LOCUS       sequence-226691-        8340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fusion protein used to visualise mitochondria upon the addition of 
            Halo specific dye, has hygromycin selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8340)
  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 8340)
  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-TOMM20-Halo
FEATURES             Location/Qualifiers
     source          1..8340
                     /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"
     regulatory      1416..1425
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1890..2780
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     misc_feature    join(2809..3383,3384..3386,3387..3395)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2976..2993)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3203..3222
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             4774..5288
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5449..5465
                     /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(5473..5479,5480..5497,5498..5503))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5518..5539
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5656..5673)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5827..6415)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5907..5926)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6586..7377,7378..7446))
                     /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     7209..7228
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7447..7551)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7619..7637
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7675..7697)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7797..7816
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8160..8179
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8340,1..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 catggtgggt cggaacagcg
     1441 ccatcgccgc cggtgtatgc ggggcccttt tcattgggta ctgcatctac ttcgaccgca
     1501 aaagacgaag tgaccccaac ttcaagaaca ggcttcgaga acgaagaaag aaacagaagc
     1561 ttgccaagga gagagctggg ctttccaagt tacctgacct taaagatgct gaagctgttc
     1621 agaagttctt ccttgaagaa atacagcttg gtgaagagtt actagctcaa ggtgaatatg
     1681 agaagggcgt agaccatctg acaaatgcaa ttgctgtgtg tggacagcca cagcagttac
     1741 tgcaggtctt acagcaaact cttccaccac cagtgttcca gatgcttctg actaagctcc
     1801 caacaattag tcagagaatt gtaagtgctc agagcttggc tgaagatgat gtggaagtcg
     1861 acggtaccgc gggcccggga tccaccggta tggcagaaat cggtactggc tttccattcg
     1921 acccccatta tgtggaagtc ctgggcgagc gcatgcacta cgtcgatgtt ggtccgcgcg
     1981 atggcacccc tgtgctgttc ctgcacggta acccgacctc ctcctacgtg tggcgcaaca
     2041 tcatcccgca tgttgcaccg acccatcgct gcattgctcc agacctgatc ggtatgggca
     2101 aatccgacaa accagacctg ggttatttct tcgacgacca cgtccgcttc atggatgcct
     2161 tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat tcacgactgg ggctccgctc
     2221 tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa aggtattgca tttatggagt
     2281 tcatccgccc tatcccgacc tgggacgaat ggccagaatt tgcccgcgag accttccagg
     2341 ccttccgcac caccgacgtc ggccgcaagc tgatcatcga tcagaacgtt tttatcgagg
     2401 gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt cgagatggac cattaccgcg
     2461 agccgttcct gaatcctgtt gaccgcgagc cactgtggcg cttcccaaac gagctgccaa
     2521 tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga atacatggac tggctgcacc
     2581 agtcccctgt cccgaagctg ctgttctggg gcaccccagg cgttctgatc ccaccggccg
     2641 aagccgctcg cctggccaaa agcctgccta actgcaaggc tgtggacatc ggcccgggtc
     2701 tgaatctgct gcaagaagac aacccggacc tgatcggcag cgagatcgcg cgctggctgt
     2761 cgacgcttga aatttccggc tgactcgagt cgaggttaac gaattccgcc cctctccctc
     2821 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     2881 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     2941 tgttttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     3001 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     3061 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     3121 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     3181 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     3241 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
     3301 atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     3361 cctttgaaaa acacgatgat aagcttgcca caacccgtac caaagatgga tagatccgga
     3421 aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc
     3481 tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga
     3541 gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat
     3601 gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa
     3661 ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac
     3721 ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc
     3781 gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt
     3841 caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg
     3901 caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc agctctcgat gagctgatgc
     3961 tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca
     4021 atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg
     4081 gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg
     4141 agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc
     4201 gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt
     4261 tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga
     4321 ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag
     4381 aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaataga
     4441 gtagatgccg accgaacaag agctgatttc gagaacgcct cagccagcaa ctcgcgcgag
     4501 cctagcaagg caaatgcgag agaacggcct tacgcttggt ggcacagttc tcgtccacag
     4561 ttcgctaagc tcgctcggct gggtcgcggg agggccggtc gcagtgattc aggcccttct
     4621 ggattgtgtt ggtccccagg gcacgattgt catgcccacg cactcgggtg atctgactga
     4681 tcccgcagat tggagatcgc cgcccgtgcc tgccgattgg gtgcagatcc gataaaataa
     4741 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     4801 aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg agaatagaga
     4861 agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca ggatatctgt
     4921 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtcccg
     4981 ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg
     5041 accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc
     5101 tgctccccga gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat
     5161 agactgcgtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt
     5221 gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg
     5281 tctttcatgg gtaacagttt cttgaagttg gagaacaaca ttctgagggt aggagtcgaa
     5341 tattaagtaa tcctgactca attagccact gttttgaatc cacatactcc aatactcctg
     5401 aaatagttca ttatggacag cgcagaagag ctggggagaa ttgtgaaatt gttatccgct
     5461 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5521 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5581 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5641 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5701 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5761 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5821 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5881 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5941 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6001 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6061 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6121 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6181 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6241 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6301 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6361 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6421 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6481 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6541 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6601 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6661 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6721 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6781 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6841 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6901 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6961 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7021 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7081 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7141 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7201 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7261 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7321 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7381 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7441 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7501 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7561 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     7621 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     7681 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     7741 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     7801 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     7861 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     7921 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     7981 gcgattaagt tgggtaacgc cagggttttt aggttgaggc cgttgagcac cgccgccgca
     8041 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     8101 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     8161 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
     8221 atgcgtccgg cgtagaggcg attagtccaa tttgttaaag acaggatatc agtggtccag
     8281 gctctagttt tgactcaaca atatcaccag ctgaagccta tagagtacga gccatagata
//
