LOCUS       sequence-371172-        7065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding CjABE8e driven by the CMV promoter for plasmid 
            transfection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7065)
  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 7065)
  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-CjABE8e
FEATURES             Location/Qualifiers
     source          1..7065
                     /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             4051..4071
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4080..4097
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4120..4137)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4126..4350
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4421..4437)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4421..4437)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4434..4456)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4445..4461
                     /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(4469..4475,4476..4493,4494..4499))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4514..4535
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4652..4669)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4823..5411)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4903..4922)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5582..6373,6374..6442))
                     /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     6205..6224
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6443..6547)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6626..6645)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(6817..7065,1..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 cccgcatcct cgctttcgca
     1081 atcggaatct ctagtatcgg atgggccttc tctgaaaacg acgaactgaa agactgcggc
     1141 gtgagaatct tcacaaaggt tgaaaaccct aaaacaggcg agtctttagc tctgccacgt
     1201 aggttggccc gctccgcccg aaaaaggctg gctcggcgga aggctcgcct caaccacttg
     1261 aagcatttga tagctaatga gttcaaactg aactacgaag attaccagtc cttcgacgag
     1321 tcattggcaa aagcctacaa aggcagcctt atcagtcctt atgagttgag atttcgcgca
     1381 ctcaacgaac tgctttctaa gcaagacttt gctagggtca ttctgcacat cgcaaaacgg
     1441 cgaggttatg acgatatcaa gaactccgac gataaagaaa agggagccat tctcaaggcg
     1501 atcaaacaga atgaggaaaa attggcaaac taccagagtg tgggcgagta tctgtataaa
     1561 gagtatttcc agaagtttaa ggaaaacagc aaggagttta caaacgtcag aaataaaaag
     1621 gagtcttacg agagatgcat cgcgcagtca ttcctcaaag atgagctgaa gctgatattt
     1681 aagaagcaac gcgaatttgg tttctcattc tctaagaagt tcgaagagga ggttctttcc
     1741 gtggcgtttt acaagagggc gctcaaagac ttctcccacc tggttggtaa ctgtagtttc
     1801 ttcacggatg agaagcgagc tcccaaaaat tctcccctgg ctttcatgtt tgttgccctg
     1861 actcggatca ttaacctgct gaacaacctg aaaaatactg aagggatctt gtatacgaag
     1921 gacgacctaa atgcactcct gaatgaagtg ctcaaaaacg gaactctaac ctataaacag
     1981 accaagaaat tactggggct ctctgacgac tacgagttca agggcgagaa gggtacttat
     2041 tttatcgaat tcaaaaagta taaggagttc attaaagcat tgggggaaca caacctcagc
     2101 caggacgatc tcaatgaaat tgccaaggac atcacgctga ttaaagacga gataaaactg
     2161 aaaaaggcac tggccaagta tgacctcaac cagaaccaga tcgactctct gtccaagctg
     2221 gagttcaaag accacctaaa catatccttc aaagccctga aactggtcac ccctctaatg
     2281 ctcgaaggaa aaaaatacga cgaggcgtgt aatgaactga atcttaaggt ggccatcaat
     2341 gaggataaga aggactttct tccagccttt aacgagacat attacaaaga cgaggtcaca
     2401 aacccggttg tgctgagggc cataaaagag tatcggaagg ttctgaatgc cctcctgaag
     2461 aagtacggca aagtgcacaa aataaatatc gaattggcta gggaggtggg gaagaaccat
     2521 tctcagcgag caaagatcga gaaagagcag aatgagaact acaaagccaa gaaagacgcc
     2581 gaactggagt gcgaaaagct ggggcttaaa ataaacagta aaaacatcct gaaattaaga
     2641 ttgttcaaag agcaaaagga gttttgcgcc tactcagggg aaaaaatcaa aatatcagac
     2701 ctgcaggacg agaaaatgct ggagatcgac cacatctatc cgtatagcag gtcatttgac
     2761 gattcctaca tgaacaaagt gcttgtgttt accaaacaga accaagaaaa gctgaaccaa
     2821 accccctttg aggctttcgg aaacgactca gccaagtggc agaaaatcga agtcctagcc
     2881 aagaatctgc ctacaaaaaa acaaaagagg attcttgata agaactataa ggacaaggaa
     2941 cagaaaaact ttaaagacag gaacctgaat gacacgaggt acattgcgcg actggttcta
     3001 aactatacca aagactacct ggatttcctc cctctgagcg acgacgagaa tactaaactg
     3061 aatgataccc agaaaggctc aaaggtccac gttgaggcta agtccgggat gctgactagc
     3121 gccctccgcc acacgtgggg cttcagcgcc aaagatcgga ataatcatct tcatcacgct
     3181 attgatgcag taatcatagc ctacgctaac aacagcatcg tgaaagcctt ctccgatttc
     3241 aagaaagaac aggagtctaa tagcgccgag ttgtacgcca agaaaatttc cgaattggac
     3301 tataaaaata agagaaaatt cttcgaaccc ttctccgggt ttcgccaaaa ggtcttagat
     3361 aagatcgacg agattttcgt ttccaagccc gaaagaaaaa agccttcagg ggcactgcac
     3421 gaagagacat tccgcaagga agaggaattt taccaatctt acggtggtaa agagggagtt
     3481 ctgaaggctc tggagcttgg gaagatccgc aaggtaaacg ggaaaatcgt gaaaaacggg
     3541 gacatgttca gggtggatat cttcaagcac aaaaagacca acaagttcta cgcagtaccc
     3601 atctacacta tggatttcgc tttaaaggtt ctcccaaata aggcggtggc tcgatcgaag
     3661 aaaggagaga tcaaggactg gatcttaatg gatgaaaatt acgagttttg cttctcgctc
     3721 tacaaagata gcctgattct gatccagaca aaagacatgc aggaaccaga atttgtttat
     3781 tataacgcct tcacgagcag tacagtgtcc ctgattgtga gcaagcatga taacaagttc
     3841 gagactctgt ctaagaatca gaaaatcctt ttcaagaacg ccaacgagaa ggaggtcatc
     3901 gcaaagtcaa ttggcatcca aaacctgaag gtgttcgaga aatacatagt gtccgcactc
     3961 ggtgaagtaa ctaaagccga atttcgacag cgcgaggatt ttaagaaatc tggcggctca
     4021 aaaagaaccg ccgacggcag cgaattcgag cccaagaaga agaggaaagt ctaaccggtc
     4081 atcatcacca tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     4141 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     4201 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     4261 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     4321 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     4381 cgataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4441 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4501 cctagggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4561 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4621 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4681 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4741 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4801 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4861 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4921 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4981 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5041 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5101 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5161 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5221 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5281 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5341 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5401 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacactcag tggaacgaaa
     5461 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5521 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5581 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5641 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5701 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5761 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5821 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5881 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5941 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6001 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6061 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6121 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6181 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6241 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6301 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6361 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6421 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6481 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6541 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatcgatctc
     6601 ccgatcccct agggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     6661 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     6721 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
     6781 cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt
     6841 aatagtaatc aattacgggg tcattagttc atagcccata tattgagttc cgcgttacat
     6901 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     6961 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     7021 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatc
//
