LOCUS       sequence-399768-        6455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retrovirus construct for stable expressing mt-mKeima.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6455)
  AUTHORS   Lazarou M, Sliter DA, Kane LA, Sarraf SA, Wang C, Burman JL, Sideris
            DP, Fogel AI, Youle RJ
  TITLE     The ubiquitin kinase PINK1 recruits autophagy receptors to induce 
            mitophagy.
  JOURNAL   Nature. 2015 Aug 20;524(7565):309-14. doi: 10.1038/nature14893. Epub
            2015 Aug 12.
  PUBMED    26266977
REFERENCE   2  (bases 1 to 6455)
  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"; doi: 
            "10.1038/nature14893"; journalName: "Nature"; date: "2015-08-20- 
            20"; volume: "524"; issue: "7565"; pages: "309-14"
COMMENT     Sequence Label: pCHAC-mt-mKeima
FEATURES             Location/Qualifiers
     source          1..6455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             514..983
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    1048..1405
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1470..1886
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1816..1838
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1854..1870
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1896..2270
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     2208..2224
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             2303..2377
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             2411..2485
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             join(2528..2530,2531..2533,2534..3196)
                     /codon_start=1
                     /product="humanized monomeric Keima-Red fluorescent 
                     protein"
                     /label=hmKeima-Red
                     /note="mammalian codon-optimized"
                     /translation="MVSVIAKQMTYKVYMSGTVNGHYFEVEGDGKGKPYEGEQTVKLTV
                     TKGGPLPFAWDILSPQLQYGSIPFTKYPEDIPDYFKQSFPEGYTWERSMNFEDGAVCTV
                     SNDSSIQGNCFIYNVKISGENFPPNGPVMQKKTQGWEPSTERLFARDGMLIGNDYMALK
                     LEGGGHYLCEFKSTYKAKKPVRMPGRHEIDRKLDVTSHNRDYTSVEQCEIAIARHSLLG
                     "
     misc_feature    3215..3767
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3359..3376)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3586..3605
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             4020..4489
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4619..4636)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4790..5378)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4870..4889)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5542..6333,6334..6402))
                     /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     6165..6184
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cagctttgct cttaggagtt tcctaataca tcccaaactc aaatatataa agcatttgac
       61 ttgttctatg ccctaggggg cggggggaag ctaagccagc tttttttaac atttaaaatg
      121 ttaattccat tttaaatgca cagatgtttt tatttcataa gggtttcaat gtgcatgaat
      181 gctgcaatat tcctgttacc aaagctagta taaataaaaa tagataaacg tggaaattac
      241 ttagagtttc tgtcattaac gtttccttcc tcagttgaca acataaatgc gctgctgaga
      301 agccagtttg catctgtcag gatcaatttc ccattatgcc agtcatatta attactagtc
      361 aattagttga tttttatttt tgacatatac atgtgaaaga ccccacctgt aggtttggca
      421 agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa
      481 agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg
      541 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc
      601 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc
      661 cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc
      721 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct
      781 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc
      841 gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg
      901 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac
      961 tacccgtcag cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca
     1021 gggaccaccg acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga
     1081 ttgtctagtg tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct
     1141 ctgtatctgg cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg
     1201 gagacgtccc agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg
     1261 atcgttttgg actctttggt gcacccccct tagaggaggg atatgtggtt ctggtaggag
     1321 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa
     1381 gccgcgccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt
     1441 ttctgtattt gtctgaaaat atgggcccgg gccagactgt taccactccc ttaagtttga
     1501 ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga
     1561 agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc ggatggccgc
     1621 gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc ttttcacctg
     1681 gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc ttggcttttg
     1741 acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct cttcctccat
     1801 ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc tccctttatc
     1861 cagccctcac tccttctcta ggcgccccca tatggccata tgagatctta tatggggcac
     1921 ccccgcccct tgtaaacttc cctgaccctg acatgacaag agttactaac agcccctctc
     1981 tccaagctca cttacaggct ctctacttag tccagcacga agtctggaga cctctggcgg
     2041 cagcctacca agaacaactg gaccgaccgg tggtacctca cccttaccga gtcggcgaca
     2101 cagtgtgggt ccgccgacac cagactaaga acctagaacc tcgctggaaa ggaccttaca
     2161 cagtcctgct gaccaccccc accgccctca aagtagacgg catcgcagct tggatacacg
     2221 ccgcccacgt gaaggctgcc gaccccgggg gtggaccatc ctctagactg ccggatccac
     2281 gcgtatgcat ctcgaggcca ccatgtccgt cctgacgccg ctgctgctgc ggggcttgac
     2341 aggctcggcc cggcggctcc cagtgccgcg cgccaagatc cattcgttgc cgccggaggg
     2401 gaagcttggg atgtccgtcc tgacgccgct gctgctgcgg ggcttgacag gctcggcccg
     2461 gcggctccca gtgccgcgcg ccaagatcca ttcgttgccg ccggagggga agctcggggg
     2521 atccgccatg gtgagcgtga tcgccaagca gatgacctac aaggtgtaca tgagcggcac
     2581 cgtgaacggc cactacttcg aggtggaggg cgacggcaag ggcaagccct acgagggcga
     2641 gcagaccgtg aagctgaccg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     2701 gagcccccag ctccagtacg gcagcatccc cttcaccaag taccccgagg acatccccga
     2761 ctacttcaag cagagcttcc ccgagggcta cacctgggag cgcagcatga acttcgagga
     2821 cggcgccgtg tgcaccgtga gcaacgacag cagcatccag ggcaactgct tcatctacaa
     2881 cgtgaagatc agcggcgaga acttcccccc caacggcccc gtgatgcaga agaagaccca
     2941 gggctgggag cccagcaccg agcgcctgtt cgcccgcgac ggaatgctga tcggcaacga
     3001 ctacatggcc ctgaagctgg agggcggcgg ccactacctg tgcgagttca agagcaccta
     3061 caaggccaag aagcccgtga ggatgcccgg ccgccacgag atcgaccgca agctggacgt
     3121 gaccagccac aaccgcgact acaccagcgt ggagcagtgc gagatcgcca tcgcccgcca
     3181 ctccctgctg ggctaagtcg accccccccc cctaacgtta ctggccgaag ccgcttggaa
     3241 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     3301 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     3361 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     3421 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     3481 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     3541 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     3601 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     3661 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     3721 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat
     3781 ggggcgcgcc gtatactgat caatcgattc aattgcaccg gtgcctgcag gccatcccct
     3841 tggcgtctcc tcgacgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     3901 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     3961 gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc
     4021 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     4081 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     4141 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
     4201 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     4261 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     4321 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt
     4381 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     4441 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac atgcagcatg
     4501 tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt
     4561 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4621 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4681 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4741 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4801 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4861 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4921 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4981 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5041 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5101 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5161 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5221 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5281 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5341 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5401 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5461 caaaaaggat cttcacctag atccttttgc ggccggccgc aaatcaatct aaagtatata
     5521 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5581 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5641 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5701 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5761 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5821 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5881 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5941 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6001 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6061 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6121 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6181 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6241 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6301 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6361 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6421 ttattgaagc atttatcagg gttattgtct catga
//
