LOCUS       sequence-316059-        3898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector for gRNA insertion in plant prime editing expression 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3898)
  AUTHORS   Wang L, Kaya HB, Zhang N, Rai R, Willmann MR, Carpenter SCD, Read 
            AC, Martin F, Fei Z, Leach JE, Martin GB, Bogdanove AJ
  TITLE     Spelling Changes and Fluorescent Tagging With Prime Editing Vectors 
            for Plants.
  JOURNAL   Front Genome Ed. 2021 Mar 4;3:617553. doi: 
            10.3389/fgeed.2021.617553. eCollection 2021.
  PUBMED    34713247
REFERENCE   2  (bases 1 to 3898)
  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: "Front 
            Genome Ed."; date: "2021-03-4"; volume: "3:617553. doi"; pages: " 
            10.3389/fgeed.2021.617553. eCollection 2021"
COMMENT     Sequence Label: pPEG
FEATURES             Location/Qualifiers
     source          1..3898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_RNA        731..806
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      895..1143
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     protein_bind    complement(1241..1340)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1357..1376)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1358..1376)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1381..1397)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1381..1397)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1828..2619
                     /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      2712..3300
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3201..3220
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3454..3471
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3630..3657)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3749..3835)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3884..3898,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3898,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagcagg ctccggaatt cacctgccag gggacctggc agacatactg tcccacaaat
      181 gaagatggaa tctgtaaaag aaaacgcgtg aaataatgcg tctgacaaag gttaggtcgg
      241 ctgcctttaa tcaataccaa agtggtccct accacgatgg aaaaactgtg cagtcggttt
      301 ggctttttct gacgaacaaa taagattcgt ggccgacagg tgggggtcca ccatgtgaag
      361 gcatcttcag actccaataa tggagcaatg acgtaagggc ttacgaaata agtaagggta
      421 gtttgggaaa tgtccactca cccgtcagtc tataaatact tagcccctcc ctcattgtta
      481 agggagcaaa atctcagaga gatagtccta gagagagaaa gagagcaagt agcctagaag
      541 tagtcaaggc ggcgaagtat tcaggcacgt ggccaggaag aagaaaagcc aagacgacga
      601 aaacaggtaa gagctaagct tcgcatgcag aacaaagcac cagtggtcta gtggtagaat
      661 agtaccctgc cacggtacag acccgggttc gattcccggc tggtgcaaga gaccaagacg
      721 tcaggtctca gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac
      781 ttgaaaaagt ggcaccgagt cggtgcaaca aagcaccagt ggtctagtgg tagaatagta
      841 ccctgccacg gtacagaccc gggttcgatt cccggctggt gcagtcgatc aagatatgaa
      901 gatgaagatg aaatatttgg tgtgtcaaat aaaaagcttg tgtgcttaag tttgtgtttt
      961 tttcttggct tgttgtgtta tgaatttgtg gctttttcta atattaaatg aatgtaagat
     1021 cacattataa tgaataaaca aatgtttcta taatccattg tgaatgtttt gttggatctc
     1081 ttctgcagca tataactact gtatgtgcta tggtatggac tatggaatat gattaaagat
     1141 aagaactcgc gcgtagtcct cggtaatatc gcagaacaaa agtacctgat atcgagtgta
     1201 cttcaagtca caccggcgag tgactagcag gtggaattcg acccagcttt cttgtacaaa
     1261 gttggcatta taaaaaataa ttgctcatca atttgttgca acgaacaggt cactatcagt
     1321 caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg
     1381 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     1441 caagataaaa atatatcatc atgcctcctc tagaccagcc aggacagaaa tgcctcgact
     1501 tcgctgctgc ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc
     1561 cagtggacat aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc
     1621 aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg
     1681 gcttgttatg actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg
     1741 cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc
     1801 agtcgcccta aaacaaagtt aaacatcatg agggaagcgg tgatcgccga agtatcgact
     1861 caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta
     1921 catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg
     1981 gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg
     2041 gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt
     2101 gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa
     2161 tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg
     2221 gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag
     2281 gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg
     2341 ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc
     2401 cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg
     2461 gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat
     2521 cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac
     2581 tacgtgaaag gcgagatcac caaggtagtc ggcaaataac cctcgagcca cccatgacca
     2641 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     2701 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     2761 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2821 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     2881 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     2941 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3001 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3061 gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg
     3121 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3181 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3241 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3301 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3361 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     3421 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     3481 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     3541 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
     3601 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
     3661 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
     3721 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
     3781 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
     3841 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgtt
//
