LOCUS       sequence-375103-        3185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate entry vector to clone the 2nd 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: pYPQ132-RZ-Cas12j2
FEATURES             Location/Qualifiers
     source          1..3185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(203..208,209..225,226..231)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             279..1469
                     /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(631..650)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(1643..1662)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1644..1662)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1667..1683)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1667..1683)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     ncRNA           1778..1814
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     ncRNA           1872..1939
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     rep_origin      2041..2629
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2530..2549
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2959..2986)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3078..3164)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tcgcgttctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
       61 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
      121 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      181 gaacatgaat taattctcat gtttgacagc ttatcatcga ttagctttaa tgcggtagtt
      241 tatcacagtt aaattgctaa cgcagtcagg caccgtgtat gaaatctaac aatgcgctca
      301 tcgtcatcct cggcaccgtc accctggatg ctgtaggcat aggcttggtt atgccggtac
      361 tgccgggcct cttgcgggat atcgtccatt ccgacagcat cgccagtcac tatggcgtgc
      421 tgctagcgct atatgcgttg atgcaatttc tatgcgcacc cgttctcgga gcactgtccg
      481 accgctttgg ccgccgccca gtcctgctcg cttcgctact tggagccact atcgactacg
      541 cgatcatggc gaccacaccc gtcctgtgga ttctctacgc cggacgcatc gtggccggca
      601 tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag
      661 atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc
      721 ccgtggccgg gggactgttg ggcgccatct ccttacatgc accattcctt gcggcggcgg
      781 tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag
      841 agcgccgacc catgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
      901 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
      961 tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga
     1021 tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg
     1081 gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg
     1141 cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga
     1201 ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg
     1261 tagatgacga ccatcaggga cagcttcaag gatcgctcgc ggctcttacc agcctaactt
     1321 cgatcattgg accgctgatc gtcacggcga tttatgccgc ctcggcgagc acatggaacg
     1381 ggttggcatg gattgtaggc gccgccctat accttgtctg cctccccgcg ttgcgtcgcg
     1441 gtgcatggag ccgggccacc tcgacctgaa tggaagccgg cggcacctcg ctaacggatt
     1501 caccactcca agaattggag ccaatcaatt cttgcggaga actgtgaatg cgcaaaccaa
     1561 cccttgatcg gggaagaaca gtatgtcgag ctattttttg acttactggg gatcaagcct
     1621 gattgggaga aaataaaata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     1681 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     1741 atcatcatgc ctcctctaga ggtctcccat gtcgttgctg atgagtccgt gaggacgaaa
     1801 cgagtaagct cgtccaacga ttgcccctca cgaggggacg agacggagct cagtctgacc
     1861 gcggcgtctc tggccggcat ggtcccagcc tcctcgctgg cgccggctgg gcaacatgct
     1921 tcggcatggc gaatgggacg gaccgagacc ctcgagccac ccatgaccaa aatcccttaa
     1981 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2041 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2101 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2161 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     2221 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2281 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2341 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2401 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2461 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2521 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2581 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2641 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2701 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     2761 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     2821 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     2881 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga
     2941 agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc
     3001 ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc
     3061 aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
     3121 aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc
     3181 tactc
//
