LOCUS       sequence-232197-        8241 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent fusion protein used to visualise mouse BAX, with 
            hygromycin selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8241)
  AUTHORS   McArthur K, Whitehead LW, Heddleston JM, Li L, Padman BS, Oorschot 
            V, Geoghegan ND, Chappaz S, Davidson S, San Chin H, Lane RM, 
            Dramicanin M, Saunders TL, Sugiana C, Lessene R, Osellame LD, Chew 
            TL, Dewson G, Lazarou M, Ramm G, Lessene G, Ryan MT, Rogers KL, van 
            Delft MF, Kile BT
  TITLE     BAK/BAX macropores facilitate mitochondrial herniation and mtDNA 
            efflux during apoptosis.
  JOURNAL   Science. 2018 Feb 23;359(6378). pii: 359/6378/eaao6047. doi: 
            10.1126/science.aao6047.
  PUBMED    29472455
REFERENCE   2  (bases 1 to 8241)
  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: "Science."; 
            date: "2018-02-23"; volume: "359(6378). pii: 359/6378/eaao6047. 
            doi"; pages: " 10.1126/science.aao6047"
COMMENT     Sequence Label: pMIH-mRuby2-BAX
FEATURES             Location/Qualifiers
     source          1..8241
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(2708..3282,3283..3285,3286..3294)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2875..2892)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3102..3121
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3320..4339
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             3320..4339
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     LTR             4674..5188
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5350..5366
                     /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(5374..5380,5381..5398,5399..5404))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5419..5440
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5557..5574)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5728..6316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5808..5827)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6487..7278,7279..7347))
                     /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     7110..7129
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7348..7452)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7520..7538
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7576..7598)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7698..7717
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7911..7933
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8061..8080
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8241,1..514)
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatctgccac catgaacagc ctgatcaagg
     1441 aaaatatgcg tatgaaggtg gtcatggaag gttcggtcaa cggccaccaa ttcaaatgca
     1501 caggtgaagg agaaggcaat ccgtacatgg gaactcaaac catgaggatc aaagtcatcg
     1561 agggaggacc cctgccattt gcctttgaca ttcttgccac gtcgttcatg tatggcagcc
     1621 gtacttttat caagtacccg aaaggcattc ctgatttctt taaacagtcc tttcctgagg
     1681 gttttacttg ggaaagagtt acgagatacg aagatggtgg agtcgtcacc gtcatgcagg
     1741 acaccagcct tgaggatggc tgtctcgttt accacgtcca agtcagaggg gtaaactttc
     1801 cctccaatgg tcccgtgatg cagaagaaga ccaagggttg ggagcctaat acagagatga
     1861 tgtatccagc agatggtggt ctgaggggat acactcatat ggcactgaaa gttgatggtg
     1921 gtggccatct gtcttgctct ttcgtaacaa cttacaggtc aaaaaagacc gtcgggaaca
     1981 tcaagatgcc cggtatccat gccgttgatc accgcctgga aaggttagag gaaagtgaca
     2041 atgaaatgtt cgtagtacaa cgcgaacacg cagttgccaa gttcgccggg cttggtggtg
     2101 ggaccggtat ggacgggtcc ggggagcagc ttgggagcgg cgggcccacc agctctgaac
     2161 agatcatgaa gacaggggcc tttttgctac agggtttcat ccaggatcga gcagggagga
     2221 tggctgggga gacacctgag ctgaccttgg agcagccgcc ccaggatgcg tccaccaaga
     2281 agctgagcga gtgtctccgg cgaattggag atgaactgga cagcaatatg gagctgcaga
     2341 ggatgattgc tgacgtggac acggactccc cccgagaggt cttcttccgg gtggcagctg
     2401 acatgtttgc tgatggcaac ttcaactggg gccgcgtggt tgccctcttc tactttgcta
     2461 gcaaactggt gctcaaggcc ctgtgcacta aagtgcccga gctgatcaga accatcatgg
     2521 gctggacact ggacttcctc cgtgagcggc tgcttgtctg gatccaagac cagggtggct
     2581 gggaaggcct cctctcctac ttcgggaccc ccacatggca gacagtgacc atctttgtgg
     2641 ctggagtcct caccgcctcg ctcaccatct ggaagaagat gggctgactc gaggttaacg
     2701 aattccsccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     2761 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     2821 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     2881 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     2941 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     3001 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     3061 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     3121 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     3181 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     3241 accacgggga cgtggttttc ctttgaaaaa cacgatgata agcttgccac aacccgtacc
     3301 aaagatggat agatccggaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     3361 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     3421 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     3481 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
     3541 agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     3601 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     3661 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
     3721 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     3781 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     3841 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     3901 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     3961 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     4021 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     4081 tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     4141 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     4201 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt
     4261 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     4321 tccgagggca aaggaataga gtagatgccg accgaacaag agctgatttc gagaacgcct
     4381 cagccagcaa ctcgcgcgag cctagcaagg caaatgcgag agaacggcct tacgcttggt
     4441 ggcacagttc tcgtccacag ttcgctaagc tcgctcggct gggtcgcggg agggccggtc
     4501 gcagtgattc aggcccttct ggattgtgtt ggtccccagg gcacgattgt catgcccacg
     4561 cactcgggtg atctgactga tcccgcagat tggagatcgc cgcccgtgcc tgccgattgg
     4621 gtgcagatcc gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag
     4681 accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa
     4741 tacataactg agaatagaga agttcagatc aaggttagga acagagagac agcagaatat
     4801 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     4861 ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat gtttccaggg
     4921 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
     4981 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
     5041 cggcgcgcca gtcctccgat agactgcgtc gcccgggtac ccgtgtatcc aataaaccct
     5101 cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
     5161 tgactacccg tcagcggggg tctttcatgg gtaacagttt cttgaagttg gagaacaaca
     5221 ttctgagggt aggagtcgaa tattaagtaa tcctgactca attagccact gttttgaatc
     5281 cacatactcc aatactcctg aaatagttca ttatggacag cgcagaaaga gctggggaga
     5341 attgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5401 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5461 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5521 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5581 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5641 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5701 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5761 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5821 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5881 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5941 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6001 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6061 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6121 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6181 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6241 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6301 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6361 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6421 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6481 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6541 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6601 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6661 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6721 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6781 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6841 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6901 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6961 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7021 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7081 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7141 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7201 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7261 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7321 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7381 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7441 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     7501 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     7561 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     7621 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     7681 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     7741 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     7801 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     7861 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     7921 acgttgtaaa acgacggcgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     7981 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     8041 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     8101 gcgccggtga tgccggccac gatgcgtccg gcgtagaggc gattagtcca atttgttaaa
     8161 gacaggatat cagtggtcca ggctctagtt ttgactcaac aatatcacca gctgaagcct
     8221 atagagtacg agccatagat a
//
