LOCUS       sequence-375105-        3185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate entry vector to clone the 4th Cas12j2(Cas-Phi) crRNA 
            flanked by HH and HDV ribozymes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3185)
  AUTHORS   Liu S, Sretenovic S, Fan T, Cheng Y, Li G, Qi A, Tang X, Xu Y, Guo 
            W, Zhong Z, He Y, Liang Y, Han Q, Zheng X, Gu X, Qi Y, Zhang Y
  TITLE     Hypercompact CRISPR-Cas12j2 (CasPhi) enables genome editing, gene 
            activation, and epigenome editing in plants.
  JOURNAL   Plant Commun. 2022 Sep 20:100453. doi: 10.1016/j.xplc.2022.100453.
  PUBMED    36127876
REFERENCE   2  (bases 1 to 3185)
  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 
            Commun. 2022 Sep 20:100453. doi: 10.1016/j.xplc.2022.100453."
COMMENT     Sequence Label: pYPQ134-RZ-Cas12j2
FEATURES             Location/Qualifiers
     source          1..3185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(158..163,164..180,181..186)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             234..1424
                     /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(586..605)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(1598..1617)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1599..1617)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1622..1638)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1622..1638)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     ncRNA           1733..1769
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     ncRNA           1827..1894
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     rep_origin      1996..2584
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2485..2504
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2914..2941)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3033..3119)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
       61 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      121 ggataacgca ggaaagaaca tgaattaatt ctcatgtttg acagcttatc atcgattagc
      181 tttaatgcgg tagtttatca cagttaaatt gctaacgcag tcaggcaccg tgtatgaaat
      241 ctaacaatgc gctcatcgtc atcctcggca ccgtcaccct ggatgctgta ggcataggct
      301 tggttatgcc ggtactgccg ggcctcttgc gggatatcgt ccattccgac agcatcgcca
      361 gtcactatgg cgtgctgcta gcgctatatg cgttgatgca atttctatgc gcacccgttc
      421 tcggagcact gtccgaccgc tttggccgcc gcccagtcct gctcgcttcg ctacttggag
      481 ccactatcga ctacgcgatc atggcgacca cacccgtcct gtggattctc tacgccggac
      541 gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca
      601 tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg
      661 gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctcctta catgcaccat
      721 tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc ctaatgcagg
      781 agtcgcataa gggagagcgc cgacccatgc ccttgagagc cttcaaccca gtcagctcct
      841 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
      901 aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac cgctttcgct
      961 ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc
     1021 aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc attatcgccg
     1081 gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga ggctggatgg
     1141 ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca
     1201 tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggatcg ctcgcggctc
     1261 ttaccagcct aacttcgatc attggaccgc tgatcgtcac ggcgatttat gccgcctcgg
     1321 cgagcacatg gaacgggttg gcatggattg taggcgccgc cctatacctt gtctgcctcc
     1381 ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac ctgaatggaa gccggcggca
     1441 cctcgctaac ggattcacca ctccaagaat tggagccaat caattcttgc ggagaactgt
     1501 gaatgcgcaa accaaccctt gatcggggaa gaacagtatg tcgagctatt ttttgactta
     1561 ctggggatca agcctgattg ggagaaaata aaatatcccc tatagtgagt cgtattacat
     1621 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     1681 acaagataaa aatatatcat catgcctcct ctagaggtct cgccagtcgt tgctgatgag
     1741 tccgtgagga cgaaacgagt aagctcgtcc aacgattgcc cctcacgagg ggacgagacg
     1801 gagctcagtc tgaccgcggc gtctctggcc ggcatggtcc cagcctcctc gctggcgccg
     1861 gctgggcaac atgcttcggc atggcgaatg ggactgttgg agaccctcga gccacccatg
     1921 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     1981 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2041 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2101 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     2161 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2221 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2281 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2341 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2401 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2461 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2521 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2581 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2641 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2701 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2761 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     2821 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg
     2881 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
     2941 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
     3001 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
     3061 gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga
     3121 tgcctggcag ttccctactc tcgcgttctg cattaatgaa tcggccaacg cgcggggaga
     3181 ggcgg
//
