LOCUS       sequence-422148-        4692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Modular Unit carrying pegRNA and ngRNA scaffolds flanked by attL1 
            and attR3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4692)
  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 4692)
  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-attL1-pegRNA-attR3
FEATURES             Location/Qualifiers
     source          1..4692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      882..1470
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1371..1390
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1624..1641
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1800..1827)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1919..2005)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2054..2076
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2068..2085
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2069..2085
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2101..2200
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     ncRNA           3447..3514
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     misc_RNA        3620..3695
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           3705..3772
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     terminator      3804..4052
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     protein_bind    4060..4183
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4219..4238)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4220..4238)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4243..4259)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4243..4259)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(4690..4692,1..789)
                     /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"
ORIGIN
        1 agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag
       61 cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt ggatggcggc
      121 ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct tgatgaaaca
      181 acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg agagagcgag
      241 attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc gtggcgttat
      301 ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct tgcaggtatc
      361 ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc aagagaacat
      421 agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc tgaacaggat
      481 ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga ctgggctggc
      541 gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa
      601 atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc ccagtatcag
      661 cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg cttggcctcg
      721 cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac caaggtagtc
      781 ggcaaataac cctcgagcca cccatgacca aaatccctta acgtgagtta cgcgtcgttc
      841 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
      901 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
      961 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1021 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1081 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1141 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1201 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1261 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1321 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     1381 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     1441 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     1501 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     1561 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     1621 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     1681 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     1741 agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa
     1801 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc
     1861 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
     1921 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
     1981 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta
     2041 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
     2101 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
     2161 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccgaattatg gagtcaaaga
     2221 ttcaaataga ggacctaaca gaactcgccg taaagactgg cgaacagttc atacagagtc
     2281 tcttacgact caatgacaag aagaaaatct tcgtcaacat ggtggagcac gacacacttg
     2341 tctactccaa aaatatcaaa gatacagtct cagaagacca aagggcaatt gagacttttc
     2401 aacaaagggt aatatccgga aacctcctcg gattccattg cccagctatc tgtcacttta
     2461 ttgtgaagat agtggaaaag gaaggtggct cctacaaatg ccatcattgc gataaaggaa
     2521 aggccatcgt tgaagatgcc tctgccgaca gtggtcccaa agatggaccc ccacccacga
     2581 ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg
     2641 attggcagac atactgtccc acaaatgaag atggaatctg taaaagaaaa cgcgtgaaat
     2701 aatgcgtctg acaaaggtta ggtcggctgc ctttaatcaa taccaaagtg gtccctacca
     2761 cgatggaaaa actgtgcagt cggtttggct ttttctgacg aacaaataag attcgtggcc
     2821 gacaggtggg ggtccaccat gtgaaggcat cttcagactc caataatgga gcaatgacgt
     2881 aagggcttac gaaataagta agggtagttt gggaaatgtc cactcacccg tcagtctata
     2941 aatacttagc ccctccctca ttgttaaggg agcaaaatct cagagagata gtcctagaga
     3001 gagaaagaga gcaagtagcc tagaagtagt caaggcggcg aagtattcag gcacgtggcc
     3061 aggaagaaga aaagccaaga cgacgaaaac aggtaagagc taagcatcta gaaagttgaa
     3121 aacaatcttc aaaagtccca catcgcttag ataagaaaac gaagctgagt ttatatacag
     3181 ctagagtcga agtagtgatt gaacaaagca ccagtggtct agtggtagaa tagtaccctg
     3241 ccacggtaca gacccgggtt cgattcccgg ctggtgcaag agacgagtca cgtctcagtt
     3301 taagagctat gctggaaaca gcatagcaag tttaaataag gctagtccgt tatcaacttg
     3361 aaaaagtggc accgagtcgg tgcagagacc acaggatcct tggtctcacg cggttctatc
     3421 tagttacgcg ttaaaccaac tagaaaggcc ggcatggtcc cagcctcctc gctggcgccg
     3481 gctgggcaac atgcttcggc atggcgaatg ggacaacaac aaatcagagt ggcgcagcgg
     3541 aagcgtggtg ggcccataac ccacaggtcc caggatcgaa acctggctct gataagagac
     3601 cacaggatcc ttggtctcag ttttagagct agaaatagca agttaaaata aggctagtcc
     3661 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttttggccgg catggtccca
     3721 gcctcctcgc tggcgccggc tgggcaacat gcttcggcat ggcgaatggg actttttttt
     3781 tgatatctcc ggggctaatt gaatatgaag atgaagatga aatatttggt gtgtcaaata
     3841 aaaagctggt gtgcttaagt ttgtgttttt ttcttggctt gttgtgttat gaatttgtgg
     3901 ctttttctaa tattaaatga atgtaagatc tcattataat gaataaacaa atgtttctat
     3961 aatccattgt gaatgttttg ttggatctct tctgcagcat ataactactg tatgtgctat
     4021 ggtatggact atggaatatg attaaagata aggaattcgc aactttgtat aataaagttg
     4081 aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg cataaaaaac
     4141 agactacata atactgtaaa acacaacata tccagtcact atgaaaataa aatcattatt
     4201 tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg
     4261 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
     4321 tcatgcctcc tctagaccag ccaggacaga aatgcctcga cttcgctgct acccaaggtt
     4381 gccgggtgac gcacaccgtg gaaacggatg aaggcacgaa cccagtggac ataagcctgt
     4441 tcggttcgta agctgtaatg caagtagcgt atgcgctcac gcaactggtc cagaaccttg
     4501 accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt
     4561 tttggggtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga
     4621 tgtttgatgt tatggagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag
     4681 ttaaacatta tg
//
