LOCUS       sequence-422150-        4645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Modular Unit carrying pegRNA and ngRNA scaffolds flanked by attL3 
            and attL2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4645)
  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 4645)
  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: pCR8-attL3-pegRNA-attL2
FEATURES             Location/Qualifiers
     source          1..4645
                     /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..2793
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     ncRNA           4040..4107
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     misc_RNA        4213..4288
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           4298..4365
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     terminator      4397..4645
                     /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 cgggcccaaa
     2701 taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat gagcaatgct
     2761 tttttataat gccaactttg tataataaag ttgccgaatt atggagtcaa agattcaaat
     2821 agaggaccta acagaactcg ccgtaaagac tggcgaacag ttcatacaga gtctcttacg
     2881 actcaatgac aagaagaaaa tcttcgtcaa catggtggag cacgacacac ttgtctactc
     2941 caaaaatatc aaagatacag tctcagaaga ccaaagggca attgagactt ttcaacaaag
     3001 ggtaatatcc ggaaacctcc tcggattcca ttgcccagct atctgtcact ttattgtgaa
     3061 gatagtggaa aaggaaggtg gctcctacaa atgccatcat tgcgataaag gaaaggccat
     3121 cgttgaagat gcctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat
     3181 cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgattggca
     3241 gacatactgt cccacaaatg aagatggaat ctgtaaaaga aaacgcgtga aataatgcgt
     3301 ctgacaaagg ttaggtcggc tgcctttaat caataccaaa gtggtcccta ccacgatgga
     3361 aaaactgtgc agtcggtttg gctttttctg acgaacaaat aagattcgtg gccgacaggt
     3421 gggggtccac catgtgaagg catcttcaga ctccaataat ggagcaatga cgtaagggct
     3481 tacgaaataa gtaagggtag tttgggaaat gtccactcac ccgtcagtct ataaatactt
     3541 agcccctccc tcattgttaa gggagcaaaa tctcagagag atagtcctag agagagaaag
     3601 agagcaagta gcctagaagt agtcaaggcg gcgaagtatt caggcacgtg gccaggaaga
     3661 agaaaagcca agacgacgaa aacaggtaag agctaagcat ctagaaagtt gaaaacaatc
     3721 ttcaaaagtc ccacatcgct tagataagaa aacgaagctg agtttatata cagctagagt
     3781 cgaagtagtg attgaacaaa gcaccagtgg tctagtggta gaatagtacc ctgccacggt
     3841 acagacccgg gttcgattcc cggctggtgc aagagacgag tcacgtctca gtttaagagc
     3901 tatgctggaa acagcatagc aagtttaaat aaggctagtc cgttatcaac ttgaaaaagt
     3961 ggcaccgagt cggtgcagag accacaggat ccttggtctc acgcggttct atctagttac
     4021 gcgttaaacc aactagaaag gccggcatgg tcccagcctc ctcgctggcg ccggctgggc
     4081 aacatgcttc ggcatggcga atgggacaac aacaaatcag agtggcgcag cggaagcgtg
     4141 gtgggcccat aacccacagg tcccaggatc gaaacctggc tctgataaga gaccacagga
     4201 tccttggtct cagttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
     4261 acttgaaaaa gtggcaccga gtcggtgctt tttttttggc cggcatggtc ccagcctcct
     4321 cgctggcgcc ggctgggcaa catgcttcgg catggcgaat gggacttttt ttttgatatc
     4381 tccggggcta attgaatatg aagatgaaga tgaaatattt ggtgtgtcaa ataaaaagct
     4441 ggtgtgctta agtttgtgtt tttttcttgg cttgttgtgt tatgaatttg tggctttttc
     4501 taatattaaa tgaatgtaag atctcattat aatgaataaa caaatgtttc tataatccat
     4561 tgtgaatgtt ttgttggatc tcttctgcag catataacta ctgtatgtgc tatggtatgg
     4621 actatggaat atgattaaag ataag
//
