LOCUS       sequence-226688-        8269 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 8269)
  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 8269)
  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..8269
                     /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(2705..3279,3280..3282,3283..3291)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2872..2889)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3099..3118
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             4670..5185
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(5354..5370)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5354..5370)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5367..5389)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5378..5394
                     /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(5402..5408,5409..5426,5427..5432))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5447..5468
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5585..5602)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5756..6344)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5836..5855)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6515..7306,7307..7375))
                     /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     7138..7157
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7376..7480)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7548..7566
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7604..7626)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7726..7745
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7939..7961
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8089..8108
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8269,1..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 gggcctcgag gttaacgaat
     2701 tccgcccctc tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc
     2761 ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg
     2821 cccggaaacc tggccctgtt ttcttgacga gcattcctag gggtctttcc cctctcgcca
     2881 aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa
     2941 gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt
     3001 gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt
     3061 gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca
     3121 acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc
     3181 ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc
     3241 acggggacgt ggttttcctt tgaaaaacac gatgataagc ttgccacaac ccgtaccaaa
     3301 gatggataga tccggaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga
     3361 aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt
     3421 cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt
     3481 ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt
     3541 gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg
     3601 tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga
     3661 ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg
     3721 accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc
     3781 ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcagct
     3841 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
     3901 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
     3961 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
     4021 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
     4081 ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
     4141 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
     4201 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg
     4261 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
     4321 agggcaaagg aatagagtag atgccgaccg aacaagagct gatttcgaga acgcctcagc
     4381 cagcaactcg cgcgagccta gcaaggcaaa tgcgagagaa cggccttacg cttggtggca
     4441 cagttctcgt ccacagttcg ctaagctcgc tcggctgggt cgcgggaggg ccggtcgcag
     4501 tgattcaggc ccttctggat tgtgttggtc cccagggcac gattgtcatg cccacgcact
     4561 cgggtgatct gactgatccc gcagattgga gatcgccgcc cgtgcctgcc gattgggtgc
     4621 agatccgata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc
     4681 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaataca
     4741 taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc
     4801 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc
     4861 cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc
     4921 ccaaggacct gaaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     4981 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     5041 cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt
     5101 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     5161 ctacccgtca gcgggggtct ttcatgggta acagtttctt gaagttggag aacaacattc
     5221 tgagggtagg agtcgaatat taagtaatcc tgactcaatt agccactgtt ttgaatccac
     5281 atactccaat actcctgaaa tagttcatta tggacagcgc agaagagctg gggagaatta
     5341 attcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5401 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5461 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5521 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5581 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5641 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5701 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5761 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5821 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5881 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5941 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6001 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6061 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6121 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6181 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6241 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6301 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6361 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6421 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6481 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6541 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6601 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6661 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6721 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6781 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6841 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6901 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6961 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7021 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7081 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7141 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7201 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7261 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7321 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7381 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7441 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7501 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     7561 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     7621 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     7681 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     7741 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     7801 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     7861 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     7921 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggcgcaa ggaatggtgc
     7981 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     8041 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     8101 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     8161 gtagaggcga ttagtccaat ttgttaaaga caggatatca gtggtccagg ctctagtttt
     8221 gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagata
//
