LOCUS       sequence-324566-        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"
     misc_RNA        681..756
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      845..1093
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     protein_bind    complement(1191..1290)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1307..1326)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1308..1326)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1331..1347)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1331..1347)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1778..2569
                     /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      2662..3250
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3151..3170
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3404..3421
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3580..3607)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3699..3785)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3834..3856
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3848..3865
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3849..3865
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    join(3881..3898,1..82)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 tgactgatag tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca
       61 actttgtaca aaaaagcagg ctccggaatt cacctgccag gggacctggc agacatactg
      121 tcccacaaat gaagatggaa tctgtaaaag aaaacgcgtg aaataatgcg tctgacaaag
      181 gttaggtcgg ctgcctttaa tcaataccaa agtggtccct accacgatgg aaaaactgtg
      241 cagtcggttt ggctttttct gacgaacaaa taagattcgt ggccgacagg tgggggtcca
      301 ccatgtgaag gcatcttcag actccaataa tggagcaatg acgtaagggc ttacgaaata
      361 agtaagggta gtttgggaaa tgtccactca cccgtcagtc tataaatact tagcccctcc
      421 ctcattgtta agggagcaaa atctcagaga gatagtccta gagagagaaa gagagcaagt
      481 agcctagaag tagtcaaggc ggcgaagtat tcaggcacgt ggccaggaag aagaaaagcc
      541 aagacgacga aaacaggtaa gagctaagct tcgcatgcag aacaaagcac cagtggtcta
      601 gtggtagaat agtaccctgc cacggtacag acccgggttc gattcccggc tggtgcaaga
      661 gaccaagacg tcaggtctca gttttagagc tagaaatagc aagttaaaat aaggctagtc
      721 cgttatcaac ttgaaaaagt ggcaccgagt cggtgcaaca aagcaccagt ggtctagtgg
      781 tagaatagta ccctgccacg gtacagaccc gggttcgatt cccggctggt gcagtcgatc
      841 aagatatgaa gatgaagatg aaatatttgg tgtgtcaaat aaaaagcttg tgtgcttaag
      901 tttgtgtttt tttcttggct tgttgtgtta tgaatttgtg gctttttcta atattaaatg
      961 aatgtaagat cacattataa tgaataaaca aatgtttcta taatccattg tgaatgtttt
     1021 gttggatctc ttctgcagca tataactact gtatgtgcta tggtatggac tatggaatat
     1081 gattaaagat aagaactcgc gcgtagtcct cggtaatatc gcagaacaaa agtacctgat
     1141 atcgagtgta cttcaagtca caccggcgag tgactagcag gtggaattcg acccagcttt
     1201 cttgtacaaa gttggcatta taaaaaataa ttgctcatca atttgttgca acgaacaggt
     1261 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     1321 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     1381 ttacattgca caagataaaa atatatcatc atgcctcctc tagaccagcc aggacagaaa
     1441 tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa
     1501 ggcacgaacc cagtggacat aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat
     1561 gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc
     1621 ggttttcatg gcttgttatg actgtttttt tggggtacag tctatgcctc gggcatccaa
     1681 gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac
     1741 gcagcagggc agtcgcccta aaacaaagtt aaacattatg agggaagcgg tgatcgccga
     1801 agtatcgact caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt
     1861 gctggccgta catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat
     1921 tgatttgctg gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa
     1981 cgaccttttg gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt
     2041 caccattgtt gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca
     2101 atttggagaa tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga
     2161 cattgatctg gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc
     2221 agcggcggag gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga
     2281 aaccttaacg ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct
     2341 tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc
     2401 tgccgactgg gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag
     2461 acaggcttat cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga
     2521 atttgtccac tacgtgaaag gcgagatcac caaggtagtc ggcaaataac cctcgagcca
     2581 cccatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     2641 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2701 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2761 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     2821 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2881 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2941 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3001 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3061 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3121 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3181 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3241 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3301 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3361 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3421 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     3481 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     3541 tacgcgtacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg
     3601 gccttctgct tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg
     3661 ggccgttgct tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg
     3721 ttcaccgaca aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt
     3781 atttgatgcc tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc
     3841 acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc caaataatga ttttattt
//
