LOCUS       sequence-351811-        7238 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression  of base editor in mammalian cells .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7238)
  AUTHORS   Mok BY, Kotrys AV, Raguram A, Huang TP, Mootha VK, Liu DR
  TITLE     CRISPR-free base editors with enhanced activity and expanded 
            targeting scope in mitochondrial and nuclear DNA.
  JOURNAL   Nat Biotechnol. 2022 Sep;40(9):1378-1387. doi: 
            10.1038/s41587-022-01256-8. Epub 2022 Apr 4.
  PUBMED    35379961
REFERENCE   2  (bases 1 to 7238)
  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"; doi: 
            "10.1038/s41587-022-01256-8"; journalName: "Nat Biotechnol"; date: 
            "2022-09"; volume: "40"; issue: "9"; pages: "1378-1387"
COMMENT     Sequence Label: ND4.2-Left TALE-G1397-N-DddA11
FEATURES             Location/Qualifiers
     source          1..7238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(107..126)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        298..677
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        678..881
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     832..852
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     926..945
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        926..944
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             967..1044
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MASVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1057..1122
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3085..3336
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     rep_origin      3533..3961
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3620..3639)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3830..3851
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3970..3990)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3975..4304
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4155..4290
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4217..4236
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4352..4399
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4418..4816
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4974..5095
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5011..5030)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5065..5084
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5144..5160)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5144..5160)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5157..5179)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5168..5184
                     /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(5192..5198,5199..5216,5217..5222))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5237..5258
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5375..5392)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5546..6134)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5626..5645)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6305..7096,7097..7165))
                     /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     6928..6947
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7166..7238,1..32))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
       61 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
      121 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      181 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      241 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      301 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      361 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      421 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      481 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      541 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      601 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      661 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      721 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      781 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      841 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac
      901 ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct
      961 agcaccatgg cctccgttct gaccccgctg ctgctgcgtg gcctgaccgg ctccgctcgt
     1021 cgtctgccag ttccgcgtgc gaaaatccat tccctggact acaaagacca tgacggtgat
     1081 tataaagatc atgacatcga ttacaaggat gacgatgaca agatggacat cgcggatctg
     1141 cgtaccctgg gttacagcca gcagcagcag gagaagatca agccgaaggt gcgcagcacc
     1201 gtggctcagc accacgaagc cctggtgggc cacggtttca cccacgctca catcgtggcc
     1261 ctgagccagc acccagccgc gctgggcacc gtggccgtga aatatcagga catgattgct
     1321 gccctgccag aggccaccca tgaagctatt gtgggcgtgg gcaagcagtg gagcggtgct
     1381 cgtgcgctgg aggcgctgct gaccgtggct ggtgaactgc gtggtccgcc gctgcagctg
     1441 gataccggtc agctgctgaa aattgccaaa cgtggcggtg tgaccgcggt ggaagccgtg
     1501 catgcttggc gtaatgctct gaccggtgcg ccgctgaact taacgccgga acaagttgtt
     1561 gcaattgcca gcaacaacgg cggcaagcaa gctctggaaa ccgtgcaggc cctgcttcca
     1621 gtcctgtgtc aggcccacgg cctgacccct cagcaggtgg tcgccatcgc ctctaacatc
     1681 ggcggaaagc aggctctgga gaccgtgcag cggctgctgc ccgtgctgtg ccaggcccac
     1741 ggcctgacac ctcagcaggt ggtggccatt gctagcaaca tcggaggcaa gcaggccctg
     1801 gaaaccgtgc aaagactgct gcccgtgctg tgccaggccc acggcctcac acccgagcaa
     1861 gtggtggcca tcgctagcaa caacggcggc aagcaggctc tcgagacagt ccaagctctg
     1921 ctgccagtgc tgtgccaagc tcacggcctg acacccgagc aggtggtcgc catcgcctcc
     1981 cacgatggcg gcaaacaggc tctggaaaca gtgcaggccc tgctgcctgt gctgtgccag
     2041 gcccacggcc tgacccctga gcaggtggtg gccatcgcca gcaatggcgg aggcaagcag
     2101 gccctggaaa ccgtgcaggc tctgctgcct gtgctgtgtc aagcccatgg cctgacccct
     2161 gagcaggtgg tggctatcgc ctctaatggc ggaggcaagc aggccctgga aaccgtgcag
     2221 agactgctgc ctgtgctgtg ccaggcccat ggactgaccc ctgagcaggt tgttgcaatc
     2281 gccagccacg acggcggcaa acaagccctg gaaaccgtgc aggccctgct gcccgtgctg
     2341 tgccaggccc acggactgac cccacagcag gttgtggcca tcgccagcaa catcggcggc
     2401 aagcaggccc tggaaacagt gcagcggctg ctgcctgtgc tgtgtcaggc ccacggcctg
     2461 acccctcaac aggtggtggc catcgccagc cacgacggcg gcaagcaggc tcttgagacc
     2521 gtgcagagac tgctgcccgt gctgtgccag gcccacggac tgacacctca gcaggtggtg
     2581 gccatcgcct cccacgacgg cgggagacct gccctcgaaa gcattgtggc tcagctgagc
     2641 cgtccagacc cggcgctggc ggctctgacc aacgatcacc tggtggcgct ggcttgcctg
     2701 ggcggtcgtc cggccctgga tgccgtgaag aaaggcctgg gtggatccgg cagctacgcc
     2761 ctgggtccgt atcagattag cgccccgcag ctgccagctt acaatggtca gaccgtgggt
     2821 accttctact atgtgaacga cgcgggcggt ctggagagca aggtgtttat cagcggcggt
     2881 ccaaccccgt acccaaacta tgtcagtgcc ggtcatgtgg agggtcagag cgccctgttc
     2941 atgcgtgata acggcatcag cgagggtctg gtgttccaca acaacccgaa aggcacctgc
     3001 ggtttttgcg tgaacatgat cgagaccctg ctgccggaaa acgcgaaaat gaccgtggtg
     3061 ccgccggaag gttctggcgg ctcaactaat ctgagcgaca tcattgagaa ggagactggg
     3121 aaacagctgg tcattcagga gtccatcctg atgctgcctg aggaggtgga ggaagtgatc
     3181 ggcaacaagc cagagtctga catcctggtg cacaccgcct acgacgagtc cacagatgag
     3241 aatgtgatgc tgctgacctc tgacgccccc gagtataagc cttgggccct ggtcatccag
     3301 gattctaacg gcgagaataa gatcaagatg ctgtgataag gggtctttgt cctctgtact
     3361 gtctctctcc ttgcccctaa cccaaaaagc ttcatttttc tgtgtaggct gcacaagagc
     3421 cttgattgaa gatatattct ttctgaacag tatttaaggt ttccaataaa atgtacaccc
     3481 ctcagaactt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     3541 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3601 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3661 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3721 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3781 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3841 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3901 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3961 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4021 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4081 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4141 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4201 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     4261 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     4321 gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat
     4381 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagcctt tgtctcaaga
     4441 agaatccacc ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga
     4501 ctacagcgtc gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt
     4561 atatcatttt actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc
     4621 ggcagctggc aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag
     4681 cccctgcgga cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt
     4741 gaaggacagt gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta
     4801 tgtgtgggag ggctaagcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt
     4861 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4921 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4981 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5041 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5101 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5161 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5221 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5281 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5341 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5401 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5461 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5521 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5581 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5641 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5701 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5761 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5821 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5881 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5941 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6001 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6061 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6121 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6181 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6241 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6301 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6361 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6421 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6481 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6541 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6601 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6661 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6721 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6781 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6841 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6901 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6961 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7021 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7081 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7141 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7201 agggttattg tctcatgagc ggatacatat ttgaatgt
//
