LOCUS       sequence-298972-        3406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway compatible prime editing gRNA empty backbone vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3406)
  AUTHORS   Tang X, Sretenovic S, Ren Q, Jia X, Li M, Fan T, Yin D, Xiang S, Guo
            Y, Liu L, Zheng X, Qi Y, Zhang Y
  TITLE     Plant Prime Editors Enable Precise Gene Editing in Rice Cells.
  JOURNAL   Mol Plant. 2020 Mar 25. pii: S1674-2052(20)30072-1. doi: 
            10.1016/j.molp.2020.03.010.
  PUBMED    32222487
REFERENCE   2  (bases 1 to 3406)
  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: "Mol Plant. 
            2020 Mar 25. pii: S1674-2052(20)30072-1. doi: 
            10.1016/j.molp.2020.03.010."
COMMENT     Sequence Label: pYPQ141D-peg
FEATURES             Location/Qualifiers
     source          1..3406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..398
                     /label=OsU3 promoter
                     /note="Oryza sativa (rice) snRNA U3 promoter"
     misc_RNA        431..506
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(606..705)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(722..741)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(723..741)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(746..762)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(746..762)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1193..1984
                     /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      2077..2665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2566..2585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2819..2836
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2995..3022)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3114..3200)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3249..3271
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3263..3280
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3264..3280
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3295..3390
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ctagataacg caggatccta aagggatctt taaacatacg aacagatcac ttaaagttct
       61 tctgaagcaa cttaaagtta tcaggcatgc atggatcttg gaggaatcag atgtgcagtc
      121 agggaccata gcacaggaca ggcgtcttct actggtgcta ccagcaaatg ctggaagccg
      181 ggaacactgg gtacgttgga aaccacgtga tgtggagtaa gataaactgt aggagaaaag
      241 catttcgtag tgggccatga agcctttcag gacatgtatt gcagtatggg ccggcccatt
      301 acgcaattgg acgacaacaa agactagtat tagtaccacc tcggctatcc acatagatca
      361 aagctggttt aaaagagttg tgcagatgat ccgtggcaga gacgagatct agtctgagtc
      421 gaccgtctct gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac
      481 ttgaaaaagt ggcaccgagt cggtgcggag accgacgtca attgctagcg gtctcatttt
      541 tttctgcaga tatccatcac actggcggcc gctcgaggtc gagggtatcg ataagcttga
      601 attcgaccca gctttcttgt acaaagttgg cattataaaa aataattgct catcaatttg
      661 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
      721 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
      781 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc tcctctggac
      841 cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc
      901 gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta
      961 atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg
     1021 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat
     1081 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
     1141 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga
     1201 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
     1261 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
     1321 gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
     1381 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
     1441 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
     1501 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
     1561 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
     1621 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
     1681 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
     1741 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
     1801 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
     1861 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
     1921 agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa
     1981 ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     2041 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2101 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2161 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2221 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2281 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2341 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2401 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2461 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2521 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2581 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     2641 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     2701 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     2761 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     2821 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     2881 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     2941 gcgcaacgca attaatacgc gtaccgcgag ccaggaagag tttgtagaaa cgcaaaaagg
     3001 ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct
     3061 gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc
     3121 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac
     3181 tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa cgcttgcatg
     3241 gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggcccaataa
     3301 tgattttatt ttgactgata gtgacctgtt cgttgcaaca aattgatgag caatgctttt
     3361 ttataatgcc aactttgtat acaaaagttg ccccatggcg ttccct
//
