LOCUS       sequence-226689-        8322 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 8322)
  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 8322)
  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..8322
                     /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"
     LTR             4724..5238
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(5407..5423)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5407..5423)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5420..5442)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5431..5447
                     /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(5455..5461,5462..5479,5480..5485))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5500..5521
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5638..5655)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5809..6397)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5889..5908)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6568..7359,7360..7428))
                     /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     7191..7210
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7429..7533)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7601..7619
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7657..7679)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7779..7798
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7992..8014
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8142..8161
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8322,1..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 tgttttcttg 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 agctctcgat
     3901 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc
     3961 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag
     4021 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg
     4081 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg
     4141 ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt
     4201 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc
     4261 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat
     4321 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca
     4381 aaggaataga gtagatgccg accgaacaag agctgatttc gagaacgcct cagccagcaa
     4441 ctcgcgcgag cctagcaagg caaatgcgag agaacggcct tacgcttggt ggcacagttc
     4501 tcgtccacag ttcgctaagc tcgctcggct gggtcgcggg agggccggtc gcagtgattc
     4561 aggcccttct ggattgtgtt ggtccccagg gcacgattgt catgcccacg cactcgggtg
     4621 atctgactga tcccgcagat tggagatcgc cgcccgtgcc tgccgattgg gtgcagatcc
     4681 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg
     4741 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg
     4801 agaatagaga agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca
     4861 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
     4921 tgcggtcccg ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
     4981 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt
     5041 cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact cggcgcgcca
     5101 gtcctccgat agactgcgtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg
     5161 catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg
     5221 tcagcggggg tctttcatgg gtaacagttt cttgaagttg gagaacaaca ttctgagggt
     5281 aggagtcgaa tattaagtaa tcctgactca attagccact gttttgaatc cacatactcc
     5341 aatactcctg aaatagttca ttatggacag cgcagaagag ctggggagaa ttaattcgta
     5401 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5461 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5521 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5581 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5641 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5701 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5761 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5821 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5881 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5941 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6001 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6061 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6121 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6181 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6241 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6301 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6361 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6421 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6481 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6541 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6601 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6661 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6721 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6781 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6841 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6901 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6961 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7021 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7081 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7141 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7201 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7261 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7321 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7381 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7441 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7501 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7561 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     7621 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     7681 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     7741 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     7801 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     7861 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     7921 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     7981 gccagggttt tcccagtcac gacgttgtaa aacgacggcg caaggaatgg tgcatgcaag
     8041 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     8101 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     8161 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     8221 cgattagtcc aatttgttaa agacaggata tcagtggtcc aggctctagt tttgactcaa
     8281 caatatcacc agctgaagcc tatagagtac gagccataga ta
//
