LOCUS       sequence-422146-        4649 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Modular Unit carrying pegRNA and ngRNA scaffolds flanked by attL1 
            and attL2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4649)
  AUTHORS   Gupta A, Liu B, Raza S, Chen QJ, Yang B
  TITLE     Modularly assembled multiplex prime editors for simultaneous editing
            of agronomically important genes in rice.
  JOURNAL   Plant Commun. 2023 Oct 26:100741. doi: 10.1016/j.xplc.2023.100741.
  PUBMED    37897041
REFERENCE   2  (bases 1 to 4649)
  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. 2023 Oct 26:100741. doi: 10.1016/j.xplc.2023.100741."
COMMENT     Sequence Label: pAG79-attL1-pegRNA-attL2
FEATURES             Location/Qualifiers
     source          1..4649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(8..107)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(124..143)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(125..143)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(148..164)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(148..164)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             595..1386
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1479..2067
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1968..1987
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2221..2238
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2397..2424)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2516..2602)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2651..2673
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2665..2682
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2666..2682
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2698..2797
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     ncRNA           4044..4111
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     misc_RNA        4217..4292
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           4302..4369
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     terminator      4401..4649
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
ORIGIN
        1 gaattcgacc cagctttctt gtacaaagtt ggcattataa aaaataattg ctcatcaatt
       61 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat
      121 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
      181 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg cctcctctag
      241 accagccagg acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca
      301 ccgtggaaac ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg
      361 taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt
      421 ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct
      481 atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg
      541 agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg
      601 gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc
      661 catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg
      721 aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg
      781 cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt
      841 ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca
      901 gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc
      961 gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc
     1021 gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta
     1081 tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat
     1141 gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc
     1201 gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc
     1261 gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc
     1321 gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc
     1381 aaataaccct cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac
     1441 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1501 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1561 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1621 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1681 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1741 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1801 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1861 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     1921 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     1981 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2041 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2101 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2161 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     2221 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     2281 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     2341 gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa
     2401 ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc
     2461 ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg
     2521 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg
     2581 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca
     2641 tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct cgggccccaa
     2701 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga tgagcaatgc
     2761 ttttttataa tgccaacttt gtacaaaaaa gcaggctccg aattatggag tcaaagattc
     2821 aaatagagga cctaacagaa ctcgccgtaa agactggcga acagttcata cagagtctct
     2881 tacgactcaa tgacaagaag aaaatcttcg tcaacatggt ggagcacgac acacttgtct
     2941 actccaaaaa tatcaaagat acagtctcag aagaccaaag ggcaattgag acttttcaac
     3001 aaagggtaat atccggaaac ctcctcggat tccattgccc agctatctgt cactttattg
     3061 tgaagatagt ggaaaaggaa ggtggctcct acaaatgcca tcattgcgat aaaggaaagg
     3121 ccatcgttga agatgcctct gccgacagtg gtcccaaaga tggaccccca cccacgagga
     3181 gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgatt
     3241 ggcagacata ctgtcccaca aatgaagatg gaatctgtaa aagaaaacgc gtgaaataat
     3301 gcgtctgaca aaggttaggt cggctgcctt taatcaatac caaagtggtc cctaccacga
     3361 tggaaaaact gtgcagtcgg tttggctttt tctgacgaac aaataagatt cgtggccgac
     3421 aggtgggggt ccaccatgtg aaggcatctt cagactccaa taatggagca atgacgtaag
     3481 ggcttacgaa ataagtaagg gtagtttggg aaatgtccac tcacccgtca gtctataaat
     3541 acttagcccc tccctcattg ttaagggagc aaaatctcag agagatagtc ctagagagag
     3601 aaagagagca agtagcctag aagtagtcaa ggcggcgaag tattcaggca cgtggccagg
     3661 aagaagaaaa gccaagacga cgaaaacagg taagagctaa gcatctagaa agttgaaaac
     3721 aatcttcaaa agtcccacat cgcttagata agaaaacgaa gctgagttta tatacagcta
     3781 gagtcgaagt agtgattgaa caaagcacca gtggtctagt ggtagaatag taccctgcca
     3841 cggtacagac ccgggttcga ttcccggctg gtgcaagaga cgagtcacgt ctcagtttaa
     3901 gagctatgct ggaaacagca tagcaagttt aaataaggct agtccgttat caacttgaaa
     3961 aagtggcacc gagtcggtgc agagaccaca ggatccttgg tctcacgcgg ttctatctag
     4021 ttacgcgtta aaccaactag aaaggccggc atggtcccag cctcctcgct ggcgccggct
     4081 gggcaacatg cttcggcatg gcgaatggga caacaacaaa tcagagtggc gcagcggaag
     4141 cgtggtgggc ccataaccca caggtcccag gatcgaaacc tggctctgat aagagaccac
     4201 aggatccttg gtctcagttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
     4261 atcaacttga aaaagtggca ccgagtcggt gctttttttt tggccggcat ggtcccagcc
     4321 tcctcgctgg cgccggctgg gcaacatgct tcggcatggc gaatgggact ttttttttga
     4381 tatctccggg gctaattgaa tatgaagatg aagatgaaat atttggtgtg tcaaataaaa
     4441 agctggtgtg cttaagtttg tgtttttttc ttggcttgtt gtgttatgaa tttgtggctt
     4501 tttctaatat taaatgaatg taagatctca ttataatgaa taaacaaatg tttctataat
     4561 ccattgtgaa tgttttgttg gatctcttct gcagcatata actactgtat gtgctatggt
     4621 atggactatg gaatatgatt aaagataag
//
