LOCUS       sequence-375104-        3185 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Golden Gate entry vector to clone the 3rd 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 Apr 14, 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: pYPQ133-RZ-Cas12j2
FEATURES             Location/Qualifiers
     source          1..3185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(45..131)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(355..360,361..377,378..383)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             431..1621
                     /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(783..802)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(1795..1814)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1796..1814)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1819..1835)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1819..1835)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     ncRNA           1930..1966
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     ncRNA           2024..2091
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     rep_origin      2193..2781
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2682..2701
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3111..3138)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg
       61 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt
      121 cgttttattt gatgcctggc agttccctac tctcgcgttc tgcattaatg aatcggccaa
      181 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
      241 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
      301 ttatccacag aatcagggga taacgcagga aagaacatga attaattctc atgtttgaca
      361 gcttatcatc gattagcttt aatgcggtag tttatcacag ttaaattgct aacgcagtca
      421 ggcaccgtgt atgaaatcta acaatgcgct catcgtcatc ctcggcaccg tcaccctgga
      481 tgctgtaggc ataggcttgg ttatgccggt actgccgggc ctcttgcggg atatcgtcca
      541 ttccgacagc atcgccagtc actatggcgt gctgctagcg ctatatgcgt tgatgcaatt
      601 tctatgcgca cccgttctcg gagcactgtc cgaccgcttt ggccgccgcc cagtcctgct
      661 cgcttcgcta cttggagcca ctatcgacta cgcgatcatg gcgaccacac ccgtcctgtg
      721 gattctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg
      781 cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag
      841 cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat
      901 ctccttacat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg
      961 ctgcttccta atgcaggagt cgcataaggg agagcgccga cccatgccct tgagagcctt
     1021 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     1081 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg
     1141 cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat
     1201 cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa
     1261 gcaggccatt atcgccggca tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc
     1321 gacgcgaggc tggatggcct tccccattat gattcttctc gcttccggcg gcatcgggat
     1381 gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg gacagcttca
     1441 aggatcgctc gcggctctta ccagcctaac ttcgatcatt ggaccgctga tcgtcacggc
     1501 gatttatgcc gcctcggcga gcacatggaa cgggttggca tggattgtag gcgccgccct
     1561 ataccttgtc tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca cctcgacctg
     1621 aatggaagcc ggcggcacct cgctaacgga ttcaccactc caagaattgg agccaatcaa
     1681 ttcttgcgga gaactgtgaa tgcgcaaacc aacccttgat cggggaagaa cagtatgtcg
     1741 agctattttt tgacttactg gggatcaagc ctgattggga gaaaataaaa tatcccctat
     1801 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     1861 ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctcta gaggtctcag
     1921 gactcgttgc tgatgagtcc gtgaggacga aacgagtaag ctcgtccaac gattgcccct
     1981 cacgagggga cgagacggag ctcagtctga ccgcggcgtc tctggccggc atggtcccag
     2041 cctcctcgct ggcgccggct gggcaacatg cttcggcatg gcgaatggga cccagggaga
     2101 ccctcgagcc acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     2161 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2221 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2281 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2341 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2401 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2461 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2521 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2581 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2641 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2701 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2761 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     2821 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     2881 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     2941 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     3001 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     3061 aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat
     3121 ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc
     3181 gccac
//
