LOCUS       sequence-232090-       10080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Conversion of C to T in wheat plants or monocotyledons protoplasts .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10080)
  AUTHORS   Zong Y, Song Q, Li C, Jin S, Zhang D, Wang Y, Qiu JL, Gao C
  TITLE     Efficient C-to-T base editing in plants using a fusion of nCas9 and 
            human APOBEC3A.
  JOURNAL   Nat Biotechnol. 2018 Oct 1. pii: nbt.4261. doi: 10.1038/nbt.4261.
  PUBMED    30272679
REFERENCE   2  (bases 1 to 10080)
  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: "Nat 
            Biotechnol. 2018 Oct 1. pii: nbt.4261. doi: 10.1038/nbt.4261."
COMMENT     Sequence Label: A3A-PBE
FEATURES             Location/Qualifiers
     source          1..10080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             6770..6817
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6839..7087
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7100..7120
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    7194..7370
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(7924..7940)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7924..7940)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7937..7959)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7948..7964
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(7972..7978,7979..7996,7997..8002))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8017..8038
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8155..8172)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8326..8914)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8406..8425)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9085..9876,9877..9945))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     9708..9727
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9946..10050)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gagctcggta cctgacccgg tcgtgcccct ctctagagat aatgagcatt gcatgtctaa
       61 gttataaaaa attaccacat attttttttg tcacacttgt ttgaagtgca gtttatctat
      121 ctttatacat atatttaaac tttactctac gaataatata atctatagta ctacaataat
      181 atcagtgttt tagagaatca tataaatgaa cagttagaca tggtctaaag gacaattgag
      241 tattttgaca acaggactct acagttttat ctttttagtg tgcatgtgtt ctcctttttt
      301 tttgcaaata gcttcaccta tataatactt catccatttt attagtacat ccatttaggg
      361 tttagggtta atggttttta tagactaatt tttttagtac atctatttta ttctatttta
      421 gcctctaaat taagaaaact aaaactctat tttagttttt ttatttaata atttagatat
      481 aaaatagaat aaaataaagt gactaaaaat taaacaaata ccctttaaga aattaaaaaa
      541 actaaggaaa catttttctt gtttcgagta gataatgcca gcctgttaaa cgccgtcgac
      601 gagtctaacg gacaccaacc agcgaaccag cagcgtcgcg tcgggccaag cgaagcagac
      661 ggcacggcat ctctgtcgct gcctctggac ccctctcgat cgagagttcc gctccaccgt
      721 tggacttgct ccgctgtcgg catccagaaa ttgcgtggcg gagcggcaga cgtgagccgg
      781 cacggcaggc ggcctcctcc tcctctcacg gcaccggcag ctacggggga ttcctttccc
      841 accgctcctt cgctttccct tcctcgcccg ccgtaataaa tagacacccc ctccacaccc
      901 tctttcccca acctcgtgtt gttcggagcg cacacacaca caaccagatc tcccccaaat
      961 ccacccgtcg gcacctccgc ttcaaggtac gccgctcgtc ctcccccccc ccccctctct
     1021 accttctcta gatcggcgtt ccggtccatg gttagggccc ggtagttcta cttctgttca
     1081 tgtttgtgtt agatccgtgt ttgtgttaga tccgtgctgc tagcgttcgt acacggatgc
     1141 gacctgtacg tcagacacgt tctgattgct aacttgccag tgtttctctt tggggaatcc
     1201 tgggatggct ctagccgttc cgcagacggg atcgatttca tgattttttt tgtttcgttg
     1261 catagggttt ggtttgccct tttcctttat ttcaatatat gccgtgcact tgtttgtcgg
     1321 gtcatctttt catgcttttt tttgtcttgg ttgtgatgat gtggtctggt tgggcggtcg
     1381 ttctagatcg gagtagaatt aattctgttt caaactacct ggtggattta ttaattttgg
     1441 atctgtatgt gtgtgccata catattcata gttacgaatt gaagatgatg gatggaaata
     1501 tcgatctagg ataggtatac atgttgatgc gggttttact gatgcatata cagagatgct
     1561 ttttgttcgc ttggttgtga tgatgtggtg tggttgggcg gtcgttcatt cgttctagat
     1621 cggagtagaa tactgtttca aactacctgg tgtatttatt aattttggaa ctgtatgtgt
     1681 gtgtcataca tcttcatagt tacgagttta agatggatgg aaatatcgat ctaggatagg
     1741 tatacatgtt gatgtgggtt ttactgatgc atatacatga tggcatatgc agcatctatt
     1801 catatgctct aaccttgagt acctatctat tataataaac aagtatgttt tataattatt
     1861 ttgatcttga tatacttgga tgatggcata tgcagcagct atatgtggat ttttttagcc
     1921 ctgccttcat acgctattta tttgcttggt actgtttctt ttgtcgatgc tcaccctgtt
     1981 gtttggtgtt acttctgcaa agcttgtcga cggatccatg gaggccagcc cggctagcgg
     2041 cccaaggcat ctcatggacc cgcacatctt caccagcaac ttcaacaacg gcatcggcag
     2101 gcacaagacc tacttgtgct acgaggtgga gaggctcgac aacggaacct ccgtgaagat
     2161 ggaccaacac agggggttcc tccacaacca agccaagaac ctcctctgcg gcttctacgg
     2221 caggcacgcc gagttgaggt tcctcgactt ggtgccatcc ctccaactcg atccagccca
     2281 aatctaccgc gtgacctggt tcatctcctg gtccccatgc ttctcctggg gttgcgccgg
     2341 cgaggttcgg gctttcctcc aagaaaacac ccacgtccgc ctccgcattt tcgccgccag
     2401 gatctatgat tacgaccctc tctacaagga ggccctccag atgctgcggg acgccggtgc
     2461 tcaggtgagt atcatgacct acgacgagtt caagcactgc tgggacacct tcgttgacca
     2521 ccagggctgc ccattccaac catgggacgg tctggatgaa cacagccaag ccttgtccgg
     2581 caggctccgg gccatcctcc aaaaccaggg gaactccggg agcgagacgc caggcacctc
     2641 cgagtcggcc accccagaat ctcttaagga caagaagtac tcgatcggcc tcgccatcgg
     2701 gacgaactca gttggctggg ccgtgatcac cgacgagtac aaggtgccct ctaagaagtt
     2761 caaggtcctg gggaacaccg accgccattc catcaagaag aacctcatcg gcgctctcct
     2821 gttcgacagc ggggagaccg ctgaggctac gaggctcaag agaaccgcta ggcgccggta
     2881 cacgagaagg aagaacagga tctgctacct ccaagagatt ttctccaacg agatggccaa
     2941 ggttgacgat tcattcttcc accgcctgga ggagtctttc ctcgtggagg aggataagaa
     3001 gcacgagcgg catcccatct tcggcaacat cgtggacgag gttgcctacc acgagaagta
     3061 ccctacgatc taccatctgc ggaagaagct cgtggactcc accgataagg cggacctcag
     3121 actgatctac ctcgctctgg cccacatgat caagttccgc ggccatttcc tgatcgaggg
     3181 ggatctcaac ccagacaaca gcgatgttga caagctgttc atccaactcg tgcagaccta
     3241 caaccaactc ttcgaggaga acccgatcaa cgcctctggc gtggacgcga aggctatcct
     3301 gtccgcgagg ctctcgaagt ccaggaggct ggagaacctg atcgctcagc tcccaggcga
     3361 gaagaagaac ggcctgttcg ggaacctcat cgctctcagc ctggggctca ccccgaactt
     3421 caagtcgaac ttcgatctcg ctgaggacgc caagctgcaa ctctccaagg acacctacga
     3481 cgatgacctc gataacctcc tggcccagat cggcgatcaa tacgcggacc tgttcctcgc
     3541 tgccaagaac ctgtcggacg ccatcctcct gtcagatatc ctccgcgtga acaccgagat
     3601 cacgaaggct ccactctctg cctccatgat caagcgctac gacgagcacc atcaggatct
     3661 gaccctcctg aaggcgctgg tccgccaaca gctcccggag aagtacaagg agattttctt
     3721 cgatcagtcg aagaacggct acgctgggta catcgacggc ggggcctcac aagaggagtt
     3781 ctacaagttc atcaagccaa tcctggagaa gatggacggc acggaggagc tcctggtgaa
     3841 gctcaacagg gaggacctcc tgcggaagca gagaaccttc gataacggca gcatccccca
     3901 ccaaatccat ctcggggagc tgcacgccat cctgagaagg caagaggact tctacccttt
     3961 cctcaaggat aaccgggaga agatcgagaa gatcctgacc ttcagaatcc catactacgt
     4021 cggccctctc gcgcggggga actcaagatt cgcttggatg acccgcaagt ctgaggagac
     4081 catcacgccg tggaacttcg aggaggtggt ggacaagggc gctagcgctc agtcgttcat
     4141 cgagaggatg accaacttcg acaagaacct gcccaacgag aaggtgctcc ctaagcactc
     4201 gctcctgtac gagtacttca ccgtctacaa cgagctcacg aaggtgaagt acgtcaccga
     4261 gggcatgcgc aagccagcgt tcctgtccgg ggagcagaag aaggctatcg tggacctcct
     4321 gttcaagacc aaccggaagg tcacggttaa gcaactcaag gaggactact tcaagaagat
     4381 cgagtgcttc gattcggtcg agatcagcgg cgttgaggac cgcttcaacg ccagcctcgg
     4441 gacctaccac gatctcctga agatcatcaa ggataaggac ttcctggaca acgaggagaa
     4501 cgaggatatc ctggaggaca tcgtgctgac cctcacgctg ttcgaggaca gggagatgat
     4561 cgaggagcgc ctgaagacgt acgcccatct cttcgatgac aaggtcatga agcaactcaa
     4621 gcgccggaga tacaccggct gggggaggct gtcccgcaag ctcatcaacg gcatccggga
     4681 caagcagtcc gggaagacca tcctcgactt cctcaagagc gatggcttcg ccaacaggaa
     4741 cttcatgcaa ctgatccacg atgacagcct caccttcaag gaggatatcc aaaaggctca
     4801 agtgagcggc cagggggact cgctgcacga gcatatcgcg aacctcgctg gctcccccgc
     4861 gatcaagaag ggcatcctcc agaccgtgaa ggttgtggac gagctcgtga aggtcatggg
     4921 ccggcacaag cctgagaaca tcgtcatcga gatggccaga gagaaccaaa ccacgcagaa
     4981 ggggcaaaag aactctaggg agcgcatgaa gcgcatcgag gagggcatca aggagctggg
     5041 gtcccaaatc ctcaaggagc acccagtgga gaacacccaa ctgcagaacg agaagctcta
     5101 cctgtactac ctccagaacg gcagggatat gtacgtggac caagagctgg atatcaaccg
     5161 cctcagcgat tacgacgtcg atcatatcgt tccccagtct ttcctgaagg atgactccat
     5221 cgacaacaag gtcctcacca ggtcggacaa gaaccgcggc aagtcagata acgttccatc
     5281 tgaggaggtc gttaagaaga tgaagaacta ctggaggcag ctcctgaacg ccaagctgat
     5341 cacgcaaagg aagttcgaca acctcaccaa ggctgagaga ggcgggctct cagagctgga
     5401 caaggccggc ttcatcaagc ggcagctggt cgagaccaga caaatcacga agcacgttgc
     5461 gcaaatcctc gactctcgga tgaacacgaa gtacgataag aacgacaagc tgatcaggga
     5521 ggttaaggtg atcaccctga agtctaagct cgtctccgac ttcaggaagg atttccagtt
     5581 ctacaaggtt cgcgagatca acaactacca ccatgcccat gacgcttacc tcaacgctgt
     5641 ggtcggcacc gctctgatca agaagtaccc aaagctggag tccgagttcg tgtacgggga
     5701 ctacaaggtt tacgatgtgc gcaagatgat cgccaagtcg gagcaagaga tcggcaaggc
     5761 taccgccaag tacttcttct actcaaacat catgaacttc ttcaagaccg agatcacgct
     5821 ggccaacggc gagatccgga agagaccgct catcgagacc aacggcgaga cgggggagat
     5881 cgtgtgggac aagggcaggg atttcgcgac cgtccgcaag gttctctcca tgccccaggt
     5941 gaacatcgtc aagaagaccg aggtccaaac gggcgggttc tcaaaggagt ctatcctgcc
     6001 taagcggaac agcgacaagc tcatcgccag aaagaaggac tgggacccaa agaagtacgg
     6061 cgggttcgac agccctaccg tggcctactc ggtcctggtt gtggcgaagg ttgagaaggg
     6121 caagtccaag aagctcaaga gcgtgaagga gctcctgggg atcaccatca tggagaggtc
     6181 cagcttcgag aagaacccaa tcgacttcct ggaggccaag ggctacaagg aggtgaagaa
     6241 ggacctgatc atcaagctcc cgaagtactc tctcttcgag ctggagaacg gcaggaagag
     6301 aatgctggct tccgctggcg agctccagaa ggggaacgag ctcgcgctgc caagcaagta
     6361 cgtgaacttc ctctacctgg cttcccacta cgagaagctc aagggcagcc cggaggacaa
     6421 cgagcaaaag cagctgttcg tcgagcagca caagcattac ctcgacgaga tcatcgagca
     6481 aatctccgag ttcagcaagc gcgtgatcct cgccgacgcg aacctggata aggtcctctc
     6541 cgcctacaac aagcaccggg acaagcccat cagagagcaa gcggagaaca tcatccatct
     6601 cttcaccctg acgaacctcg gcgctcctgc tgctttcaag tacttcgaca ccacgatcga
     6661 tcggaagaga tacacctcca cgaaggaggt cctggacgcg accctcatcc accagtcgat
     6721 caccggcctg tacgagacga ggatcgacct ctcacaactc ggcggggata agagacccgc
     6781 agcaaccaag aaggcagggc aagcaaagaa gaagaagacg cgtgactccg gcggcagcac
     6841 caacctgtcc gacatcatcg agaaggagac gggcaagcaa ctcgtgatcc aggagagcat
     6901 cctcatgctg ccagaggagg tggaggaggt catcggcaac aagccagagt ccgacatcct
     6961 ggtgcacacc gcctacgacg agtccaccga cgagaacgtc atgctcctga ccagcgacgc
     7021 cccagagtac aagccatggg ccctcgtcat ccaggacagc aacggggaga acaagatcaa
     7081 gatgctgtcg ggggggagcc caaagaagaa gcggaaggtg tagtgtacaa gtaacccggg
     7141 aattcggtac gctgaaatca ccagtctctc tctacaaatc tatctctctc tattttctcc
     7201 ataaataatg tgtgagtagt ttcccgataa gggaaattag ggttcttata gggtttcgct
     7261 catgtgttga gcatataaga aacccttagt atgtatttgt atttgtaaaa tacttctatc
     7321 aataaaattt ctaattccta aaaccaaaat ccagtactaa aatccagatc tcctaaagtc
     7381 cctatagatc tttgtcgtga atataaacca gacacgagac gactaaacct ggagcccaga
     7441 cgccgttcga agctagaagt accgcttagg caggaggccg ttagggaaaa gatgctaagg
     7501 cagggttggt tacgttgact cccccgtagg tttggtttaa atatgatgaa gtggacggaa
     7561 ggaaggagga agacaaggaa ggataaggtt gcaggccctg tgcaaggtaa gaagatggaa
     7621 atttgataga ggtacgctac tatacttata ctatacgcta agggaatgct tgtatttata
     7681 ccctataccc cctaataacc ccttatcaat ttaagaaata atccgcataa gcccccgctt
     7741 aaaaattggt atcagagcca tgaataggtc tatgaccaaa actcaagagg ataaaacctc
     7801 accaaaatac gaaagagttc ttaactctaa agataaaaga tctttcaaga tcaaaactag
     7861 ttccctcaca ccggtgacgg ggatcgcatg cgatatctcg agatctagct tggcgtaatc
     7921 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     7981 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     8041 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     8101 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     8161 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     8221 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     8281 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     8341 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     8401 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     8461 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     8521 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     8581 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     8641 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     8701 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     8761 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     8821 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     8881 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     8941 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     9001 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     9061 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     9121 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     9181 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     9241 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     9301 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     9361 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     9421 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     9481 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     9541 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     9601 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     9661 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     9721 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     9781 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     9841 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     9901 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     9961 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
    10021 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgackt
//
