LOCUS       sequence-193041-        6077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mitochondria  targetted Nix .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6077)
  AUTHORS   Mughal W, Martens M, Field J, Chapman D, Huang J, Rattan S, Hai Y, 
            Cheung KG, Kereliuk S, West AR, Cole LK, Hatch GM, Diehl-Jones W, 
            Keijzer R, Dolinsky VW, Dixon IM, Parmacek MS, Gordon JW
  TITLE     Myocardin regulates mitochondrial calcium homeostasis and prevents 
            permeability transition.
  JOURNAL   Cell Death Differ. 2018 Mar 6. pii: 10.1038/s41418-018-0073-z. doi: 
            10.1038/s41418-018-0073-z.
  PUBMED    29511336
REFERENCE   2  (bases 1 to 6077)
  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: "Cell Death 
            Differ. 2018 Mar 6. pii: 10.1038/s41418-018-0073-z. doi: 
            10.1038/s41418-018-0073-z."
COMMENT     Sequence Label: Myc-Nix-MaoB
FEATURES             Location/Qualifiers
     source          1..6077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..406
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        407..610
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     561..581
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     655..674
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        655..673
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             696..725
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(1422..1440)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1423..1440)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1460..1477)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1466..1690
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1736..2164
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1823..1842)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2033..2054
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2173..2193)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2178..2507
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2358..2493
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2420..2439
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2574..3368
                     /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(2628..2647)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3238..3257
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3542..3663
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3579..3598)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3633..3652
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3712..3728)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3712..3728)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3725..3747)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3736..3752
                     /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(3760..3766,3767..3784,3785..3790))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3805..3826
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3943..3960)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4114..4702)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4194..4213)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4873..5664,5665..5733))
                     /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     5496..5515
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5734..5838)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5913..5932)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
       61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      361 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      421 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      481 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      541 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      601 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
      661 gactcactat agggagaccc aagcttggta ccatggagca gaagctgatc tccgaggagg
      721 acctgggatc catcgatgtc gacgaattca tgtcgtccca cctagtcgag ccgccgccgc
      781 ccctgcacaa caacaacaac aactgcgagg aaaatgagca gtctctgccc ccgccggccg
      841 gcctcaacag ttcctgggtg gagctaccca tgaacagcag caatggcaat gataatggca
      901 atgggaaaaa tggggggctg gaacacgtac catcctcatc ctccatccac aatggagaca
      961 tggagaagat tcttttggat gcacaacatg aatcaggaca gagtagttcc agaggcagtt
     1021 ctcactgtga cagcccttcg ccacaagaag atgggcagat catgtttgat gtggaaatgc
     1081 acaccagcag ggaccatagc tctcagtcag aagaagaagt tgtagaagga gagaaggaag
     1141 tcgaggcttt gaagaaaagt gcggactggg tatcagactg gtccagtaga cccgaaaaca
     1201 ttccacccaa ggagttccac ttcagacacc ctaaacgttc tgtgtcttta agcatgagga
     1261 aaagtggagc catgaagaaa gggggtattt tctccgcaga atttctgctc aggctgattg
     1321 gattgaccac catcttttca gcaacggctc ttggcttcct ggcccacaaa agggggctac
     1381 ttgtgagagt ctaactcgag catgcatcta gagggcccta ttctatagtg tcacctaaat
     1441 gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     1501 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     1561 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     1621 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     1681 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     1741 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1801 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1861 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1921 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     1981 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2041 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     2101 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2161 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2221 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     2281 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     2341 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2401 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     2461 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     2521 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     2581 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     2641 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     2701 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     2761 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     2821 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     2881 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     2941 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     3001 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     3061 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     3121 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     3181 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     3241 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     3301 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     3361 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     3421 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     3481 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     3541 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3601 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3661 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     3721 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3781 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3841 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3901 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3961 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4021 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4081 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4141 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4201 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4261 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4321 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4381 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4441 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4501 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4561 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4621 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4681 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4741 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4801 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4861 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4921 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4981 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5041 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5101 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5161 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5221 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5281 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5341 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5401 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5461 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5521 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5581 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5641 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5701 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5761 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5821 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     5881 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     5941 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     6001 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
     6061 cgttttgcgc tgcttcg
//
