LOCUS       sequence-392981-        7920 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 (GGGGS)x6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7920)
  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 7920)
  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-CBE4
FEATURES             Location/Qualifiers
     source          1..7920
                     /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             3272..3292
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6998..7045
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7067..7315
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7328..7348
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      7370..7622
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    7667..7790
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(7807..7826)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7808..7826)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7831..7847)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7831..7847)
                     /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 acgggggcgg
     3181 gggctcaggc gggggcggga gcggcggcgg gggctctggg ggcggcggca gcggcggggg
     3241 cggcagcggg ggcggcgggt cgaccatggc tcctaagaag aagcggaagg ttggtattca
     3301 cggggtgcct gcggcttcaa agctcgagaa attcaccaac tgttattcgt tgagcaaaac
     3361 actgcggttt aaagcgattc cagtcggcaa gactcaagag aatatagaca ataagcggct
     3421 gttggtggaa gatgaaaagc gcgcggaaga ctacaaaggg gtgaagaagt tgttggacag
     3481 atactacctc tcttttatca atgatgtctt gcactcaatc aaattgaaga atctgaacaa
     3541 ctacatctcc ctcttcagaa agaaaacaag gacagaaaag gagaataagg aacttgaaaa
     3601 tttggagatc aatctgagga aagagatcgc gaaagccttt aaaggcaacg aaggatacaa
     3661 aagtctgttc aagaaggata taattgagac aattttgcca gagttcctcg atgacaagga
     3721 cgagattgcg ctggtcaatt cgttcaacgg attcacaaca gcattcacag gcttctttcg
     3781 caatcgggaa aatatgttct ctgaggaggc aaagtccact tctattgcgt tcaggtgtat
     3841 caatgagaat ctcactaggt acatttccaa catggatatc tttgagaagg ttgacgcaat
     3901 ttttgacaag cacgaagttc aggagattaa ggagaagatc ctcaattccg attatgacgt
     3961 tgaggacttc ttcgaaggtg agttttttaa tttcgtgctc actcaagagg gtatcgacgt
     4021 gtataatgcg atcatcggtg ggttcgtgac tgagtccggt gaaaagatta agggattgaa
     4081 cgagtatatc aacctttaca accaaaagac gaaacagaag ctgccaaagt tcaagcctct
     4141 ttacaaacag gttctttcag accgcgagtc actctcgttc tatggggagg gctacacttc
     4201 ggatgaggaa gtcctggagg tgttcaggaa tactctcaat aagaattcgg agattttctc
     4261 ttctataaaa aaactggaaa agttgtttaa gaattttgac gaatactcta gcgccggcat
     4321 atttgtgaaa aacggcccgg ccatatcaac gataagtaaa gatatcttcg gcgaatggaa
     4381 cgtgatcaga gacaaatgga acgcggagta tgacgatatt cacctgaaga agaaggctgt
     4441 cgtaacggag aagtacgagg atgatcgcag gaaaagcttc aaaaagatcg gaagtttcag
     4501 cctggaacag ttgcaggagt atgctgacgc cgatcttagc gtcgtcgaga agttgaagga
     4561 gataatcatc caaaaggtcg acgagatata taaagtctat ggatcaagtg aaaaactgtt
     4621 cgacgccgac ttcgttttgg agaagtccct gaagaagaac gacgctgttg ttgccattat
     4681 gaaggatctg ctcgacagcg tgaagagttt cgagaactat attaaggctt ttttcgggga
     4741 ggggaaggag actaacagag atgagtcctt ctacggagac ttcgtcctcg cgtacgatat
     4801 actccttaag gtagaccaca tctacgacgc aatcagaaat tacgtgacac aaaagccgta
     4861 cagcaaggac aagttcaaac tctacttcca gaacccccag ttcatgggcg gctgggacaa
     4921 ggacaaggaa acggattaca gggctacgat cctgaggtat ggttcaaaat actacttggc
     4981 gattatggac aagaagtacg ccaagtgtct ccagaagatt gacaaagacg atgtcaatgg
     5041 caattatgag aagatcaact acaagctgct tccgggtccg aacaagatgc tcccaaaggt
     5101 tttcttcagc aagaaatgga tggcctacta taacccaagc gaggacatcc agaagattta
     5161 taagaacggt acgttcaaga agggcgacat gttcaatctt aacgactgtc acaagctgat
     5221 cgacttcttc aaagactcaa ttagccggta cccaaagtgg tctaacgcct atgacttcaa
     5281 cttttcggaa accgagaagt acaaggatat agccggattt tatagagagg tggaagagca
     5341 gggctacaag gtgtcattcg agtccgccag caagaaggaa gtggacaagc tcgtggaaga
     5401 gggtaagctc tacatgttcc agatttataa taaagacttt agcgataaga gccacgggac
     5461 acctaatctc cacacaatgt atttcaagct gctcttcgac gagaataacc acggccaaat
     5521 caggttgtca ggaggggctg aactcttcat gcggcgcgct agccttaaga aggaggagct
     5581 tgtagtccac cctgcgaata gtccaattgc gaataagaac ccggacaatc ctaaaaagac
     5641 tacaacattg agctacgacg tgtacaagga taagaggttt tccgaggatc agtacgagct
     5701 ccacatcccg attgcgatca acaagtgccc aaagaatatt ttcaagataa acacagaggt
     5761 gcgtgtactc ctgaagcatg acgacaatcc ttacgtcatt gggattgccc ggggcgagag
     5821 gaacctcctc tatattgtgg tggtggacgg gaaggggaac atagtcgaac agtactccct
     5881 taacgaaata attaacaatt tcaacggcat ccgtatcaag accgactacc attcgttgct
     5941 ggacaagaag gagaaggaga gatttgaggc gcggcaaaat tggacaagta tcgagaacat
     6001 caaggaactc aaagcaggtt atatctctca agttgtgcat aagatatgcg agctggttga
     6061 gaagtatgac gcagtgatcg ctcttgagga cctcaactcg ggctttaaga attctagagt
     6121 taaagtggag aagcaggtct atcaaaagtt cgagaagatg cttatagata agctcaacta
     6181 catggtcgat aagaaatcga acccatgtgc caccggcggc gcactcaaag gttaccaaat
     6241 aacaaacaaa ttcgagtcct tcaaatcgat gagtactcag aatgggttca tattttatat
     6301 accggcgtgg cttacgtcta agatcgaccc gtcaactggt tttgtcaacc tgttgaagac
     6361 gaaatacacg tccattgccg attcgaaaaa gttcatatct agttttgatc gtattatgta
     6421 cgtcccagag gaagatcttt tcgagtttgc tctcgactac aaaaactttt cgcggaccga
     6481 tgcggattac attaaaaaat ggaaactcta ttcgtacggc aacagaatca ggatttttcg
     6541 caaccctaag aagaataacg tctttgattg ggaggaagtt tgcttgacta gcgcgtacaa
     6601 ggagctcttt aataagtatg gcattaacta ccaacagggt gatatcagag cactgctttg
     6661 cgaacaatct gacaaggctt tctactcatc cttcatggct ttgatgagcc tgatgctcca
     6721 gatgagaaat tcaattacag gcagaaccga cgtggatttc ttgatctccc cggttaaaaa
     6781 ttctgatggc atcttttacg atagcaggaa ctatgaagcg caagagaatg cgattctgcc
     6841 aaaaaatgca gacgccaacg gtgcctataa catcgccagg aaagtcctgt gggcgatcgg
     6901 ccagttcaaa aaggccgaag acgaaaaatt ggacaaggtc aaaatcgcta tcagcaacaa
     6961 agagtggctg gagtatgctc agacatccgt aaagcataag cggccagcgg cgacgaagaa
     7021 ggcggggcag gcgaagaaga agaagggaga cggatcaggc gggtcaacaa acctcagtga
     7081 catcatagag aaggaaactg gtaagcaact ggttattcaa gagagcattc tgatgctccc
     7141 tgaggaagtc gaggaagtta taggaaacaa gcctgagagc gatatactcg tgcatacggc
     7201 gtatgacgag agcacggatg aaaatgtgat gctcttgacc agcgatgcgc cagaatataa
     7261 accctgggca ctggttattc aggactctaa cggggaaaat aaaatcaaga tgctctcagg
     7321 tggctcccca aagaagaaac gcaaggttta gtggcggccg ccgacgtccg atcgttcaaa
     7381 catttggcaa taaagtttct taagattgaa tcctgttgcc ggtcttgcga tgattatcat
     7441 ataatttctg ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt
     7501 tatgagatgg gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa
     7561 caaaatatag cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactaga
     7621 tcgggaattg atcccccctc gacagcttcc ggaaagggcg aattcgcaac tttgtataca
     7681 aaagttgaac gagaaacgta aaatgatata aatatcaata tattaaatta gattttgcat
     7741 aaaaaacaga ctacataata ctgtaaaaca caacatatcc agtcactatg ccatccagct
     7801 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     7861 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgcctcctc
//
