LOCUS       Exported                6455 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  retrovirus construct for stable expressing mt-mKeima.
ACCESSION   .
VERSION     .
KEYWORDS    pCHAC-mt-mKeima
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
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"
FEATURES             Location/Qualifiers
     source          1..6455
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             2528..3193
                     /codon_start=1
                     /product="human codon-optimized monomeric red fluorescent 
                     protein with a large Stokes shift (Guan et al. 2015)"
                     /label=hmKeima8.5
                     /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(5542..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"