LOCUS       sequence-392986-        7602 bp    DNA     circular SYN 14-APR-2025
DEFINITION  dLbCas12a-D156R based adenine base editor, ecTadA8e fused to the N 
            terminal of dLbCas12a-D156R with the linker of (GGGGS)x8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7602)
  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 7602)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-ABE5
FEATURES             Location/Qualifiers
     source          1..7602
                     /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             3248..3268
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6974..7021
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      7052..7304
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    7349..7472
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(7489..7508)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7490..7508)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7513..7529)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7513..7529)
                     /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 agggggaggg tcgggaggtg gtggcagcgg aggaggcggt agcggaggtg gaggatcagg
     3181 tggcggaggg tcgggagggg gcggttctgg tggtgggggc tcgggtggag gagggagcac
     3241 catggctcct aagaagaagc ggaaggttgg tattcacggg gtgcctgcgg cttcaaagct
     3301 cgagaaattc accaactgtt attcgttgag caaaacactg cggtttaaag cgattccagt
     3361 cggcaagact caagagaata tagacaataa gcggctgttg gtggaagatg aaaagcgcgc
     3421 ggaagactac aaaggggtga agaagttgtt ggacagatac tacctctctt ttatcaatga
     3481 tgtcttgcac tcaatcaaat tgaagaatct gaacaactac atctccctct tcagaaagaa
     3541 aacaaggaca gaaaaggaga ataaggaact tgaaaatttg gagatcaatc tgaggaaaga
     3601 gatcgcgaaa gcctttaaag gcaacgaagg atacaaaagt ctgttcaaga aggatataat
     3661 tgagacaatt ttgccagagt tcctcgatga caaggacgag attgcgctgg tcaattcgtt
     3721 caacggattc acaacagcat tcacaggctt ctttcgcaat cgggaaaata tgttctctga
     3781 ggaggcaaag tccacttcta ttgcgttcag gtgtatcaat gagaatctca ctaggtacat
     3841 ttccaacatg gatatctttg agaaggttga cgcaattttt gacaagcacg aagttcagga
     3901 gattaaggag aagatcctca attccgatta tgacgttgag gacttcttcg aaggtgagtt
     3961 ttttaatttc gtgctcactc aagagggtat cgacgtgtat aatgcgatca tcggtgggtt
     4021 cgtgactgag tccggtgaaa agattaaggg attgaacgag tatatcaacc tttacaacca
     4081 aaagacgaaa cagaagctgc caaagttcaa gcctctttac aaacaggttc tttcagaccg
     4141 cgagtcactc tcgttctatg gggagggcta cacttcggat gaggaagtcc tggaggtgtt
     4201 caggaatact ctcaataaga attcggagat tttctcttct ataaaaaaac tggaaaagtt
     4261 gtttaagaat tttgacgaat actctagcgc cggcatattt gtgaaaaacg gcccggccat
     4321 atcaacgata agtaaagata tcttcggcga atggaacgtg atcagagaca aatggaacgc
     4381 ggagtatgac gatattcacc tgaagaagaa ggctgtcgta acggagaagt acgaggatga
     4441 tcgcaggaaa agcttcaaaa agatcggaag tttcagcctg gaacagttgc aggagtatgc
     4501 tgacgccgat cttagcgtcg tcgagaagtt gaaggagata atcatccaaa aggtcgacga
     4561 gatatataaa gtctatggat caagtgaaaa actgttcgac gccgacttcg ttttggagaa
     4621 gtccctgaag aagaacgacg ctgttgttgc cattatgaag gatctgctcg acagcgtgaa
     4681 gagtttcgag aactatatta aggctttttt cggggagggg aaggagacta acagagatga
     4741 gtccttctac ggagacttcg tcctcgcgta cgatatactc cttaaggtag accacatcta
     4801 cgacgcaatc agaaattacg tgacacaaaa gccgtacagc aaggacaagt tcaaactcta
     4861 cttccagaac ccccagttca tgggcggctg ggacaaggac aaggaaacgg attacagggc
     4921 tacgatcctg aggtatggtt caaaatacta cttggcgatt atggacaaga agtacgccaa
     4981 gtgtctccag aagattgaca aagacgatgt caatggcaat tatgagaaga tcaactacaa
     5041 gctgcttccg ggtccgaaca agatgctccc aaaggttttc ttcagcaaga aatggatggc
     5101 ctactataac ccaagcgagg acatccagaa gatttataag aacggtacgt tcaagaaggg
     5161 cgacatgttc aatcttaacg actgtcacaa gctgatcgac ttcttcaaag actcaattag
     5221 ccggtaccca aagtggtcta acgcctatga cttcaacttt tcggaaaccg agaagtacaa
     5281 ggatatagcc ggattttata gagaggtgga agagcagggc tacaaggtgt cattcgagtc
     5341 cgccagcaag aaggaagtgg acaagctcgt ggaagagggt aagctctaca tgttccagat
     5401 ttataataaa gactttagcg ataagagcca cgggacacct aatctccaca caatgtattt
     5461 caagctgctc ttcgacgaga ataaccacgg ccaaatcagg ttgtcaggag gggctgaact
     5521 cttcatgcgg cgcgctagcc ttaagaagga ggagcttgta gtccaccctg cgaatagtcc
     5581 aattgcgaat aagaacccgg acaatcctaa aaagactaca acattgagct acgacgtgta
     5641 caaggataag aggttttccg aggatcagta cgagctccac atcccgattg cgatcaacaa
     5701 gtgcccaaag aatattttca agataaacac agaggtgcgt gtactcctga agcatgacga
     5761 caatccttac gtcattggga ttgcccgggg cgagaggaac ctcctctata ttgtggtggt
     5821 ggacgggaag gggaacatag tcgaacagta ctcccttaac gaaataatta acaatttcaa
     5881 cggcatccgt atcaagaccg actaccattc gttgctggac aagaaggaga aggagagatt
     5941 tgaggcgcgg caaaattgga caagtatcga gaacatcaag gaactcaaag caggttatat
     6001 ctctcaagtt gtgcataaga tatgcgagct ggttgagaag tatgacgcag tgatcgctct
     6061 tgaggacctc aactcgggct ttaagaattc tagagttaaa gtggagaagc aggtctatca
     6121 aaagttcgag aagatgctta tagataagct caactacatg gtcgataaga aatcgaaccc
     6181 atgtgccacc ggcggcgcac tcaaaggtta ccaaataaca aacaaattcg agtccttcaa
     6241 atcgatgagt actcagaatg ggttcatatt ttatataccg gcgtggctta cgtctaagat
     6301 cgacccgtca actggttttg tcaacctgtt gaagacgaaa tacacgtcca ttgccgattc
     6361 gaaaaagttc atatctagtt ttgatcgtat tatgtacgtc ccagaggaag atcttttcga
     6421 gtttgctctc gactacaaaa acttttcgcg gaccgatgcg gattacatta aaaaatggaa
     6481 actctattcg tacggcaaca gaatcaggat ttttcgcaac cctaagaaga ataacgtctt
     6541 tgattgggag gaagtttgct tgactagcgc gtacaaggag ctctttaata agtatggcat
     6601 taactaccaa cagggtgata tcagagcact gctttgcgaa caatctgaca aggctttcta
     6661 ctcatccttc atggctttga tgagcctgat gctccagatg agaaattcaa ttacaggcag
     6721 aaccgacgtg gatttcttga tctccccggt taaaaattct gatggcatct tttacgatag
     6781 caggaactat gaagcgcaag agaatgcgat tctgccaaaa aatgcagacg ccaacggtgc
     6841 ctataacatc gccaggaaag tcctgtgggc gatcggccag ttcaaaaagg ccgaagacga
     6901 aaaattggac aaggtcaaaa tcgctatcag caacaaagag tggctggagt atgctcagac
     6961 atccgtaaag cataagcgtc ctgctgccac caaaaaggcc ggacaggcta agaaaaagaa
     7021 gtgagacgac tagtggcggc cgccgacgtc cgatcgttca aacatttggc aataaagttt
     7081 cttaagattg aatcctgttg ccggtcttgc gatgattatc atataatttc tgttgaatta
     7141 cgttaagcat gtaataatta acatgtaatg catgacgtta tttatgagat gggtttttat
     7201 gattagagtc ccgcaattat acatttaata cgcgatagaa aacaaaatat agcgcgcaaa
     7261 ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta gatcgggaat tgatcccccc
     7321 tcgacagctt ccggaaaggg cgaattcgca actttgtata caaaagttga acgagaaacg
     7381 taaaatgata taaatatcaa tatattaaat tagattttgc ataaaaaaca gactacataa
     7441 tactgtaaaa cacaacatat ccagtcacta tgccatccag ctgatatccc ctatagtgag
     7501 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     7561 tgttacattg cacaagataa aaatatatca tcatgcctcc tc
//
