LOCUS       sequence-392989-        7590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dLbCas12a-D156R based adenine base editor, ecTadA8e fused to the N 
            terminal of dLbCas12a-D156R with the linker of (GSAGSAAGSGEF)x3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7590)
  AUTHORS   Cheng Y, Zhang Y, Li G, Fang H, Sretenovic S, Fan A, Li J, Xu J, Que
            Q, Qi Y
  TITLE     CRISPR-Cas12a base editors confer efficient multiplexed genome 
            editing in rice.
  JOURNAL   Plant Commun. 2023 Apr 13:100601. doi: 10.1016/j.xplc.2023.100601.
  PUBMED    37060177
REFERENCE   2  (bases 1 to 7590)
  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 Apr 13:100601. doi: 10.1016/j.xplc.2023.100601."
COMMENT     Sequence Label: dpYPQ230-D156R-ABE8
FEATURES             Location/Qualifiers
     source          1..7590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             358..1149
                     /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      1242..1830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1731..1750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1984..2001
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2160..2187)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2279..2365)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2414..2436
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2428..2445
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2429..2445
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2461..2560
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2600..2620
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3236..3256
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6962..7009
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      7040..7292
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    7337..7460
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(7477..7496)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7478..7496)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7501..7517)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7501..7517)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tggaccagcc aggacagaaa tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc
       61 acaccgtgga aacggatgaa ggcacgaacc cagtggacat aagcctgttc ggttcgtaag
      121 ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc
      181 ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt tggggtacag
      241 tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta
      301 tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt aaacattatg
      361 agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag
      421 cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt ggatggcggc
      481 ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct tgatgaaaca
      541 acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg agagagcgag
      601 attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc gtggcgttat
      661 ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct tgcaggtatc
      721 ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc aagagaacat
      781 agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc tgaacaggat
      841 ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga ctgggctggc
      901 gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa
      961 atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc ccagtatcag
     1021 cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg cttggcctcg
     1081 cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac caaggtagtc
     1141 ggcaaataac cctcgagcca cccatgacca aaatccctta acgtgagtta cgcgtcgttc
     1201 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1261 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1321 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1381 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1441 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1501 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1561 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1621 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1681 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     1741 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     1801 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     1861 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     1921 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     1981 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     2041 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     2101 agtgagcgca acgcaattaa tacgcgtacc gcgagccagg aagagtttgt agaaacgcaa
     2161 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc
     2221 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
     2281 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
     2341 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgctt
     2401 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
     2461 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
     2521 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccgatgaaac ggacagccga
     2581 cggaagcgag ttcgagtcac caaagaagaa gcggaaagtc tctgaggtgg agttttccca
     2641 cgagtactgg atgagacatg ccctgaccct ggccaagagg gcacgggatg agagggaggt
     2701 gcctgtggga gccgtgctgg tgctgaacaa tagagtgatc ggcgagggct ggaacagagc
     2761 catcggcctg cacgacccaa cagcccatgc cgaaattatg gccctgagac agggcggcct
     2821 ggtcatgcag aactacagac tgattgacgc caccctgtac gtgacattcg agccttgcgt
     2881 gatgtgcgcc ggcgccatga tccactctag gatcggccgc gtggtgtttg gcgtgaggaa
     2941 ctcaaaaaga ggcgccgcag gctccctgat gaacgtgctg aactaccccg gcatgaatca
     3001 ccgcgtcgaa attaccgagg gaatcctggc agatgaatgt gccgccctgc tgtgcgattt
     3061 ctatcggatg cctagacagg tgttcaatgc tcagaagaag gcccagagct ccatcaacgg
     3121 aagtgctggc tctgctgcgg gttcgggtga atttggttct gcgggatcgg cggcagggag
     3181 tggggagttt ggttcagctg ggtcagcggc cggctcgggt gaattcacca tggctcctaa
     3241 gaagaagcgg aaggttggta ttcacggggt gcctgcggct tcaaagctcg agaaattcac
     3301 caactgttat tcgttgagca aaacactgcg gtttaaagcg attccagtcg gcaagactca
     3361 agagaatata gacaataagc ggctgttggt ggaagatgaa aagcgcgcgg aagactacaa
     3421 aggggtgaag aagttgttgg acagatacta cctctctttt atcaatgatg tcttgcactc
     3481 aatcaaattg aagaatctga acaactacat ctccctcttc agaaagaaaa caaggacaga
     3541 aaaggagaat aaggaacttg aaaatttgga gatcaatctg aggaaagaga tcgcgaaagc
     3601 ctttaaaggc aacgaaggat acaaaagtct gttcaagaag gatataattg agacaatttt
     3661 gccagagttc ctcgatgaca aggacgagat tgcgctggtc aattcgttca acggattcac
     3721 aacagcattc acaggcttct ttcgcaatcg ggaaaatatg ttctctgagg aggcaaagtc
     3781 cacttctatt gcgttcaggt gtatcaatga gaatctcact aggtacattt ccaacatgga
     3841 tatctttgag aaggttgacg caatttttga caagcacgaa gttcaggaga ttaaggagaa
     3901 gatcctcaat tccgattatg acgttgagga cttcttcgaa ggtgagtttt ttaatttcgt
     3961 gctcactcaa gagggtatcg acgtgtataa tgcgatcatc ggtgggttcg tgactgagtc
     4021 cggtgaaaag attaagggat tgaacgagta tatcaacctt tacaaccaaa agacgaaaca
     4081 gaagctgcca aagttcaagc ctctttacaa acaggttctt tcagaccgcg agtcactctc
     4141 gttctatggg gagggctaca cttcggatga ggaagtcctg gaggtgttca ggaatactct
     4201 caataagaat tcggagattt tctcttctat aaaaaaactg gaaaagttgt ttaagaattt
     4261 tgacgaatac tctagcgccg gcatatttgt gaaaaacggc ccggccatat caacgataag
     4321 taaagatatc ttcggcgaat ggaacgtgat cagagacaaa tggaacgcgg agtatgacga
     4381 tattcacctg aagaagaagg ctgtcgtaac ggagaagtac gaggatgatc gcaggaaaag
     4441 cttcaaaaag atcggaagtt tcagcctgga acagttgcag gagtatgctg acgccgatct
     4501 tagcgtcgtc gagaagttga aggagataat catccaaaag gtcgacgaga tatataaagt
     4561 ctatggatca agtgaaaaac tgttcgacgc cgacttcgtt ttggagaagt ccctgaagaa
     4621 gaacgacgct gttgttgcca ttatgaagga tctgctcgac agcgtgaaga gtttcgagaa
     4681 ctatattaag gcttttttcg gggaggggaa ggagactaac agagatgagt ccttctacgg
     4741 agacttcgtc ctcgcgtacg atatactcct taaggtagac cacatctacg acgcaatcag
     4801 aaattacgtg acacaaaagc cgtacagcaa ggacaagttc aaactctact tccagaaccc
     4861 ccagttcatg ggcggctggg acaaggacaa ggaaacggat tacagggcta cgatcctgag
     4921 gtatggttca aaatactact tggcgattat ggacaagaag tacgccaagt gtctccagaa
     4981 gattgacaaa gacgatgtca atggcaatta tgagaagatc aactacaagc tgcttccggg
     5041 tccgaacaag atgctcccaa aggttttctt cagcaagaaa tggatggcct actataaccc
     5101 aagcgaggac atccagaaga tttataagaa cggtacgttc aagaagggcg acatgttcaa
     5161 tcttaacgac tgtcacaagc tgatcgactt cttcaaagac tcaattagcc ggtacccaaa
     5221 gtggtctaac gcctatgact tcaacttttc ggaaaccgag aagtacaagg atatagccgg
     5281 attttataga gaggtggaag agcagggcta caaggtgtca ttcgagtccg ccagcaagaa
     5341 ggaagtggac aagctcgtgg aagagggtaa gctctacatg ttccagattt ataataaaga
     5401 ctttagcgat aagagccacg ggacacctaa tctccacaca atgtatttca agctgctctt
     5461 cgacgagaat aaccacggcc aaatcaggtt gtcaggaggg gctgaactct tcatgcggcg
     5521 cgctagcctt aagaaggagg agcttgtagt ccaccctgcg aatagtccaa ttgcgaataa
     5581 gaacccggac aatcctaaaa agactacaac attgagctac gacgtgtaca aggataagag
     5641 gttttccgag gatcagtacg agctccacat cccgattgcg atcaacaagt gcccaaagaa
     5701 tattttcaag ataaacacag aggtgcgtgt actcctgaag catgacgaca atccttacgt
     5761 cattgggatt gcccggggcg agaggaacct cctctatatt gtggtggtgg acgggaaggg
     5821 gaacatagtc gaacagtact cccttaacga aataattaac aatttcaacg gcatccgtat
     5881 caagaccgac taccattcgt tgctggacaa gaaggagaag gagagatttg aggcgcggca
     5941 aaattggaca agtatcgaga acatcaagga actcaaagca ggttatatct ctcaagttgt
     6001 gcataagata tgcgagctgg ttgagaagta tgacgcagtg atcgctcttg aggacctcaa
     6061 ctcgggcttt aagaattcta gagttaaagt ggagaagcag gtctatcaaa agttcgagaa
     6121 gatgcttata gataagctca actacatggt cgataagaaa tcgaacccat gtgccaccgg
     6181 cggcgcactc aaaggttacc aaataacaaa caaattcgag tccttcaaat cgatgagtac
     6241 tcagaatggg ttcatatttt atataccggc gtggcttacg tctaagatcg acccgtcaac
     6301 tggttttgtc aacctgttga agacgaaata cacgtccatt gccgattcga aaaagttcat
     6361 atctagtttt gatcgtatta tgtacgtccc agaggaagat cttttcgagt ttgctctcga
     6421 ctacaaaaac ttttcgcgga ccgatgcgga ttacattaaa aaatggaaac tctattcgta
     6481 cggcaacaga atcaggattt ttcgcaaccc taagaagaat aacgtctttg attgggagga
     6541 agtttgcttg actagcgcgt acaaggagct ctttaataag tatggcatta actaccaaca
     6601 gggtgatatc agagcactgc tttgcgaaca atctgacaag gctttctact catccttcat
     6661 ggctttgatg agcctgatgc tccagatgag aaattcaatt acaggcagaa ccgacgtgga
     6721 tttcttgatc tccccggtta aaaattctga tggcatcttt tacgatagca ggaactatga
     6781 agcgcaagag aatgcgattc tgccaaaaaa tgcagacgcc aacggtgcct ataacatcgc
     6841 caggaaagtc ctgtgggcga tcggccagtt caaaaaggcc gaagacgaaa aattggacaa
     6901 ggtcaaaatc gctatcagca acaaagagtg gctggagtat gctcagacat ccgtaaagca
     6961 taagcgtcct gctgccacca aaaaggccgg acaggctaag aaaaagaagt gagacgacta
     7021 gtggcggccg ccgacgtccg atcgttcaaa catttggcaa taaagtttct taagattgaa
     7081 tcctgttgcc ggtcttgcga tgattatcat ataatttctg ttgaattacg ttaagcatgt
     7141 aataattaac atgtaatgca tgacgttatt tatgagatgg gtttttatga ttagagtccc
     7201 gcaattatac atttaatacg cgatagaaaa caaaatatag cgcgcaaact aggataaatt
     7261 atcgcgcgcg gtgtcatcta tgttactaga tcgggaattg atcccccctc gacagcttcc
     7321 ggaaagggcg aattcgcaac tttgtataca aaagttgaac gagaaacgta aaatgatata
     7381 aatatcaata tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca
     7441 caacatatcc agtcactatg ccatccagct gatatcccct atagtgagtc gtattacatg
     7501 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     7561 caagataaaa atatatcatc atgcctcctc
//
