LOCUS       sequence-232620-        8292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent fusion protein used to visualise mouse BAK, with 
            hygromycin selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8292)
  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 8292)
  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-BAK
FEATURES             Location/Qualifiers
     source          1..8292
                     /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(2759..3333,3334..3336,3337..3345)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2926..2943)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3153..3172
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3371..4390
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             3371..4390
                     /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             4725..5239
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5401..5417
                     /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(5425..5431,5432..5449,5450..5455))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5470..5491
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5608..5625)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5779..6367)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5859..5878)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6538..7329,7330..7398))
                     /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     7161..7180
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7399..7503)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7571..7589
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7627..7649)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7749..7768
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7962..7984
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8112..8131
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8292,1..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 ggcatctgga caaggaccag gtcccccgaa ggtgggctgc gatgagtccc
     2161 cgtccccttc tgaacagcag gttgcccagg acacagagga ggtctttcga agctacgttt
     2221 tttacctcca ccagcaggaa caggagaccc agggggcggc cgcccctgcc aaccccgaga
     2281 tggacaactt gcccctggaa cccaacagca tcttgggtca ggtgggtcgg cagcttgctc
     2341 tcatcggaga tgatattaac cggcgctacg acacagagtt ccagaattta ctagaacagc
     2401 ttcagcccac agccgggaat gcctacgaac tcttcaccaa gatcgcctcc agcctattta
     2461 agagtggcat cagctggggc cgcgtggtgg ctctcctggg ctttggctac cgtctggccc
     2521 tgtacgtcta ccagcgtggt ttgaccggct tcctgggcca ggtgacctgc tttttggctg
     2581 atatcatact gcatcattac atcgccagat ggatcgcaca gagaggcggt tgggtggcag
     2641 ccctgaattt tcgtagagac cccatcctga ccgtaatggt gatttttggt gtggttctgt
     2701 tgggccaatt cgtggtacac agattcttca gatcatgact cgaggttaac gaattccgcc
     2761 cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     2821 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     2881 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     2941 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     3001 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     3061 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     3121 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     3181 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca
     3241 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     3301 acgtggtttt cctttgaaaa acacgatgat aagcttgcca caacccgtac caaagatgga
     3361 tagatccgga aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
     3421 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
     3481 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
     3541 agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga
     3601 cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
     3661 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat
     3721 ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
     3781 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
     3841 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
     3901 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
     3961 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
     4021 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
     4081 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
     4141 gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
     4201 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
     4261 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga
     4321 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
     4381 aaaggaatag agtagatgcc gaccgaacaa gagctgattt cgagaacgcc tcagccagca
     4441 actcgcgcga gcctagcaag gcaaatgcga gagaacggcc ttacgcttgg tggcacagtt
     4501 ctcgtccaca gttcgctaag ctcgctcggc tgggtcgcgg gagggccggt cgcagtgatt
     4561 caggcccttc tggattgtgt tggtccccag ggcacgattg tcatgcccac gcactcgggt
     4621 gatctgactg atcccgcaga ttggagatcg ccgcccgtgc ctgccgattg ggtgcagatc
     4681 cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     4741 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact
     4801 gagaatagag aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac
     4861 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag
     4921 atgcggtccc gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag
     4981 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
     5041 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc
     5101 agtcctccga tagactgcgt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt
     5161 gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc
     5221 gtcagcgggg gtctttcatg ggtaacagtt tcttgaagtt ggagaacaac attctgaggg
     5281 taggagtcga atattaagta atcctgactc aattagccac tgttttgaat ccacatactc
     5341 caatactcct gaaatagttc attatggaca gcgcagaaag agctggggag aattgtgaaa
     5401 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5461 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5521 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5581 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5641 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5701 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5761 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5821 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5881 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5941 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6001 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6061 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6121 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6181 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6241 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6301 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6361 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6421 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6481 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6541 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6601 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6661 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6721 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6781 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6841 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6901 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6961 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7021 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7081 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7141 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7201 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7261 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7321 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7381 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7441 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7501 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     7561 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     7621 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     7681 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     7741 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     7801 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     7861 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     7921 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     7981 aacgacggcg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     8041 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     8101 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     8161 atgccggcca cgatgcgtcc ggcgtagagg cgattagtcc aatttgttaa agacaggata
     8221 tcagtggtcc aggctctagt tttgactcaa caatatcacc agctgaagcc tatagagtac
     8281 gagccataga ta
//
