LOCUS       sequence-356294-        6411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mitoZFD_ND2-Right-G1397-C(DddAtox half) in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6411)
  AUTHORS   Lim K, Cho SI, Kim JS
  TITLE     Nuclear and mitochondrial DNA editing in human cells with zinc 
            finger deaminases.
  JOURNAL   Nat Commun. 2022 Jan 18;13(1):366. doi: 10.1038/s41467-022-27962-0.
  PUBMED    35042880
REFERENCE   2  (bases 1 to 6411)
  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 
            Commun."; date: "2022-01-18"; volume: "13(1):366. doi"; pages: " 
            10.1038/s41467-022-27962-0"
COMMENT     Sequence Label: mitoZFD_ND2-Right-G1397-C
FEATURES             Location/Qualifiers
     source          1..6411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             1048..1074
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             1048..1074
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1678..1929
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     promoter        complement(1960..1978)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1961..1978)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1998..2015)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2004..2228
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2274..2702
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2361..2380)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2571..2592
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2711..2731)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2716..3045
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2896..3031
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2958..2977
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3116..3910
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3170..3189)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3780..3799
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4084..4205
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4121..4140)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4175..4194
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4254..4270)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4254..4270)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4267..4289)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4278..4294
                     /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(4302..4308,4309..4326,4327..4332))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4347..4368
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4485..4502)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4656..5244)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4736..4755)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5415..6206,6207..6275))
                     /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     6038..6057
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6276..6380)
                     /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 gcttgccacc
      901 atgttggggt ttgtgggtcg ggtggccgct gctccggcct ccggggcctt gcggagactc
      961 accccttcag cgtcgctgcc cccagctcag ctcttactgc gggccgctcc gacggcggtc
     1021 catcctgtca gggactatgc ggcgcaatac ccctacgacg tgcccgacta cgccgtggat
     1081 gaaatgacca aaaagttcgg cacgctcacc attcacgaca ccgaaaaggc cgccgaattc
     1141 ggcattcatg gggtacccgc cgctatggga gccataccgg ttaagcgcgg agcgactggt
     1201 gagaccaagg tgttcacggg gaactccaat tcccccaaga gccccactaa aggaggctgc
     1261 tccgggacac cacacgaagt aggagtttac accctgtctg gcacacctca cgaggtggga
     1321 gtgtacaccc tgtacgagtg cgaccactgt ggcaagtcct tcagccagtc tagccacctg
     1381 aacgtgcata agagaaccca cacaggggaa aagccttacc actgtgattg ggacggctgc
     1441 ggctggaagt tcgccagaag cgacgagctg accagacact accggaagca cacaggcgag
     1501 aaaccttaca aatgccccga ttgcggcaag tcattttctc agtcatctag cctgatcaga
     1561 caccagcgga cccacacggg cgagaagccc tacaagtgcg gccagtgtgg caagttctac
     1621 agccaggtgt ctcacctgac cagacaccag aaaatccacc tgcggtccgg tggatccaca
     1681 aatctgtctg acatcataga aaaagagacc ggcaaacaac tggtgataca agaaagtatt
     1741 ctgatgttgc ctgaggaagt tgaagaggta ataggcaaca agcctgaaag cgatatactg
     1801 gtacataccg catacgatga gtcaactgac gaaaacgtga tgctcttgac atccgatgcc
     1861 cctgaataca agccctgggc acttgtaatc caagatagta atggggagaa caaaatcaaa
     1921 atgttgtaat agctcgagca tgcatctaga gggccctatt ctatagtgtc acctaaatgc
     1981 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2041 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2101 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2161 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     2221 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     2281 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2341 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2401 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2461 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2521 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2581 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2641 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2701 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2761 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2821 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2881 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2941 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3001 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccggccggg
     3061 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     3121 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3181 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3241 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     3301 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3361 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3421 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3481 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3541 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     3601 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     3661 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     3721 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     3781 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     3841 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     3901 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     3961 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4021 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4081 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4141 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4201 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     4261 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4321 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4381 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4441 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4501 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4561 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4621 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4681 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4741 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4801 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4861 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4921 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4981 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5041 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5101 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5161 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5221 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5281 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5341 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5401 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5461 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5521 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5581 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5641 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5701 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5761 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5821 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5881 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5941 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6001 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6061 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6121 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6181 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6241 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6301 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6361 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
