LOCUS       Exported                6407 bp DNA     circular SYN 30-AUG-2021
DEFINITION  A-to-G base editor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6407)
  TITLE     Liu Lab plasmids 20200102
REFERENCE   2  (bases 1 to 6407)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 30, 2021 from SnapGene 5.3.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6407
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        19..398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        399..602
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    635..644
                     /label=Kozak
     CDS             647..700
                     /codon_start=1
                     /label=BPNLS
                     /translation="KRTADGSEFESPKKKRKV"
     CDS             701..1198
                     /codon_start=1
                     /label=TadA
                     /translation="SEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEGWN
                     RPIGRHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSRIGRVVFG
                     ARDAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAALLSDFFRMRRQEIKAQKKAQS
                     STD"
     misc_feature    1199..1294
                     /label=Linker A
     CDS             1295..1792
                     /codon_start=1
                     /label=TadA* (8e)
                     /translation="SEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWN
                     RAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFG
                     VRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQS
                     SIN"
     misc_feature    1787..1789
                     /label=T166I
     misc_feature    1790..1792
                     /label=D167N
     misc_feature    1793..1888
                     /label=Linker B
     CDS             1889..1909
                     /label=SV40_NLS
     CDS             1910..1921
                     /label=GSGS linker
                     /label=GSG linker
     CDS             1922..3505
                     /codon_start=1
                     /label=dCasMINI-V4
                     /translation="AKNTITKTLKLRIVRPYNSAEVEKIVADEKNNREKIALEKNKDKV
                     KEACSKHLKVAAYCTTQVERNACLFCKARKLDDKFYQKLRGQFPDAVFWQEISEIFRQL
                     QKQAAEIYNQSLIELYYEIFIKGKGIANASSVEHYLSRVCYRRAAELFKNAAIASGLRS
                     KIKSNFRLKELKNMKSGLPTTKSDNFPIPLVKQKGGQYTGFEISNHNSDFIIKIPFGRW
                     QVKKEIDKYRPWEKFDFEQVQKSPKPISLLLSTQRRKRNKGWSKDEGTEAEIKKVMNGD
                     YQTSYIEVKRGSKICEKSAWMLNLSIDVPKIDKGVDPSIIGGIAVGVRSPLVCAINNAF
                     SRYSISDNDLFHFNKKMFARRRILLKKNRHKRAGHGAKNKLKPITILTEKSERFRKKLI
                     ERWACEIADFFIKNKVGTVQMENLESMKRKEDSYFNIRLRGFWPYAEMQNKIEFKLKQY
                     GIEIRKVAPNNTSKTCSKCGHLNNYFNFEYRKKNKFPHFKCEKCNFKENAAYNAALNIS
                     NPKLKSTKERP"
     misc_feature    2345..2347
                     /label=D143R
     misc_feature    2357..2359
                     /label=T147R
     modified_base   2894..2896
                     /label=D326A
                     /mod_base="OTHER"
     misc_feature    2906..2908
                     /label=K330R
     modified_base   3446..3448
                     /label=D510A
                     /mod_base="OTHER"
     misc_feature    3500..3502
                     /label=E528R
     misc_feature    3509..3535
                     /label=HA tag
     misc_feature    3551..3583
                     /label=linker
     CDS             3593..3607
                     /label=GS linker
     CDS             3614..3640
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-Myc NLS
     CDS             3641..3706
                     /label=3xFLAG
     polyA_signal    3735..3959
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4030..4046)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4054..4070
                     /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(4078..4108)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4123..4144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4432..5020)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5191..6051)
                     /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"
     promoter        complement(6052..6156)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
       61 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      121 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      181 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      241 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      301 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      361 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      421 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      481 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      541 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      601 ctggtttagt gaaccgtcag atccgctagc ccgggccacc atgggaaaac ggacagccga
      661 cggaagcgag ttcgagtcac caaagaagaa gcggaaagtc tctgaagtcg agtttagcca
      721 cgagtattgg atgaggcacg cactgaccct ggcaaagcga gcatgggatg aaagagaagt
      781 ccccgtgggc gccgtgctgg tgcacaacaa tagagtgatc ggagagggat ggaacaggcc
      841 aatcggccgc cacgacccta ccgcacacgc agagatcatg gcactgaggc agggaggcct
      901 ggtcatgcag aattaccgcc tgatcgatgc caccctgtat gtgacactgg agccatgcgt
      961 gatgtgcgca ggagcaatga tccacagcag gatcggaaga gtggtgttcg gagcacggga
     1021 cgccaagacc ggcgcagcag gctccctgat ggatgtgctg caccaccccg gcatgaacca
     1081 ccgggtggag atcacagagg gaatcctggc agacgagtgc gccgccctgc tgagcgattt
     1141 ctttagaatg cggagacagg agatcaaggc ccagaagaag gcacagagct ccaccgactc
     1201 tggaggatct agcggaggat cctctggaag cgagacacca ggcacaagcg agtccgccac
     1261 accagagagc tccggcggct cctccggagg atcctctgag gtggagtttt cccacgagta
     1321 ctggatgaga catgccctga ccctggccaa gagggcacgg gatgagaggg aggtgcctgt
     1381 gggagccgtg ctggtgctga acaatagagt gatcggcgag ggctggaaca gagccatcgg
     1441 cctgcacgac ccaacagccc atgccgaaat tatggccctg agacagggcg gcctggtcat
     1501 gcagaactac agactgattg acgccaccct gtacgtgaca ttcgagcctt gcgtgatgtg
     1561 cgccggcgcc atgatccact ctaggatcgg ccgcgtggtg tttggcgtga ggaactcaaa
     1621 aagaggcgcc gcaggctccc tgatgaacgt gctgaactac cccggcatga atcaccgcgt
     1681 cgaaattacc gagggaatcc tggcagatga atgtgccgcc ctgctgtgcg atttctatcg
     1741 gatgcctaga caggtgttca atgctcagaa gaaggcccag agctccatca actccggagg
     1801 atctagcgga ggatcctctg gcagcgagac accaggaaca agcgagtcag caacaccaga
     1861 gagcagtggc ggcagcagcg gcggcagccc caagaaaaaa cgcaaggtgg gaagcggatc
     1921 cgccaaaaac accattacca aaacactgaa actgcgtatt gtgcgtccgt ataatagcgc
     1981 agaagtggaa aaaattgttg ccgacgaaaa aaacaaccgc gaaaaaatcg cactggaaaa
     2041 gaacaaagac aaagtgaaag aagcctgcag caaacatctg aaagttgcag catattgtac
     2101 cacacaggtt gaacgtaatg catgcctgtt ttgtaaagca cgtaaactgg atgacaaatt
     2161 ctaccaaaaa ctgcgtggtc agtttccgga tgcagttttt tggcaagaaa tcagcgaaat
     2221 ttttcgccag ctgcagaaac aggcagcaga aatctataat cagagcctga tcgaactgta
     2281 ctacgagatt tttatcaaag gcaaaggtat tgcaaatgcc agcagcgttg aacattatct
     2341 gagtagagtt tgttatagac gtgcagcaga actgtttaaa aacgcagcaa ttgcaagcgg
     2401 tctgcgtagc aaaatcaaaa gcaattttcg tctgaaagaa ctgaaaaaca tgaaaagtgg
     2461 tctgccgacc accaaaagcg ataattttcc gattccgctg gttaaacaga aaggtggtca
     2521 gtataccggt tttgaaatta gcaatcataa tagcgacttc atcatcaaga ttccgtttgg
     2581 tcgttggcag gtcaaaaaag agattgataa atatcgtccg tgggagaaat ttgactttga
     2641 acaggttcag aaaagcccga aaccgattag cctgctgctg agcacccagc gtcgtaaacg
     2701 taataaaggt tggagcaaag atgaaggcac cgaagccgaa atcaaaaaag ttatgaatgg
     2761 cgattatcag accagctaca ttgaagttaa acgtggcagc aaaatctgtg aaaaaagcgc
     2821 atggatgctg aatctgagca ttgatgttcc gaaaattgat aaaggtgtgg atccgagcat
     2881 tattggtggt attgcagttg gtgttagatc accgctggtt tgcgcaatta acaatgcatt
     2941 tagccgttat agcatcagcg ataacgacct gtttcacttc aacaagaaaa tgtttgcacg
     3001 tcgtcgtatc ctgctgaaaa aaaaccgtca taaacgtgca ggtcatggtg caaaaaacaa
     3061 actgaaaccg atcaccattc tgaccgaaaa aagtgaacgt tttcgcaaaa agctgattga
     3121 acgttgggca tgtgaaatcg cggatttctt cattaaaaac aaagttggca ccgtgcagat
     3181 ggaaaatctg gaaagcatga aacgtaaaga ggacagctat tttaacattc gcctgcgtgg
     3241 cttttggccg tatgcagaaa tgcagaacaa aatcgaattc aaactgaagc agtatggcat
     3301 cgaaattcgt aaagttgcac cgaataatac cagcaaaacc tgtagcaaat gtggccatct
     3361 gaacaactat ttcaacttcg agtaccgcaa gaaaaacaaa ttcccgcact ttaaatgcga
     3421 aaaatgcaac ttcaaagaaa acgccgcgta taatgcagcc ctgaatattt caaacccgaa
     3481 actgaaaagc accaaagaga gaccggccta tccctatgac gtgcccgatt atgccagcct
     3541 gggcagcggc gacggcattg gtagtgggag caacggcagc agcctccata tgggtggagg
     3601 ctccggggaa gatcctgctg ccaaacgcgt taaactagac gactataagg accacgacgg
     3661 agactacaag gatcatgata ttgattacaa agacgatgac gataagtgag tttaaacccg
     3721 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3781 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3841 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3901 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3961 ttctgaggcg gaaagaacca gctggggctc gataccgtcg acctctagct agagcttggc
     4021 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4081 catacgagcc ggaagcataa agtgtaaagc ctagggtgcc taatgagtga gctaactcac
     4141 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4201 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4261 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4321 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4381 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4441 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4501 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4561 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4621 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4681 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4741 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4801 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4861 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4921 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4981 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5041 tacggggtct gacactcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5101 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5161 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5221 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5281 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5341 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5401 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5461 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5521 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5581 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5641 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5701 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5761 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5821 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5881 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5941 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6001 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6061 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6121 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6181 tgacgtcgac ggatcgggag atcgatctcc cgatccccta gggtcgactc tcagtacaat
     6241 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     6301 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     6361 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtac
//
