LOCUS       Exported                7850 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  expresses right-side TALE–DddAtox half ND4-DdCBE in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    ND4-DdCBE-right side TALE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7850)
  AUTHORS   Mok BY, de Moraes MH, Zeng J, Bosch DE, Kotrys AV, Raguram A, Hsu F,
            Radey MC, Peterson SB, Mootha VK, Mougous JD, Liu DR
  TITLE     A bacterial cytidine deaminase toxin enables CRISPR-free 
            mitochondrial base editing.
  JOURNAL   Nature. 2020 Jul 8. pii: 10.1038/s41586-020-2477-4. doi: 
            10.1038/s41586-020-2477-4.
  PUBMED    32641830
REFERENCE   2  (bases 1 to 7850)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7850
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             904..981
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MASVLTPLLLRGLTGSARRLPVPRAK"
     CDS             994..1059
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3634..3885
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     rep_origin      4082..4510
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4169..4188)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4379..4400
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4519..4539)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4524..4853
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4704..4839
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4766..4785
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4901..4948
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4967..5365
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5523..5644
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5560..5579)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5614..5633
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5693..5709)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5693..5709)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5706..5728)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5717..5733
                     /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(5741..5771)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5786..5807
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5924..5941)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6095..6683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6175..6194)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6854..7714)
                     /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     7477..7496
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7715..7819)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 accatggcct ccgttctgac cccgctgctg ctgcgtggcc tgaccggctc cgctcgtcgt
      961 ctgccagttc cgcgtgcgaa aatccattcc ctggactaca aagaccatga cggtgattat
     1021 aaagatcatg acatcgatta caaggatgac gatgacaaga tggacatcgc ggatctgcgt
     1081 accctgggtt acagccagca gcagcaggag aagatcaagc caaaggtgcg cagcaccgtg
     1141 gcccagcacc atgaagctct ggtgggtcac ggcttcaccc acgcgcacat cgtggctctg
     1201 agccagcacc cagccgcgct gggtaccgtg gccgtgaaat atcaggacat gattgctgcc
     1261 ctgccagagg ctacccatga agcgattgtg ggtgtgggca agcagtggag cggtgctcgt
     1321 gcgctggagg ccctgctgac cgtggccggt gaactgcgtg gcccgccgct gcagctggat
     1381 accggccagc tgctgaaaat cgcgaaacgt ggcggtgtga ccgctgtgga agctgtgcat
     1441 gcctggcgta atgctctgac cggtgccccg ctgaacctga ccccggaaca agtggttgcg
     1501 attgcgtccc atgatggtgg taaacaagcc ctggagaccg ttcaagctct gctgccagtg
     1561 ctgtgccagg ctcatggtct gaccccgcag caggtggtgg ctattgccag ccatgatggc
     1621 ggcaagcagg ctctggagac cgtgcagcgt ctgctgccgg tgctgtgcca agcccatggc
     1681 ctgaccccgc agcaagttgt ggctattgcc agcaacggcg gtggcaaaca ggcgctggaa
     1741 accgtgcagc gcctgttacc ggtgctgtgc caagcgcatg gtctgacccc ggagcaggtg
     1801 gtggccatcg ctagcaacaa cggtggcaag caggccctgg aaaccgtgca ggcgctgttg
     1861 ccggtgctgt gccaggccca tggcttaacc ccggaacaag tggtggccat tgcgagcaat
     1921 ggtggcggta aacaggctct ggaaaccgtg caggccctgc ttccggtgct gtgccaggcc
     1981 catgggctga ccccggaaca agttgtggct atcgccagca acatcggcgg caagcaggcg
     2041 ctggagaccg ttcaggctct gttaccggtg ctgtgccagg cgcatggcct gaccccggaa
     2101 caggtggtgg cgatcgctag caacattggc ggtaaacagg ccctggagac cgttcagcgc
     2161 ctgctcccgg tgctgtgcca ggcccatggt ctgaccccgg aacaggttgt tgctattgcc
     2221 agcaacaacg gcggcaagca ggccctggag accgtgcagg cgctgctacc ggtgctgtgc
     2281 caggcccatg gactgacccc ggagcaggtt gtggccatcg cgtccaatgg cggtggcaaa
     2341 caggctctgg agaccgttca gcgtctgtta ccggtgctgt gccaggcaca tggcctgacc
     2401 ccggagcaag ttgttgccat cgctagcaac attggtggca agcaggcgct ggaaaccgtt
     2461 cagcgcctgc taccggtgct gtgccaggct catggcttaa ccccggagca ggttgtcgcc
     2521 attgccagca acaatggtgg caaacaggct ctggaaactg tgcaggccct gctaccggtg
     2581 ctgtgccagg cccatgggtt aaccccggaa caggttgtgg ccattgcctc caataacggt
     2641 ggcaagcagg cgctggaaac ggtgcaggct ctgcttccgg tgctgtgcca ggctcatggg
     2701 ctgaccccgg agcaagtggt tgctattgcg tccaacattg gtggcaaaca ggccctggaa
     2761 accgttcagg cgctgctccc ggtgctgtgc caggcccatg ggctaacccc ggaacaggtg
     2821 gttgccattg cctccaacaa tggtggcaag caggccctgg aaacggttca gcgtctgctt
     2881 ccggtgctgt gccaggccca tgggcttacc ccgcagcaag ttgttgctat cgccagcaat
     2941 attggtggca aacaggctct ggaaacggtg cagcgcctgc taccggtgct gtgccaggct
     3001 catggtttaa ccccgcagca ggtggttgcg attgcctcca acaacggtgg caagcaggcg
     3061 ctggaaactg ttcagcgtct gctcccggtg ctgtgccagg ctcacggcct gaccccgcag
     3121 caagtggtgg ctatcgcctc caacggtggt ggtcgcccgg ctctggaaag cattgtggcg
     3181 cagctgagcc gtccagaccc ggccctggcg gctctgacca atgatcacct ggtggccctg
     3241 gcctgcctgg gtggccgtcc ggctctggat gccgtgaaga aaggtctggg cggatccggc
     3301 agctacgccc tgggtccgta tcagattagc gccccgcagc tgccagctta caatggtcag
     3361 accgtgggta ccttctacta tgtgaacgac gcgggcggtc tggagagcaa ggtgtttagc
     3421 agcggcggtc caaccccgta cccaaactat gccaatgccg gtcatgtgga gggtcagagc
     3481 gccctgttca tgcgtgataa cggcatcagc gagggtctgg tgttccacaa caacccggaa
     3541 ggcacctgcg gtttttgcgt gaacatgacc gagaccctgc tgccggaaaa cgcgaaaatg
     3601 accgtggtgc cgccggaagg ttctggcggc tcaactaatc tgagcgacat cattgagaag
     3661 gagactggga aacagctggt cattcaggag tccatcctga tgctgcctga ggaggtggag
     3721 gaagtgatcg gcaacaagcc agagtctgac atcctggtgc acaccgccta cgacgagtcc
     3781 acagatgaga atgtgatgct gctgacctct gacgcccccg agtataagcc ttgggccctg
     3841 gtcatccagg attctaacgg cgagaataag atcaagatgc tgtgataagg ggtctttgtc
     3901 ctctgtactg tctctctcct tgcccctaac ccaaaaagct tcatttttct gtgtaggctg
     3961 cacaagagcc ttgattgaag atatattctt tctgaacagt atttaaggtt tccaataaaa
     4021 tgtacacccc tcagaacttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     4081 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4141 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4201 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4261 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4321 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4381 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4441 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4501 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4561 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4621 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4681 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4741 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4801 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4861 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
     4921 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagccttt
     4981 gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat
     5041 ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg
     5101 tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc
     5161 tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg
     5221 catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa
     5281 agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc
     5341 ctctggttat gtgtgggagg gctaagcact tcgtggccga ggagcaggac tgacacgtgc
     5401 tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5461 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5521 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5581 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5641 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5701 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5761 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5821 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5881 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5941 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6001 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6061 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6121 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6181 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6241 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6301 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6361 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6421 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6481 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6541 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6601 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6661 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6721 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6781 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6841 cttggtctga cagttaccaa tgtttaatca gtgaggcacc tatctcagcg atctgtctat
     6901 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6961 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7021 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7081 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7141 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7201 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7261 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7321 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7381 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7441 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7501 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7561 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7621 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7681 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7741 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7801 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
