LOCUS       sequence-371173-        7356 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding Nme2ABE8e driven by the CMV promoter for plasmid 
            transfection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7356)
  AUTHORS   Davis JR, Wang X, Witte IP, Huang TP, Levy JM, Raguram A, Banskota 
            S, Seidah NG, Musunuru K, Liu DR
  TITLE     Efficient in vivo base editing via single adeno-associated viruses 
            with size-optimized genomes encoding compact adenine base editors.
  JOURNAL   Nat Biomed Eng. 2022 Jul 28. pii: 10.1038/s41551-022-00911-4. doi: 
            10.1038/s41551-022-00911-4.
  PUBMED    35902773
REFERENCE   2  (bases 1 to 7356)
  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 Biomed 
            Eng. 2022 Jul 28. pii: 10.1038/s41551-022-00911-4. doi: 
            10.1038/s41551-022-00911-4."
COMMENT     Sequence Label: CMV-Nme2ABE8e
FEATURES             Location/Qualifiers
     source          1..7356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        132..335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     286..306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     377..396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             445..465
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4342..4362
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4371..4388
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4411..4428)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4417..4641
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4712..4728)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4712..4728)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4725..4747)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4736..4752
                     /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(4760..4766,4767..4784,4785..4790))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4805..4826
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4943..4960)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5114..5702)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5194..5213)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5873..6664,6665..6733))
                     /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     6496..6515
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6734..6838)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6917..6936)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(7108..7356,1..131)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat gaaacggaca
      421 gccgacggaa gcgagttcga gtcaccaaag aagaagcgga aagtctctga ggtggagttt
      481 tcccacgagt actggatgag acatgccctg accctggcca agagggcacg ggatgagagg
      541 gaggtgcctg tgggagccgt gctggtgctg aacaatagag tgatcggcga gggctggaac
      601 agagccatcg gcctgcacga cccaacagcc catgccgaaa ttatggccct gagacagggc
      661 ggcctggtca tgcagaacta cagactgatt gacgccaccc tgtacgtgac attcgagcct
      721 tgcgtgatgt gcgccggcgc catgatccac tctaggatcg gccgcgtggt gtttggcgtg
      781 aggaactcaa aaagaggcgc cgcaggctcc ctgatgaacg tgctgaacta ccccggcatg
      841 aatcaccgcg tcgaaattac cgagggaatc ctggcagatg aatgtgccgc cctgctgtgc
      901 gatttctatc ggatgcctag acaggtgttc aatgctcaga agaaggccca gagctccatc
      961 aactctggag gatctagcgg aggatcctct ggcagcgaga caccaggaac aagcgagtca
     1021 gcaacaccag agagcagtgg cggcagcagc ggcggcagcg cagcattcaa gcctaaccca
     1081 atcaattaca tcctgggact ggcaatcgga atcgcatccg tgggatgggc tatggtggag
     1141 atcgacgagg aggagaatcc tatccggctg atcgatctgg gcgtgagagt gtttgagagg
     1201 gccgaggtgc caaagaccgg cgattctctg gctatggccc ggagactggc acggagcgtg
     1261 aggcgcctga cacggagaag ggcacacagg ctgctgaggg cacgccggct gctgaagaga
     1321 gagggcgtgc tgcaggcagc agacttcgat gagaatggcc tgatcaagag cctgccaaac
     1381 accccctggc agctgagagc agccgccctg gacaggaagc tgacaccact ggagtggtct
     1441 gccgtgctgc tgcacctgat caagcaccgc ggctacctga gccagcggaa gaacgaggga
     1501 gagacagcag acaaggagct gggcgccctg ctgaagggag tggccaacaa tgcccacgcc
     1561 ctgcagaccg gcgatttcag gacacctgcc gagctggccc tgaataagtt tgagaaggag
     1621 tccggccaca tcagaaacca gaggggcgac tatagccaca ccttctcccg caaggatctg
     1681 caggccgagc tgatcctgct gttcgagaag cagaaggagt ttggcaatcc acacgtgagc
     1741 ggaggcctga aggagggaat cgagaccctg ctgatgacac agaggcctgc cctgtccggc
     1801 gacgcagtgc agaagatgct gggacactgc accttcgagc ctgcagagcc aaaggccgcc
     1861 aagaacacct acacagccga gcggtttatc tggctgacaa agctgaacaa tctgagaatc
     1921 ctggagcagg gatccgagag gccactgacc gacacagaga gggccaccct gatggatgag
     1981 ccttaccgga agtctaagct gacatatgcc caggccagaa agctgctggg cctggaggac
     2041 accgccttct ttaagggcct gagatacggc aaggataatg ccgaggcctc cacactgatg
     2101 gagatgaagg cctatcacgc catctctcgc gccctggaga aggagggcct gaaggacaag
     2161 aagtcccccc tgaacctgag ctccgagctg caggatgaga tcggcaccgc cttctctctg
     2221 tttaagaccg acgaggatat cacaggccgc ctgaaggaca gggtgcagcc tgagatcctg
     2281 gaggccctgc tgaagcacat ctctttcgat aagtttgtgc agatcagcct gaaggccctg
     2341 agaaggatcg tgccactgat ggagcagggc aagcggtacg acgaggcctg cgccgagatc
     2401 tacggcgatc actatggcaa gaagaacaca gaggagaaga tctatctgcc ccctatccct
     2461 gccgacgaga tcagaaatcc tgtggtgctg agggccctgt cccaggcaag aaaagtgatc
     2521 aacggagtgg tgcgccggta cggatctcca gcccggatcc acatcgagac cgccagagaa
     2581 gtgggcaaga gcttcaagga ccggaaggag atcgagaaga gacaggagga gaatcgcaag
     2641 gatcgggaga aggccgccgc caagtttagg gagtacttcc ctaactttgt gggcgagcca
     2701 aagtctaagg acatcctgaa gctgcgcctg tacgagcagc agcacggcaa gtgtctgtat
     2761 agcggcaagg agatcaatct ggtgcggctg aacgagaagg gctatgtgga gatcgatcac
     2821 gccctgcctt tctccagaac ctgggacgat tcttttaaca ataaggtgct ggtgctgggc
     2881 agcgagaacc agaataaggg caatcagaca ccatacgagt atttcaatgg caaggacaac
     2941 tccagggagt ggcaggagtt caaggcccgc gtggagacct ctagatttcc caggagcaag
     3001 aagcagcgga tcctgctgca gaagttcgac gaggatggct ttaaggagtg caacctgaat
     3061 gacaccagat acgtgaaccg gttcctgtgc cagtttgtgg ccgatcacat cctgctgacc
     3121 ggcaagggca agagaagggt gttcgcctct aatggccaga tcacaaacct gctgagggga
     3181 ttttggggac tgaggaaggt gcgggcagag aatgacagac accacgcact ggatgcagtg
     3241 gtggtggcat gcagcaccgt ggcaatgcag cagaagatca caagattcgt gaggtataag
     3301 gagatgaacg cctttgacgg caagaccatc gataaggaga caggcaaggt gctgcaccag
     3361 aagacccact tcccccagcc ttgggagttc tttgcccagg aagtgatgat ccgggtgttc
     3421 ggcaagccag acggcaagcc tgagtttgag gaggccgata ccccagagaa gctgaggaca
     3481 ctgctggcag agaagctgtc tagcaggcca gaggcagtgc acgagtacgt gaccccactg
     3541 ttcgtgtcca gggcacccaa tcggaagatg tctggcgccc acaaggacac actgagaagc
     3601 gccaagaggt ttgtgaagca caacgagaag atctccgtga agagagtgtg gctgaccgag
     3661 atcaagctgg ccgatctgga gaacatggtg aattacaaga acggcaggga gatcgagctg
     3721 tatgaggccc tgaaggcaag gctggaggcc tacggaggaa atgccaagca ggccttcgac
     3781 ccaaaggata acccctttta taagaaggga ggacagctgg tgaaggccgt gcgggtggag
     3841 aagacccagg agagcggcgt gctgctgaat aagaagaacg cctacacaat cgccgacaat
     3901 ggcgatatgg tgagagtgga cgtgttctgt aaggtggata agaagggcaa gaatcagtac
     3961 tttatcgtgc ctatctatgc ctggcaggtg gccgagaaca tcctgccaga catcgattgc
     4021 aagggctaca gaatcgacga tagctataca ttctgttttt ccctgcacaa gtatgacctg
     4081 atcgccttcc agaaggatga gaagtccaag gtggagtttg cctactatat caattgcgac
     4141 tcctctaacg gcaggttcta cctggcctgg cacgataagg gcagcaagga gcagcagttt
     4201 cgcatctcca cccagaatct ggtgctgatc cagaagtatc aggtgaacga gctgggcaag
     4261 gagatcaggc catgtcggct gaagaagcgc ccacccgtgt ctggcggctc aaaaagaacc
     4321 gccgacggca gcgaattcga gcccaagaag aagaggaaag tctaaccggt catcatcacc
     4381 atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     4441 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     4501 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     4561 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     4621 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tcgataccgt
     4681 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4741 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctagggtg
     4801 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4861 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4921 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4981 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5041 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5101 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5161 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5221 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5281 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5341 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5401 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5461 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5521 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5581 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5641 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5701 aagaagatcc tttgatcttt tctacggggt ctgacactca gtggaacgaa aactcacgtt
     5761 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5821 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5881 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5941 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6001 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6061 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6121 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6181 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6241 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6301 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6361 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6421 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6481 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6541 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6601 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6661 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6721 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6781 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6841 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcgatct cccgatcccc
     6901 tagggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     6961 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     7021 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
     7081 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
     7141 caattacggg gtcattagtt catagcccat atattgagtt ccgcgttaca taacttacgg
     7201 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     7261 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     7321 ggtaaactgc ccacttggca gtacatcaag tgtatc
//
