LOCUS       sequence-431711-        7402 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Probe to evaluate lysophagic activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7402)
  AUTHORS   Shima T, Ogura M, Matsuda R, Nakamura S, Jin N, Yoshimori T, Kuma A
  TITLE     The TMEM192-mKeima probe specifically assays lysophagy and reveals 
            its initial steps.
  JOURNAL   J Cell Biol. 2023 Dec 4;222(12):e202204048. doi: 
            10.1083/jcb.202204048. Epub 2023 Oct 6.
  PUBMED    37801070
REFERENCE   2  (bases 1 to 7402)
  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: "J Cell 
            Biol."; date: "2023-12-4"; volume: "222(12):e202204048. doi"; pages:
            " 10.1083/jcb.202204048. Epub 2023 Oct 6"
COMMENT     Sequence Label: pMRX-IB-TMEM192-mKeima
FEATURES             Location/Qualifiers
     source          1..7402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    52..601
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(196..213)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     423..442
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             1205..1674
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(1804..1821)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1975..2563)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2055..2074)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2727..3518,3519..3587))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3350..3369
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3588..3692)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3760..3778
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        3880..4259
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    4618..4975
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5040..5456
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5386..5408
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5424..5440
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    5466..5840
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     5778..5794
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      6728..6737
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6734..6736,6737..6739,6740..7402)
                     /codon_start=1
                     /product="monomeric Keima-Red fluorescent protein, also 
                     known as mKeima"
                     /label=mKeima-Red
                     /translation="MVSVIAKQMTYKVYMSGTVNGHYFEVEGDGKGKPYEGEQTVKLTV
                     TKGGPLPFAWDILSPQLQYGSIPFTKYPEDIPDYFKQSFPEGYTWERSMNFEDGAVCTV
                     SNDSSIQGNCFIYNVKISGENFPPNGPVMQKKTQGWEPSTERLFARDGMLIGNDYMALK
                     LEGGGHYLCEFKSTYKAKKPVRMPGRHEIDRKLDVTSHNRDYTSVEQCEIAIARHSLLG
                     "
ORIGIN
        1 ggccgcaacg gtttccctct agcgggatca attccgcccc ccccccccct aacgttactg
       61 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat
      121 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc
      181 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag
      241 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc
      301 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac
      361 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca
      421 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt
      481 gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa
      541 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgataat
      601 accatggtca tgaaaacatt taacatttct caacaagatc tagaattagt agaagtagcg
      661 acagagaaga ttacaatgct ttatgaggat aataaacatc atgtgggagc ggcaattcgt
      721 acgaaaacag gagaaatcat ttcggcagta catattgaag cgtatatagg acgagtaact
      781 gtttgtgcag aagccattgc gattggtagt gcagtttcga atggacaaaa ggattttgac
      841 acgattgtag ctgttagaca cccttattct gacgaagtag atagaagtat tcgagtggta
      901 agtccttgtg gtatgtgtag ggagttgatt tcagactatg caccagattg ttttgtgtta
      961 atagaaatga atggcaagtt agtcaaaact acgattgaag aactcattcc actcaaatat
     1021 acccgaaatt aaaagtcgac gataaaataa aagattttat ttagtctcca gaaaaagggg
     1081 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     1141 gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg aacagatgga
     1201 acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     1261 ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc
     1321 tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc tcagcagttt
     1381 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     1441 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     1501 caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgattga ctgagtcgcc
     1561 cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt
     1621 tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcacatgca
     1681 gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat ggccatagtt
     1741 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     1801 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     1861 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     1921 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1981 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2041 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2101 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2161 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2221 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2281 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2341 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2401 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2461 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2521 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2581 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2641 attatcaaaa aggatcttca cctagatcct tttgcggccg gccgcaaatc aatctaaagt
     2701 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2761 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     2821 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     2881 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     2941 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3001 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3061 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3121 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3181 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3241 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3301 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg
     3361 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3421 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3481 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3541 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3601 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3661 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     3721 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     3781 tttcgtcttc actcgaggag cttggcccat tgcatacgtt gtatccatat cataatatgt
     3841 acatttatat tggctcatgt ccaacattac cgccatgttg acattgatta ttgactagtt
     3901 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     3961 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     4021 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     4081 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     4141 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     4201 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc
     4261 tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat
     4321 ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc
     4381 aataaaagag cccacaaccc ctcactcggc gcgccagtcc tccgattgac tgagtcgccc
     4441 gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt
     4501 ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt tcatttgggg
     4561 gctcgtccgg gatcgggaga cccctgccca gggaccaccg acccaccacc gggaggtaag
     4621 ctggccagca acttatctgt gtctgtccga ttgtctagtg tctatgactg attttatgcg
     4681 cctgcgtcgg tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga
     4741 cgagttcgga acacccggcc gcaaccctgg gagacgtccc agggacttcg ggggccgttt
     4801 ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg actctttggt gcacccccct
     4861 tagaggaggg atatgtggtt ctggtaggag acgagaacct aaaacagttc ccgcctccgt
     4921 ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc gcgtcttgtc tgctgcagca
     4981 tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt gtctgaaaat atgggcccgg
     5041 gccagactgt taccactccc ttaagtttga ccttaggtca ctggaaagat gtcgagcgga
     5101 tcgctcacaa ccagtcggta gatgtcaaga agagacgttg ggttaccttc tgctctgcag
     5161 aatggccaac ctttaacgtc ggatggccgc gagacggcac ctttaaccga gacctcatca
     5221 cccaggttaa gatcaaggtc ttttcacctg gcccgcatgg acacccagac caggtcccct
     5281 acatcgtgac ctgggaagcc ttggcttttg acccccctcc ctgggtcaag ccctttgtac
     5341 accctaagcc tccgcctcct cttcctccat ccgccccgtc tctccccctt gaacctcctc
     5401 gttcgacccc gcctcgatcc tccctttatc cagccctcac tccttctcta ggcgccccca
     5461 tatggccata tgagatctta tatggggcac ccccgcccct tgtaaacttc cctgaccctg
     5521 acatgacaag agttactaac agcccctctc tccaagctca cttacaggct ctctacttag
     5581 tccagcacga agtctggaga cctctggcgg cagcctacca agaacaactg gaccgaccgg
     5641 tggtacctca cccttaccga gtcggcgaca cagtgtgggt ccgccgacac cagactaaga
     5701 acctagaacc tcgctggaaa ggaccttaca cagtcctgct gaccaccccc accgccctca
     5761 aagtagacgg catcgcagct tggatacacg ccgcccacgt gaaggctgcc gaccccgggg
     5821 gtggaccatc ctctagactg ccggatccca gtgtggtggt acgggaatta tggcggcggg
     5881 gggcaggatg gaggacggtt ccttggatat cacccagagt attgaagacg acccacttct
     5941 ggatgcccag cttctcccac accactcatt acaagctcac tttagacccc gattccatcc
     6001 tcttcctaca gtcatcatag tgaatcttct gtggtttatt catctcgtgt ttgttgtttt
     6061 agcattttta acaggtgtgc tttgttctta tcctaatcca aatgaggaca agtgcccagg
     6121 aaattacaca aacccattga aagttcagac ggttataatc cttgggaaag ttattttgtg
     6181 gattctccat ttactccttg aatgctacat ccagtatcac cacagcaaaa tcagaaaccg
     6241 aggctataac ttgatctacc gatcaacaag gcatctcaag agacttgcgt tgatgataca
     6301 gtcctctggc aacacagtgc ttctcctcat actgtgcatg cagcactcct tcccagagcc
     6361 tggcagattg tatcttgacc tcattctggc catcttggca ctggaactca tctgttccct
     6421 gatatgtctc ctcatttaca cagtgaaaat ccggagattt aataaagcta aaccagagcc
     6481 tgatatactt gaagaagaaa aaatctatgc ttaccccagc aatattacct cggagactgg
     6541 attcagaact atttcaagcc tagaagaaat tgttgaaaag caaggagaca ccattgaata
     6601 cctgaagcga cacaatgcgc tgctgagtaa gcgattgttg gctctcactt cctcagacct
     6661 gggctgtcag ccaagtagaa cgggaattct gcagtcgacg gtaccgcggg cccgggatcc
     6721 accggtcgcc accatggtga gtgtgatcgc taaacaaatg acctacaagg tttatatgtc
     6781 aggcacggtc aatggacact actttgaggt cgaaggcgat ggaaaaggaa agccttacga
     6841 gggagagcag acagtaaagc tcactgtcac caagggtgga cctctgccat ttgcttggga
     6901 tattttatca ccacagcttc agtacggaag cataccattc accaagtacc ctgaagacat
     6961 ccctgattat ttcaagcagt cattccctga gggatataca tgggagagga gcatgaactt
     7021 tgaagatggt gcagtgtgta ctgtcagcaa tgattccagc atccaaggca actgtttcat
     7081 ctacaatgtc aaaatctctg gtgagaactt tcctcccaat ggacctgtta tgcagaagaa
     7141 gacacagggc tgggaaccca gcactgagcg tctctttgca cgagatggaa tgctgatagg
     7201 aaacgattat atggctctga agttggaagg aggtggtcac tatttgtgtg aatttaaatc
     7261 tacttacaag gcaaagaagc ctgtgaggat gccagggcgc cacgagattg accgcaaact
     7321 ggatgtaacc agtcacaaca gggattacac atctgttgag cagtgtgaaa tagccattgc
     7381 acgccactct ttgctcggtt aa
//
