 catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt
     2641 gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct tacatgcacc
     2701 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca
     2761 ggagtcgcat aagggagagc gccgacccat gcccttgaga gccttcaacc cagtcagctc
     2821 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     2881 gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg
     2941 ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc
     3001 tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc
     3061 cggcatggcg gccgacgcgc tgggctacgt cttgctggcg ttcgcgacgc gaggctggat
     3121 ggccttcccc attatgattc ttctcgcttc cggcggcatc gggatgcccg cgttgcaggc
     3181 catgctgtcc aggcaggtag atgacgacca tcagggacag cttcaaggat cgctcgcggc
     3241 tcttaccagc ctaacttcga tcattggacc gctgatcgtc acggcgattt atgccgcctc
     3301 ggcgagcaca tggaacgggt tggcatggat tgtaggcgcc gccctatacc ttgtctgcct
     3361 ccccgcgttg cgtcgcggtg catggagccg ggccacctcg acctgaatgg aagccggcgg
     3421 cacctcgcta acggattcac cactccaaga attggagcca atcaattctt gcggagaact
     3481 gtgaatgcgc aaaccaaccc ttgatcgggg aagaacagta tgtcgagcta ttttttgact
     3541 tactggggat caagcctgat tgggagaaaa taaaatatcc
//
