LOCUS       sequence-392913-        7878 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dLbCas12a-D156R based cytosine base editor, hA3A/Y130F fused to the 
            N terminal of dLbCas12a-D156R with the linker of XTEN.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7878)
  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 7878)
  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-CBE2
FEATURES             Location/Qualifiers
     source          1..7878
                     /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             3230..3250
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6956..7003
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7025..7273
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7286..7306
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      7328..7580
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    7625..7748
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(7765..7784)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7766..7784)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7789..7805)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7789..7805)
                     /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 ccgaattcgc ccttcatgga
     2581 ggcctctccg gcttcaggtc ctcgccatct catggacccc catatcttta cctccaattt
     2641 caataacggc attggcaggc acaagacata cctctgttat gaggtcgagc ggctcgataa
     2701 cggaacctca gtgaagatgg accaacacag gggtttcctt cataaccaag cgaaaaatct
     2761 cctttgcggt ttctatgggc ggcacgcgga gctgagattc ttggatttgg tcccgagcct
     2821 ccagctggat cccgcgcaaa tataccgcgt cacttggttc atatcatgga gcccgtgctt
     2881 tagctggggg tgtgccggtg aggttcgcgc gttcctccag gaaaacaccc atgttcgcct
     2941 ccggattttc gcagcgagga ttttcgacta tgatccactt tacaaagagg ccctccaaat
     3001 gcttagagat gctggggctc aagtttcaat tatgacatac gacgagttta agcattgttg
     3061 ggacactttt gtcgatcatc agggttgtcc cttccagccg tgggatggtc ttgatgagca
     3121 ttcgcaagct ttgagtggca ggcttcgcgc gatacttcag aaccagggta actcaggttc
     3181 tgagacccct ggcacaagtg agtcagcaac acccgagtcc accatggctc cgaagaagaa
     3241 gaggaaggtt ggcatccacg gggtgccagc tgcttcaaag ctcgagaaat tcaccaactg
     3301 ttattcgttg agcaaaacac tgcggtttaa agcgattcca gtcggcaaga ctcaagagaa
     3361 tatagacaat aagcggctgt tggtggaaga tgaaaagcgc gcggaagact acaaaggggt
     3421 gaagaagttg ttggacagat actacctctc ttttatcaat gatgtcttgc actcaatcaa
     3481 attgaagaat ctgaacaact acatctccct cttcagaaag aaaacaagga cagaaaagga
     3541 gaataaggaa cttgaaaatt tggagatcaa tctgaggaaa gagatcgcga aagcctttaa
     3601 aggcaacgaa ggatacaaaa gtctgttcaa gaaggatata attgagacaa ttttgccaga
     3661 gttcctcgat gacaaggacg agattgcgct ggtcaattcg ttcaacggat tcacaacagc
     3721 attcacaggc ttctttcgca atcgggaaaa tatgttctct gaggaggcaa agtccacttc
     3781 tattgcgttc aggtgtatca atgagaatct cactaggtac atttccaaca tggatatctt
     3841 tgagaaggtt gacgcaattt ttgacaagca cgaagttcag gagattaagg agaagatcct
     3901 caattccgat tatgacgttg aggacttctt cgaaggtgag ttttttaatt tcgtgctcac
     3961 tcaagagggt atcgacgtgt ataatgcgat catcggtggg ttcgtgactg agtccggtga
     4021 aaagattaag ggattgaacg agtatatcaa cctttacaac caaaagacga aacagaagct
     4081 gccaaagttc aagcctcttt acaaacaggt tctttcagac cgcgagtcac tctcgttcta
     4141 tggggagggc tacacttcgg atgaggaagt cctggaggtg ttcaggaata ctctcaataa
     4201 gaattcggag attttctctt ctataaaaaa actggaaaag ttgtttaaga attttgacga
     4261 atactctagc gccggcatat ttgtgaaaaa cggcccggcc atatcaacga taagtaaaga
     4321 tatcttcggc gaatggaacg tgatcagaga caaatggaac gcggagtatg acgatattca
     4381 cctgaagaag aaggctgtcg taacggagaa gtacgaggat gatcgcagga aaagcttcaa
     4441 aaagatcgga agtttcagcc tggaacagtt gcaggagtat gctgacgccg atcttagcgt
     4501 cgtcgagaag ttgaaggaga taatcatcca aaaggtcgac gagatatata aagtctatgg
     4561 atcaagtgaa aaactgttcg acgccgactt cgttttggag aagtccctga agaagaacga
     4621 cgctgttgtt gccattatga aggatctgct cgacagcgtg aagagtttcg agaactatat
     4681 taaggctttt ttcggggagg ggaaggagac taacagagat gagtccttct acggagactt
     4741 cgtcctcgcg tacgatatac tccttaaggt agaccacatc tacgacgcaa tcagaaatta
     4801 cgtgacacaa aagccgtaca gcaaggacaa gttcaaactc tacttccaga acccccagtt
     4861 catgggcggc tgggacaagg acaaggaaac ggattacagg gctacgatcc tgaggtatgg
     4921 ttcaaaatac tacttggcga ttatggacaa gaagtacgcc aagtgtctcc agaagattga
     4981 caaagacgat gtcaatggca attatgagaa gatcaactac aagctgcttc cgggtccgaa
     5041 caagatgctc ccaaaggttt tcttcagcaa gaaatggatg gcctactata acccaagcga
     5101 ggacatccag aagatttata agaacggtac gttcaagaag ggcgacatgt tcaatcttaa
     5161 cgactgtcac aagctgatcg acttcttcaa agactcaatt agccggtacc caaagtggtc
     5221 taacgcctat gacttcaact tttcggaaac cgagaagtac aaggatatag ccggatttta
     5281 tagagaggtg gaagagcagg gctacaaggt gtcattcgag tccgccagca agaaggaagt
     5341 ggacaagctc gtggaagagg gtaagctcta catgttccag atttataata aagactttag
     5401 cgataagagc cacgggacac ctaatctcca cacaatgtat ttcaagctgc tcttcgacga
     5461 gaataaccac ggccaaatca ggttgtcagg aggggctgaa ctcttcatgc ggcgcgctag
     5521 ccttaagaag gaggagcttg tagtccaccc tgcgaatagt ccaattgcga ataagaaccc
     5581 ggacaatcct aaaaagacta caacattgag ctacgacgtg tacaaggata agaggttttc
     5641 cgaggatcag tacgagctcc acatcccgat tgcgatcaac aagtgcccaa agaatatttt
     5701 caagataaac acagaggtgc gtgtactcct gaagcatgac gacaatcctt acgtcattgg
     5761 gattgcccgg ggcgagagga acctcctcta tattgtggtg gtggacggga aggggaacat
     5821 agtcgaacag tactccctta acgaaataat taacaatttc aacggcatcc gtatcaagac
     5881 cgactaccat tcgttgctgg acaagaagga gaaggagaga tttgaggcgc ggcaaaattg
     5941 gacaagtatc gagaacatca aggaactcaa agcaggttat atctctcaag ttgtgcataa
     6001 gatatgcgag ctggttgaga agtatgacgc agtgatcgct cttgaggacc tcaactcggg
     6061 ctttaagaat tctagagtta aagtggagaa gcaggtctat caaaagttcg agaagatgct
     6121 tatagataag ctcaactaca tggtcgataa gaaatcgaac ccatgtgcca ccggcggcgc
     6181 actcaaaggt taccaaataa caaacaaatt cgagtccttc aaatcgatga gtactcagaa
     6241 tgggttcata ttttatatac cggcgtggct tacgtctaag atcgacccgt caactggttt
     6301 tgtcaacctg ttgaagacga aatacacgtc cattgccgat tcgaaaaagt tcatatctag
     6361 ttttgatcgt attatgtacg tcccagagga agatcttttc gagtttgctc tcgactacaa
     6421 aaacttttcg cggaccgatg cggattacat taaaaaatgg aaactctatt cgtacggcaa
     6481 cagaatcagg atttttcgca accctaagaa gaataacgtc tttgattggg aggaagtttg
     6541 cttgactagc gcgtacaagg agctctttaa taagtatggc attaactacc aacagggtga
     6601 tatcagagca ctgctttgcg aacaatctga caaggctttc tactcatcct tcatggcttt
     6661 gatgagcctg atgctccaga tgagaaattc aattacaggc agaaccgacg tggatttctt
     6721 gatctccccg gttaaaaatt ctgatggcat cttttacgat agcaggaact atgaagcgca
     6781 agagaatgcg attctgccaa aaaatgcaga cgccaacggt gcctataaca tcgccaggaa
     6841 agtcctgtgg gcgatcggcc agttcaaaaa ggccgaagac gaaaaattgg acaaggtcaa
     6901 aatcgctatc agcaacaaag agtggctgga gtatgctcag acatccgtaa agcataagcg
     6961 gccagcggcg acgaagaagg cggggcaggc gaagaagaag aagggagacg gatcaggcgg
     7021 gtcaacaaac ctcagtgaca tcatagagaa ggaaactggt aagcaactgg ttattcaaga
     7081 gagcattctg atgctccctg aggaagtcga ggaagttata ggaaacaagc ctgagagcga
     7141 tatactcgtg catacggcgt atgacgagag cacggatgaa aatgtgatgc tcttgaccag
     7201 cgatgcgcca gaatataaac cctgggcact ggttattcag gactctaacg gggaaaataa
     7261 aatcaagatg ctctcaggtg gctccccaaa gaagaaacgc aaggtttagt ggcggccgcc
     7321 gacgtccgat cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg
     7381 tcttgcgatg attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat
     7441 gtaatgcatg acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat
     7501 ttaatacgcg atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt
     7561 gtcatctatg ttactagatc gggaattgat cccccctcga cagcttccgg aaagggcgaa
     7621 ttcgcaactt tgtatacaaa agttgaacga gaaacgtaaa atgatataaa tatcaatata
     7681 ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag
     7741 tcactatgcc atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt
     7801 tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     7861 atatcatcat gcctcctc
//
