LOCUS       sequence-257653-        8827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Conversion of C to T in potato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8827)
  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 8827)
  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: p35S-A3A-PBE
FEATURES             Location/Qualifiers
     source          1..8827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        401..746
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     689..709
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             5521..5568
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5590..5838
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5851..5871
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5945..6121
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(6675..6691)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6675..6691)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6688..6710)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6699..6715
                     /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(6723..6729,6730..6747,6748..6753))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6768..6789
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6906..6923)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7077..7665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7157..7176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7836..8627,8628..8696))
                     /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     8459..8478
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8697..8801)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gagctcggta cccctactcc aaaaatgtca aagatacagt ctcagaagac caaagggcta
       61 ttgagacttt tcaacaaagg gtaatttcgg gaaacctcct cggattccat tgcccagcta
      121 tctgtcactt catcgaaagg acagtagaaa aggaaggtgg ctcctacaaa tgccatcatt
      181 gcgataaagg aaaggctatc attcaagatg cctctgccga cagtggtccc aaagatggac
      241 ccccacccac gaggagcatc gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag
      301 tggattgatg tgacatctcc actgacgtaa gggatgacgc acaatcccac ccctactcca
      361 aaaatgtcaa agatacagtc tcagaagacc aaagggctat tgagactttt caacaaaggg
      421 taatttcggg aaacctcctc ggattccatt gcccagctat ctgtcacttc atcgaaagga
      481 cagtagaaaa ggaaggtggc tcctacaaat gccatcattg cgataaagga aaggctatca
      541 ttcaagatgc ctctgccgac agtggtccca aagatggacc cccacccacg aggagcatcg
      601 tggaaaaaga agacgttcca accacgtctt caaagcaagt ggattgatgt gacatctcca
      661 ctgacgtaag ggatgacgca caatcccact atccttcgca agacccttcc tctatataag
      721 gaagttcatt tcatttggag aggacagccc aagcttgtcg acggatccat ggaggccagc
      781 ccggctagcg gcccaaggca tctcatggac ccgcacatct tcaccagcaa cttcaacaac
      841 ggcatcggca ggcacaagac ctacttgtgc tacgaggtgg agaggctcga caacggaacc
      901 tccgtgaaga tggaccaaca cagggggttc ctccacaacc aagccaagaa cctcctctgc
      961 ggcttctacg gcaggcacgc cgagttgagg ttcctcgact tggtgccatc cctccaactc
     1021 gatccagccc aaatctaccg cgtgacctgg ttcatctcct ggtccccatg cttctcctgg
     1081 ggttgcgccg gcgaggttcg ggctttcctc caagaaaaca cccacgtccg cctccgcatt
     1141 ttcgccgcca ggatctatga ttacgaccct ctctacaagg aggccctcca gatgctgcgg
     1201 gacgccggtg ctcaggtgag tatcatgacc tacgacgagt tcaagcactg ctgggacacc
     1261 ttcgttgacc accagggctg cccattccaa ccatgggacg gtctggatga acacagccaa
     1321 gccttgtccg gcaggctccg ggccatcctc caaaaccagg ggaactccgg gagcgagacg
     1381 ccaggcacct ccgagtcggc caccccagaa tctcttaagg acaagaagta ctcgatcggc
     1441 ctcgccatcg ggacgaactc agttggctgg gccgtgatca ccgacgagta caaggtgccc
     1501 tctaagaagt tcaaggtcct ggggaacacc gaccgccatt ccatcaagaa gaacctcatc
     1561 ggcgctctcc tgttcgacag cggggagacc gctgaggcta cgaggctcaa gagaaccgct
     1621 aggcgccggt acacgagaag gaagaacagg atctgctacc tccaagagat tttctccaac
     1681 gagatggcca aggttgacga ttcattcttc caccgcctgg aggagtcttt cctcgtggag
     1741 gaggataaga agcacgagcg gcatcccatc ttcggcaaca tcgtggacga ggttgcctac
     1801 cacgagaagt accctacgat ctaccatctg cggaagaagc tcgtggactc caccgataag
     1861 gcggacctca gactgatcta cctcgctctg gcccacatga tcaagttccg cggccatttc
     1921 ctgatcgagg gggatctcaa cccagacaac agcgatgttg acaagctgtt catccaactc
     1981 gtgcagacct acaaccaact cttcgaggag aacccgatca acgcctctgg cgtggacgcg
     2041 aaggctatcc tgtccgcgag gctctcgaag tccaggaggc tggagaacct gatcgctcag
     2101 ctcccaggcg agaagaagaa cggcctgttc gggaacctca tcgctctcag cctggggctc
     2161 accccgaact tcaagtcgaa cttcgatctc gctgaggacg ccaagctgca actctccaag
     2221 gacacctacg acgatgacct cgataacctc ctggcccaga tcggcgatca atacgcggac
     2281 ctgttcctcg ctgccaagaa cctgtcggac gccatcctcc tgtcagatat cctccgcgtg
     2341 aacaccgaga tcacgaaggc tccactctct gcctccatga tcaagcgcta cgacgagcac
     2401 catcaggatc tgaccctcct gaaggcgctg gtccgccaac agctcccgga gaagtacaag
     2461 gagattttct tcgatcagtc gaagaacggc tacgctgggt acatcgacgg cggggcctca
     2521 caagaggagt tctacaagtt catcaagcca atcctggaga agatggacgg cacggaggag
     2581 ctcctggtga agctcaacag ggaggacctc ctgcggaagc agagaacctt cgataacggc
     2641 agcatccccc accaaatcca tctcggggag ctgcacgcca tcctgagaag gcaagaggac
     2701 ttctaccctt tcctcaagga taaccgggag aagatcgaga agatcctgac cttcagaatc
     2761 ccatactacg tcggccctct cgcgcggggg aactcaagat tcgcttggat gacccgcaag
     2821 tctgaggaga ccatcacgcc gtggaacttc gaggaggtgg tggacaaggg cgctagcgct
     2881 cagtcgttca tcgagaggat gaccaacttc gacaagaacc tgcccaacga gaaggtgctc
     2941 cctaagcact cgctcctgta cgagtacttc accgtctaca acgagctcac gaaggtgaag
     3001 tacgtcaccg agggcatgcg caagccagcg ttcctgtccg gggagcagaa gaaggctatc
     3061 gtggacctcc tgttcaagac caaccggaag gtcacggtta agcaactcaa ggaggactac
     3121 ttcaagaaga tcgagtgctt cgattcggtc gagatcagcg gcgttgagga ccgcttcaac
     3181 gccagcctcg ggacctacca cgatctcctg aagatcatca aggataagga cttcctggac
     3241 aacgaggaga acgaggatat cctggaggac atcgtgctga ccctcacgct gttcgaggac
     3301 agggagatga tcgaggagcg cctgaagacg tacgcccatc tcttcgatga caaggtcatg
     3361 aagcaactca agcgccggag atacaccggc tgggggaggc tgtcccgcaa gctcatcaac
     3421 ggcatccggg acaagcagtc cgggaagacc atcctcgact tcctcaagag cgatggcttc
     3481 gccaacagga acttcatgca actgatccac gatgacagcc tcaccttcaa ggaggatatc
     3541 caaaaggctc aagtgagcgg ccagggggac tcgctgcacg agcatatcgc gaacctcgct
     3601 ggctcccccg cgatcaagaa gggcatcctc cagaccgtga aggttgtgga cgagctcgtg
     3661 aaggtcatgg gccggcacaa gcctgagaac atcgtcatcg agatggccag agagaaccaa
     3721 accacgcaga aggggcaaaa gaactctagg gagcgcatga agcgcatcga ggagggcatc
     3781 aaggagctgg ggtcccaaat cctcaaggag cacccagtgg agaacaccca actgcagaac
     3841 gagaagctct acctgtacta cctccagaac ggcagggata tgtacgtgga ccaagagctg
     3901 gatatcaacc gcctcagcga ttacgacgtc gatcatatcg ttccccagtc tttcctgaag
     3961 gatgactcca tcgacaacaa ggtcctcacc aggtcggaca agaaccgcgg caagtcagat
     4021 aacgttccat ctgaggaggt cgttaagaag atgaagaact actggaggca gctcctgaac
     4081 gccaagctga tcacgcaaag gaagttcgac aacctcacca aggctgagag aggcgggctc
     4141 tcagagctgg acaaggccgg cttcatcaag cggcagctgg tcgagaccag acaaatcacg
     4201 aagcacgttg cgcaaatcct cgactctcgg atgaacacga agtacgatga gaacgacaag
     4261 ctgatcaggg aggttaaggt gatcaccctg aagtctaagc tcgtctccga cttcaggaag
     4321 gatttccagt tctacaaggt tcgcgagatc aacaactacc accatgccca tgacgcttac
     4381 ctcaacgctg tggtcggcac cgctctgatc aagaagtacc caaagctgga gtccgagttc
     4441 gtgtacgggg actacaaggt ttacgatgtg cgcaagatga tcgccaagtc ggagcaagag
     4501 atcggcaagg ctaccgccaa gtacttcttc tactcaaaca tcatgaactt cttcaagacc
     4561 gagatcacgc tggccaacgg cgagatccgg aagagaccgc tcatcgagac caacggcgag
     4621 acgggggaga tcgtgtggga caagggcagg gatttcgcga ccgtccgcaa ggttctctcc
     4681 atgccccagg tgaacatcgt caagaagacc gaggtccaaa cgggcgggtt ctcaaaggag
     4741 tctatcctgc ctaagcggaa cagcgacaag ctcatcgcca gaaagaagga ctgggaccca
     4801 aagaagtacg gcgggttcga cagccctacc gtggcctact cggtcctggt tgtggcgaag
     4861 gttgagaagg gcaagtccaa gaagctcaag agcgtgaagg agctcctggg gatcaccatc
     4921 atggagaggt ccagcttcga gaagaaccca atcgacttcc tggaggccaa gggctacaag
     4981 gaggtgaaga aggacctgat catcaagctc ccgaagtact ctctcttcga gctggagaac
     5041 ggcaggaaga gaatgctggc ttccgctggc gagctccaga aggggaacga gctcgcgctg
     5101 ccaagcaagt acgtgaactt cctctacctg gcttcccact acgagaagct caagggcagc
     5161 ccggaggaca acgagcaaaa gcagctgttc gtcgagcagc acaagcatta cctcgacgag
     5221 atcatcgagc aaatctccga gttcagcaag cgcgtgatcc tcgccgacgc gaacctggat
     5281 aaggtcctct ccgcctacaa caagcaccgg gacaagccca tcagagagca agcggagaac
     5341 atcatccatc tcttcaccct gacgaacctc ggcgctcctg ctgctttcaa gtacttcgac
     5401 accacgatcg atcggaagag atacacctcc acgaagaagg tcctggacgc gaccctcatc
     5461 caccagtcga tcaccggcct gtacgagacg aggatcgacc tctcacaact cggcggggat
     5521 aagagacccg cagcaaccaa gaaggcaggg caagcaaaga agaagaagac gcgtgactcc
     5581 ggcggcagca ccaacctgtc cgacatcatc gagaaggaga cgggcaagca actcgtgatc
     5641 caggagagca tcctcatgct gccagaggag gtggaggagg tcatcggcaa caagccagag
     5701 tccgacatcc tggtgcacac cgcctacgac gagtccaccg acgagaacgt catgctcctg
     5761 accagcgacg ccccagagta caagccatgg gccctcgtca tccaggacag caacggggag
     5821 aacaagatca agatgctgtc gggggggagc ccaaagaaga agcggaaggt gtagtgtaca
     5881 agtaacccgg gaattcggta cgctgaaatc accagtctct ctctacaaat ctatctctct
     5941 ctattttctc cataaataat gtgtgagtag tttcccgata agggaaatta gggttcttat
     6001 agggtttcgc tcatgtgttg agcatataag aaacccttag tatgtatttg tatttgtaaa
     6061 atacttctat caataaaatt tctaattcct aaaaccaaaa tccagtacta aaatccagat
     6121 ctcctaaagt ccctatagat ctttgtcgtg aatataaacc agacacgaga cgactaaacc
     6181 tggagcccag acgccgttcg aagctagaag taccgcttag gcaggaggcc gttagggaaa
     6241 agatgctaag gcagggttgg ttacgttgac tcccccgtag gtttggttta aatatgatga
     6301 agtggacgga aggaaggagg aagacaagga aggataaggt tgcaggccct gtgcaaggta
     6361 agaagatgga aatttgatag aggtacgcta ctatacttat actatacgct aagggaatgc
     6421 ttgtatttat accctatacc ccctaataac cccttatcaa tttaagaaat aatccgcata
     6481 agcccccgct taaaaattgg tatcagagcc atgaataggt ctatgaccaa aactcaagag
     6541 gataaaacct caccaaaata cgaaagagtt cttaactcta aagataaaag atctttcaag
     6601 atcaaaacta gttccctcac accggtgacg gggatcgcat gcgatatctc gagatctagc
     6661 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     6721 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6781 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6841 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     6901 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6961 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     7021 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7081 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7141 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     7201 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     7261 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     7321 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7381 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7441 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7501 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7561 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     7621 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     7681 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     7741 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     7801 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7861 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     7921 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     7981 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     8041 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     8101 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     8161 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     8221 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     8281 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     8341 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     8401 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     8461 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     8521 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     8581 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     8641 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     8701 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     8761 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     8821 ccacctg
//
