LOCUS       sequence-170396-        5274 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of ATP8 with mitochondrial targeting sequence, 
            Myc, and FLAG tags.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5274)
  AUTHORS   Boominathan A, Vanhoozer S, Basisty N, Powers K, Crampton AL, Wang 
            X, Friedricks N, Schilling B, Brand MD, O'Connor MS
  TITLE     Stable nuclear expression of ATP8 and ATP6 genes rescues a mtDNA 
            Complex V null mutant.
  JOURNAL   Nucleic Acids Res. 2016 Nov 2;44(19):9342-9357. Epub 2016 Sep 4.
  PUBMED    27596602
REFERENCE   2  (bases 1 to 5274)
  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: "Nucleic 
            Acids Res."; date: "2016-11-2"; volume: "44(19)"; pages: "9342-9357.
            Epub 2016 Sep 4"
COMMENT     Sequence Label: pCMV6-G1-ATP8-Myc-FLAG (G418)
FEATURES             Location/Qualifiers
     source          1..5274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..173
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     165..182
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     877..897
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     952..971
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1440..1469
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1488..1511
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1531..1547)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1531..1547)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1558..2180
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1757..1776)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(2329..2917)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2409..2428)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3246..3293
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3318..3337
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3525..4319)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(3525..4319)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3636..3655)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4246..4265
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4354..4711)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4368..4503
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4520..4711
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4668..4688
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4713..4817)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4844..5272
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4931..4950)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5141..5162
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg
     1021 cgatcgccat gcagaccgcc ggggcattat tcatttctcc agctctgatc cgctgttgta
     1081 ccaggggtct aatcaggcct gtgtctgcct ccttcttgaa tagcccagtg aattcatcta
     1141 aacagccttc ctacagcaac ttcccactcc aggtggccag acgggagttc cagaccagtg
     1201 ttgtctcccg ggacattgac acggcgacgc gtatgcccca gctgaacacc acagtgtggc
     1261 ctactatgat caccccaatg ctgctgacac tgttcctgat tactcagctg aagatgctga
     1321 acaccaatta ccacctgccc ccttccccaa aacccatgaa gatgaaaaac tataataagc
     1381 cctgggaacc taagtggacc aaaatctgta gcctgcatag cctgccccct cagtcactcg
     1441 agcagaaact catctcagaa gaggatctgg cagcaaatga tatcctggat tacaaggatg
     1501 acgacgataa ggtttaaacg gccggccgcg gtcatagctg tttcctgaac agatcccggg
     1561 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     1621 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     1681 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     1741 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     1801 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
     1861 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     1921 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     1981 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
     2041 ttaaaataac tataccagca ggaggacgtc cagacacagc ataggctacc tggccatgcc
     2101 caaccggtgg gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact
     2161 cagtagatgc ctgttgaatt gggtacgcgg ccagcggcga gcggtatcag ctcactcaaa
     2221 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2281 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2341 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2401 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2461 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2521 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2581 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2641 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2701 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2761 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2821 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2881 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2941 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3001 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3061 tatatatgag taacctgagg ctatggcagg gcctgccgcc ccgacgttgg ctgcgagccc
     3121 tgggccttca cccgaacttg gggggtgggg tggggaaaag gaagaaacgc gggcgtattg
     3181 gccccaatgg ggtctcggtg gggtatcgac agagtgccag ccctgggacc gaaccccgcg
     3241 tttatgaaca aacgacccaa caccgtgcgt tttattctgt ctttttattg ccgtcatagc
     3301 gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc tagggtgggc
     3361 gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac
     3421 gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag
     3481 gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca
     3541 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
     3601 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
     3661 tcctgatagc gatccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
     3721 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
     3781 tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
     3841 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
     3901 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
     3961 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     4021 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     4081 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc
     4141 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
     4201 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
     4261 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg
     4321 cgaaacgatc ctcatcctgt ctcttgatcg atctttgcaa aagcctaggc ctccaaaaaa
     4381 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat
     4441 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
     4501 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
     4561 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
     4621 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
     4681 cacattccac agctggttct ttccgcctca ggactcttcc tttttcaata ttattgaagc
     4741 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4801 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc
     4861 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4921 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4981 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     5041 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5101 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5161 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     5221 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa tttt
//
