LOCUS       sequence-351814-        7184 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 7184)
  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 7184)
  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-Right DdCBE-G1397-C- T1413I
FEATURES             Location/Qualifiers
     source          1..7184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(2..106)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(181..200)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        372..751
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        752..955
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     906..926
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1000..1019
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1000..1018
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1134..1160
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1164..1190
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1194..1220
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3051..3302
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     rep_origin      3553..3981
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3640..3659)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3850..3871
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3990..4010)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3995..4324
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4175..4310
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4237..4256
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4372..4419
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4438..4836
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4994..5115
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5031..5050)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5085..5104
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5164..5180)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5164..5180)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5177..5199)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5188..5204
                     /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(5212..5218,5219..5236,5237..5242))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5257..5278
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5395..5412)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5566..6154)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5646..5665)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6325..7116,7117..7184,1))
                     /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     6948..6967
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
       61 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
      121 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
      181 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
      241 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
      301 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
      361 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
      421 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      481 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      541 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      601 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      661 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      721 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      781 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      841 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      901 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
      961 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
     1021 gacccaagct ggctagcacc atggccctgt cccgtgcggt ttgtggcacc tcccgtcaac
     1081 tggctccggt tctgggttat ctgggttccc gtcaaaaaca ctccctgccg gactacccgt
     1141 atgatgttcc ggattacgct ggctacccat acgacgtccc agactacgct ggctacccat
     1201 acgacgtccc agactacgct atggacatcg cggatctgcg taccctgggt tacagccagc
     1261 agcagcagga gaagatcaag ccgaaggtgc gcagcaccgt ggctcagcac cacgaagccc
     1321 tggtgggcca cggtttcacc cacgctcaca ttgtggccct gagccagcac ccagccgcgc
     1381 tgggcaccgt ggccgtgaaa tatcaggata tgattgctgc cctgccagag gccacccatg
     1441 aagctattgt gggcgtgggc aagcagtgga gcggtgctcg tgcgctggag gcgctgctga
     1501 ccgtggctgg tgaactgcgt ggtccgccgc tgcagctgga caccggtcag ctgctgaaaa
     1561 tcgcgaaacg tggcggtgtg accgcggtgg aagccgtgca tgcttggcgt aatgctctga
     1621 ccggtgcgcc gctgaacctc acacccgaac aggttgttgc tatcgccagc aacaacggag
     1681 gaaagcaggc ccttgagaca gtgcaggccc tgctccccgt gctgtgccag gcccacggac
     1741 tgacccctca acaggtggtt gctatcgcct ccaacaacgg cggcaagcag gccctggaaa
     1801 ccgtgcagag gctgctgccc gtgctgtgtc aggcccacgg cctgacccct cagcaggtcg
     1861 tggccatcgc cagcaacaac ggcggaaagc aggcacttga aaccgtccaa agactgctgc
     1921 ccgtgctgtg ccaggcccat ggactgacac ccgagcaggt ggtggccatt gccagccacg
     1981 acggcggaaa gcaagctctg gagaccgttc aagctctgct gccagtgctg tgccaggccc
     2041 acggactgac accagagcag gtggtggcta tcgcctctaa caacggcggc aagcaggccc
     2101 ttgaaaccgt gcaggctctg ctgcctgtgc tgtgccaggc tcacggcctg acacctgagc
     2161 aggtcgtggc catcgcctct aatatcggcg gcaagcaggc cctggagaca gtgcaggctc
     2221 tgctgcctgt tctgtgtcag gcccacggcc tgacccctga gcaggtggtg gccatcgcca
     2281 gcaacggcgg cggcaagcag gccctcgaga cagtgcagag actgctgccc gtgctgtgcc
     2341 aggcccacgg cctgacacct gagcaggtgg tggctatcgc cagcaacggc ggcggcaaac
     2401 aagccctgga aacagtgcaa gctcttctgc ctgtgctgtg tcaggcccac ggactgaccc
     2461 ctcagcaggt ggtggccatc gccagcaaca tcggcggtaa gcaggccctg gaaaccgtgc
     2521 agcggctgct gcctgtgctg tgccaggccc acggcctgac ccctgagcaa gtggtggcca
     2581 tcgctagcaa tggaggcggc aaacaggccc tggaaaccgt gcagcggctg ctgcctgtgc
     2641 tgtgccaggc ccacggcctg accccacaac aagtggtggc catcgccagc aacaatggcg
     2701 gcaaacaggc cctggaaacc gtgcagagac tgctgcctgt gctgtgccag gcccacggcc
     2761 tgacccccca gcaggtggtg gccatcgcct ccaatatcgg cgggagaccc gccctggaaa
     2821 gcattgtggc tcagctgagc cgtccagacc cggccctggc ggctctgacc aacgatcacc
     2881 tggtggcgct ggcttgcctg ggcggtcgtc cggccctgga tgcggtgaag aaaggcctgg
     2941 gtggatccgc cattccagtg aagcgcggcg ctaccggtga aaccaaagtg tttatcggta
     3001 acagcaacag cccgaagagc ccgaccaaag gcggttgctc tggcggctca actaatctga
     3061 gcgacatcat tgagaaggag actgggaaac agctggtcat tcaggagtcc atcctgatgc
     3121 tgcctgagga ggtggaggaa gtgatcggca acaagccaga gtctgacatc ctggtgcaca
     3181 ccgcctacga cgagtccaca gatgagaatg tgatgctgct gacctctgac gcccccgagt
     3241 ataagccttg ggccctggtc atccaggatt ctaacggcga gaataagatc aagatgctgt
     3301 gataaaccac gatcgttatg ctgatcatac cctaatgatc ccagcaagat aatgtcctgt
     3361 cttctaagat gtgcatcaag cctggtacat actgaaaacc ctataaggtc ctggataatt
     3421 tttgtttgat tattcattga agaaacattt attttccaat tgtgtgaagt ttttgactgt
     3481 taataaaaga atctgtcaac catcaaactt ctgaggcgga aagaaccagc tggggctcta
     3541 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3601 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3661 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3721 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3781 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3841 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3901 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3961 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4021 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4081 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4141 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4201 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4261 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4321 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
     4381 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     4441 gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac
     4501 agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc
     4561 atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg
     4621 ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat
     4681 gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat
     4741 cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt
     4801 gaattgctgc cctctggtta tgtgtgggag ggctaagcac ttcgtggccg aggagcagga
     4861 ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4921 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4981 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5041 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5101 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5161 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5221 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5281 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5341 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5401 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5461 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5521 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5581 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5641 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5701 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5761 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5821 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5881 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5941 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6001 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6061 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6121 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6181 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6241 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6301 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6361 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6421 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6481 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6541 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6601 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6661 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6721 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6781 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6841 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6901 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6961 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7021 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7081 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7141 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctca
//
