LOCUS       sequence-392890-        7947 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 with 3xFLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7947)
  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 7947)
  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-CBE1
FEATURES             Location/Qualifiers
     source          1..7947
                     /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             3227..3292
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3299..3319
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7025..7072
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7094..7342
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7355..7375
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      7397..7649
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    7694..7817
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(7834..7853)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7835..7853)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7858..7874)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7858..7874)
                     /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 accatggatt acaaggacca
     3241 cgacggggat tacaaggacc acgacattga ttacaaggat gatgatgaca agatggctcc
     3301 gaagaagaag aggaaggttg gcatccacgg ggtgccagct gcttcaaagc tcgagaaatt
     3361 caccaactgt tattcgttga gcaaaacact gcggtttaaa gcgattccag tcggcaagac
     3421 tcaagagaat atagacaata agcggctgtt ggtggaagat gaaaagcgcg cggaagacta
     3481 caaaggggtg aagaagttgt tggacagata ctacctctct tttatcaatg atgtcttgca
     3541 ctcaatcaaa ttgaagaatc tgaacaacta catctccctc ttcagaaaga aaacaaggac
     3601 agaaaaggag aataaggaac ttgaaaattt ggagatcaat ctgaggaaag agatcgcgaa
     3661 agcctttaaa ggcaacgaag gatacaaaag tctgttcaag aaggatataa ttgagacaat
     3721 tttgccagag ttcctcgatg acaaggacga gattgcgctg gtcaattcgt tcaacggatt
     3781 cacaacagca ttcacaggct tctttcgcaa tcgggaaaat atgttctctg aggaggcaaa
     3841 gtccacttct attgcgttca ggtgtatcaa tgagaatctc actaggtaca tttccaacat
     3901 ggatatcttt gagaaggttg acgcaatttt tgacaagcac gaagttcagg agattaagga
     3961 gaagatcctc aattccgatt atgacgttga ggacttcttc gaaggtgagt tttttaattt
     4021 cgtgctcact caagagggta tcgacgtgta taatgcgatc atcggtgggt tcgtgactga
     4081 gtccggtgaa aagattaagg gattgaacga gtatatcaac ctttacaacc aaaagacgaa
     4141 acagaagctg ccaaagttca agcctcttta caaacaggtt ctttcagacc gcgagtcact
     4201 ctcgttctat ggggagggct acacttcgga tgaggaagtc ctggaggtgt tcaggaatac
     4261 tctcaataag aattcggaga ttttctcttc tataaaaaaa ctggaaaagt tgtttaagaa
     4321 ttttgacgaa tactctagcg ccggcatatt tgtgaaaaac ggcccggcca tatcaacgat
     4381 aagtaaagat atcttcggcg aatggaacgt gatcagagac aaatggaacg cggagtatga
     4441 cgatattcac ctgaagaaga aggctgtcgt aacggagaag tacgaggatg atcgcaggaa
     4501 aagcttcaaa aagatcggaa gtttcagcct ggaacagttg caggagtatg ctgacgccga
     4561 tcttagcgtc gtcgagaagt tgaaggagat aatcatccaa aaggtcgacg agatatataa
     4621 agtctatgga tcaagtgaaa aactgttcga cgccgacttc gttttggaga agtccctgaa
     4681 gaagaacgac gctgttgttg ccattatgaa ggatctgctc gacagcgtga agagtttcga
     4741 gaactatatt aaggcttttt tcggggaggg gaaggagact aacagagatg agtccttcta
     4801 cggagacttc gtcctcgcgt acgatatact ccttaaggta gaccacatct acgacgcaat
     4861 cagaaattac gtgacacaaa agccgtacag caaggacaag ttcaaactct acttccagaa
     4921 cccccagttc atgggcggct gggacaagga caaggaaacg gattacaggg ctacgatcct
     4981 gaggtatggt tcaaaatact acttggcgat tatggacaag aagtacgcca agtgtctcca
     5041 gaagattgac aaagacgatg tcaatggcaa ttatgagaag atcaactaca agctgcttcc
     5101 gggtccgaac aagatgctcc caaaggtttt cttcagcaag aaatggatgg cctactataa
     5161 cccaagcgag gacatccaga agatttataa gaacggtacg ttcaagaagg gcgacatgtt
     5221 caatcttaac gactgtcaca agctgatcga cttcttcaaa gactcaatta gccggtaccc
     5281 aaagtggtct aacgcctatg acttcaactt ttcggaaacc gagaagtaca aggatatagc
     5341 cggattttat agagaggtgg aagagcaggg ctacaaggtg tcattcgagt ccgccagcaa
     5401 gaaggaagtg gacaagctcg tggaagaggg taagctctac atgttccaga tttataataa
     5461 agactttagc gataagagcc acgggacacc taatctccac acaatgtatt tcaagctgct
     5521 cttcgacgag aataaccacg gccaaatcag gttgtcagga ggggctgaac tcttcatgcg
     5581 gcgcgctagc cttaagaagg aggagcttgt agtccaccct gcgaatagtc caattgcgaa
     5641 taagaacccg gacaatccta aaaagactac aacattgagc tacgacgtgt acaaggataa
     5701 gaggttttcc gaggatcagt acgagctcca catcccgatt gcgatcaaca agtgcccaaa
     5761 gaatattttc aagataaaca cagaggtgcg tgtactcctg aagcatgacg acaatcctta
     5821 cgtcattggg attgcccggg gcgagaggaa cctcctctat attgtggtgg tggacgggaa
     5881 ggggaacata gtcgaacagt actcccttaa cgaaataatt aacaatttca acggcatccg
     5941 tatcaagacc gactaccatt cgttgctgga caagaaggag aaggagagat ttgaggcgcg
     6001 gcaaaattgg acaagtatcg agaacatcaa ggaactcaaa gcaggttata tctctcaagt
     6061 tgtgcataag atatgcgagc tggttgagaa gtatgacgca gtgatcgctc ttgaggacct
     6121 caactcgggc tttaagaatt ctagagttaa agtggagaag caggtctatc aaaagttcga
     6181 gaagatgctt atagataagc tcaactacat ggtcgataag aaatcgaacc catgtgccac
     6241 cggcggcgca ctcaaaggtt accaaataac aaacaaattc gagtccttca aatcgatgag
     6301 tactcagaat gggttcatat tttatatacc ggcgtggctt acgtctaaga tcgacccgtc
     6361 aactggtttt gtcaacctgt tgaagacgaa atacacgtcc attgccgatt cgaaaaagtt
     6421 catatctagt tttgatcgta ttatgtacgt cccagaggaa gatcttttcg agtttgctct
     6481 cgactacaaa aacttttcgc ggaccgatgc ggattacatt aaaaaatgga aactctattc
     6541 gtacggcaac agaatcagga tttttcgcaa ccctaagaag aataacgtct ttgattggga
     6601 ggaagtttgc ttgactagcg cgtacaagga gctctttaat aagtatggca ttaactacca
     6661 acagggtgat atcagagcac tgctttgcga acaatctgac aaggctttct actcatcctt
     6721 catggctttg atgagcctga tgctccagat gagaaattca attacaggca gaaccgacgt
     6781 ggatttcttg atctccccgg ttaaaaattc tgatggcatc ttttacgata gcaggaacta
     6841 tgaagcgcaa gagaatgcga ttctgccaaa aaatgcagac gccaacggtg cctataacat
     6901 cgccaggaaa gtcctgtggg cgatcggcca gttcaaaaag gccgaagacg aaaaattgga
     6961 caaggtcaaa atcgctatca gcaacaaaga gtggctggag tatgctcaga catccgtaaa
     7021 gcataagcgg ccagcggcga cgaagaaggc ggggcaggcg aagaagaaga agggagacgg
     7081 atcaggcggg tcaacaaacc tcagtgacat catagagaag gaaactggta agcaactggt
     7141 tattcaagag agcattctga tgctccctga ggaagtcgag gaagttatag gaaacaagcc
     7201 tgagagcgat atactcgtgc atacggcgta tgacgagagc acggatgaaa atgtgatgct
     7261 cttgaccagc gatgcgccag aatataaacc ctgggcactg gttattcagg actctaacgg
     7321 ggaaaataaa atcaagatgc tctcaggtgg ctccccaaag aagaaacgca aggtttagtg
     7381 gcggccgccg acgtccgatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc
     7441 tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta agcatgtaat
     7501 aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca
     7561 attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc
     7621 gcgcgcggtg tcatctatgt tactagatcg ggaattgatc ccccctcgac agcttccgga
     7681 aagggcgaat tcgcaacttt gtatacaaaa gttgaacgag aaacgtaaaa tgatataaat
     7741 atcaatatat taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa
     7801 catatccagt cactatgcca tccagctgat atcccctata gtgagtcgta ttacatggtc
     7861 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     7921 gataaaaata tatcatcatg cctcctc
//
