LOCUS       sequence-375102-        3185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate entry vector to clone the 1st 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: pYPQ131-RZ-Cas12j2
FEATURES             Location/Qualifiers
     source          1..3185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(202..207,208..224,225..230)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             278..1468
                     /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(630..649)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(1642..1661)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1643..1661)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1666..1682)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1666..1682)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     ncRNA           1777..1813
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     ncRNA           1871..1938
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     rep_origin      2040..2628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2529..2548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2958..2985)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3077..3163)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cgcgttctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
       61 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
      121 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      181 aacatgaatt aattctcatg tttgacagct tatcatcgat tagctttaat gcggtagttt
      241 atcacagtta aattgctaac gcagtcaggc accgtgtatg aaatctaaca atgcgctcat
      301 cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata ggcttggtta tgccggtact
      361 gccgggcctc ttgcgggata tcgtccattc cgacagcatc gccagtcact atggcgtgct
      421 gctagcgcta tatgcgttga tgcaatttct atgcgcaccc gttctcggag cactgtccga
      481 ccgctttggc cgccgcccag tcctgctcgc ttcgctactt ggagccacta tcgactacgc
      541 gatcatggcg accacacccg tcctgtggat tctctacgcc ggacgcatcg tggccggcat
      601 caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga
      661 tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc
      721 cgtggccggg ggactgttgg gcgccatctc cttacatgca ccattccttg cggcggcggt
      781 gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga
      841 gcgccgaccc atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
      901 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
      961 gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat
     1021 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg
     1081 tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc
     1141 gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat
     1201 tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt
     1261 agatgacgac catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc
     1321 gatcattgga ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg
     1381 gttggcatgg attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg
     1441 tgcatggagc cgggccacct cgacctgaat ggaagccggc ggcacctcgc taacggattc
     1501 accactccaa gaattggagc caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac
     1561 ccttgatcgg ggaagaacag tatgtcgagc tattttttga cttactgggg atcaagcctg
     1621 attgggagaa aataaaatat cccctatagt gagtcgtatt acatggtcat agctgtttcc
     1681 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     1741 tcatcatgcc tcctctagag gtctcgctat tcgttgctga tgagtccgtg aggacgaaac
     1801 gagtaagctc gtccaacgat tgcccctcac gaggggacga gacggagctc agtctgaccg
     1861 cggcgtctct ggccggcatg gtcccagcct cctcgctggc gccggctggg caacatgctt
     1921 cggcatggcg aatgggacca tgggagaccc tcgagccacc catgaccaaa atcccttaac
     1981 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2041 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2101 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2161 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     2221 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2281 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2341 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2401 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2461 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2521 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2581 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2641 gcggcctttt tacggttcct tgccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2701 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2761 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     2821 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     2881 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     2941 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     3001 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     3061 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     3121 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     3181 actct
//
