LOCUS       sequence-392938-        7572 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 (GGGGS)x6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7572)
  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 7572)
  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-ABE4
FEATURES             Location/Qualifiers
     source          1..7572
                     /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             3218..3238
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6944..6991
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      7022..7274
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    7319..7442
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(7459..7478)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7460..7478)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7483..7499)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7483..7499)
                     /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 gggcgggggc tcaggcgggg gcgggagcgg cggcgggggc tctgggggcg gcggcagcgg
     3181 cgggggcggc agcgggggcg gcgggtcgac catggctcct aagaagaagc ggaaggttgg
     3241 tattcacggg gtgcctgcgg cttcaaagct cgagaaattc accaactgtt attcgttgag
     3301 caaaacactg cggtttaaag cgattccagt cggcaagact caagagaata tagacaataa
     3361 gcggctgttg gtggaagatg aaaagcgcgc ggaagactac aaaggggtga agaagttgtt
     3421 ggacagatac tacctctctt ttatcaatga tgtcttgcac tcaatcaaat tgaagaatct
     3481 gaacaactac atctccctct tcagaaagaa aacaaggaca gaaaaggaga ataaggaact
     3541 tgaaaatttg gagatcaatc tgaggaaaga gatcgcgaaa gcctttaaag gcaacgaagg
     3601 atacaaaagt ctgttcaaga aggatataat tgagacaatt ttgccagagt tcctcgatga
     3661 caaggacgag attgcgctgg tcaattcgtt caacggattc acaacagcat tcacaggctt
     3721 ctttcgcaat cgggaaaata tgttctctga ggaggcaaag tccacttcta ttgcgttcag
     3781 gtgtatcaat gagaatctca ctaggtacat ttccaacatg gatatctttg agaaggttga
     3841 cgcaattttt gacaagcacg aagttcagga gattaaggag aagatcctca attccgatta
     3901 tgacgttgag gacttcttcg aaggtgagtt ttttaatttc gtgctcactc aagagggtat
     3961 cgacgtgtat aatgcgatca tcggtgggtt cgtgactgag tccggtgaaa agattaaggg
     4021 attgaacgag tatatcaacc tttacaacca aaagacgaaa cagaagctgc caaagttcaa
     4081 gcctctttac aaacaggttc tttcagaccg cgagtcactc tcgttctatg gggagggcta
     4141 cacttcggat gaggaagtcc tggaggtgtt caggaatact ctcaataaga attcggagat
     4201 tttctcttct ataaaaaaac tggaaaagtt gtttaagaat tttgacgaat actctagcgc
     4261 cggcatattt gtgaaaaacg gcccggccat atcaacgata agtaaagata tcttcggcga
     4321 atggaacgtg atcagagaca aatggaacgc ggagtatgac gatattcacc tgaagaagaa
     4381 ggctgtcgta acggagaagt acgaggatga tcgcaggaaa agcttcaaaa agatcggaag
     4441 tttcagcctg gaacagttgc aggagtatgc tgacgccgat cttagcgtcg tcgagaagtt
     4501 gaaggagata atcatccaaa aggtcgacga gatatataaa gtctatggat caagtgaaaa
     4561 actgttcgac gccgacttcg ttttggagaa gtccctgaag aagaacgacg ctgttgttgc
     4621 cattatgaag gatctgctcg acagcgtgaa gagtttcgag aactatatta aggctttttt
     4681 cggggagggg aaggagacta acagagatga gtccttctac ggagacttcg tcctcgcgta
     4741 cgatatactc cttaaggtag accacatcta cgacgcaatc agaaattacg tgacacaaaa
     4801 gccgtacagc aaggacaagt tcaaactcta cttccagaac ccccagttca tgggcggctg
     4861 ggacaaggac aaggaaacgg attacagggc tacgatcctg aggtatggtt caaaatacta
     4921 cttggcgatt atggacaaga agtacgccaa gtgtctccag aagattgaca aagacgatgt
     4981 caatggcaat tatgagaaga tcaactacaa gctgcttccg ggtccgaaca agatgctccc
     5041 aaaggttttc ttcagcaaga aatggatggc ctactataac ccaagcgagg acatccagaa
     5101 gatttataag aacggtacgt tcaagaaggg cgacatgttc aatcttaacg actgtcacaa
     5161 gctgatcgac ttcttcaaag actcaattag ccggtaccca aagtggtcta acgcctatga
     5221 cttcaacttt tcggaaaccg agaagtacaa ggatatagcc ggattttata gagaggtgga
     5281 agagcagggc tacaaggtgt cattcgagtc cgccagcaag aaggaagtgg acaagctcgt
     5341 ggaagagggt aagctctaca tgttccagat ttataataaa gactttagcg ataagagcca
     5401 cgggacacct aatctccaca caatgtattt caagctgctc ttcgacgaga ataaccacgg
     5461 ccaaatcagg ttgtcaggag gggctgaact cttcatgcgg cgcgctagcc ttaagaagga
     5521 ggagcttgta gtccaccctg cgaatagtcc aattgcgaat aagaacccgg acaatcctaa
     5581 aaagactaca acattgagct acgacgtgta caaggataag aggttttccg aggatcagta
     5641 cgagctccac atcccgattg cgatcaacaa gtgcccaaag aatattttca agataaacac
     5701 agaggtgcgt gtactcctga agcatgacga caatccttac gtcattggga ttgcccgggg
     5761 cgagaggaac ctcctctata ttgtggtggt ggacgggaag gggaacatag tcgaacagta
     5821 ctcccttaac gaaataatta acaatttcaa cggcatccgt atcaagaccg actaccattc
     5881 gttgctggac aagaaggaga aggagagatt tgaggcgcgg caaaattgga caagtatcga
     5941 gaacatcaag gaactcaaag caggttatat ctctcaagtt gtgcataaga tatgcgagct
     6001 ggttgagaag tatgacgcag tgatcgctct tgaggacctc aactcgggct ttaagaattc
     6061 tagagttaaa gtggagaagc aggtctatca aaagttcgag aagatgctta tagataagct
     6121 caactacatg gtcgataaga aatcgaaccc atgtgccacc ggcggcgcac tcaaaggtta
     6181 ccaaataaca aacaaattcg agtccttcaa atcgatgagt actcagaatg ggttcatatt
     6241 ttatataccg gcgtggctta cgtctaagat cgacccgtca actggttttg tcaacctgtt
     6301 gaagacgaaa tacacgtcca ttgccgattc gaaaaagttc atatctagtt ttgatcgtat
     6361 tatgtacgtc ccagaggaag atcttttcga gtttgctctc gactacaaaa acttttcgcg
     6421 gaccgatgcg gattacatta aaaaatggaa actctattcg tacggcaaca gaatcaggat
     6481 ttttcgcaac cctaagaaga ataacgtctt tgattgggag gaagtttgct tgactagcgc
     6541 gtacaaggag ctctttaata agtatggcat taactaccaa cagggtgata tcagagcact
     6601 gctttgcgaa caatctgaca aggctttcta ctcatccttc atggctttga tgagcctgat
     6661 gctccagatg agaaattcaa ttacaggcag aaccgacgtg gatttcttga tctccccggt
     6721 taaaaattct gatggcatct tttacgatag caggaactat gaagcgcaag agaatgcgat
     6781 tctgccaaaa aatgcagacg ccaacggtgc ctataacatc gccaggaaag tcctgtgggc
     6841 gatcggccag ttcaaaaagg ccgaagacga aaaattggac aaggtcaaaa tcgctatcag
     6901 caacaaagag tggctggagt atgctcagac atccgtaaag cataagcgtc ctgctgccac
     6961 caaaaaggcc ggacaggcta agaaaaagaa gtgagacgac tagtggcggc cgccgacgtc
     7021 cgatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc
     7081 gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg
     7141 catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata
     7201 cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc
     7261 tatgttacta gatcgggaat tgatcccccc tcgacagctt ccggaaaggg cgaattcgca
     7321 actttgtata caaaagttga acgagaaacg taaaatgata taaatatcaa tatattaaat
     7381 tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat ccagtcacta
     7441 tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg
     7501 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
     7561 tcatgcctcc tc
//
