LOCUS       sequence-443149-        6991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of mRuby3-LC3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6991)
  AUTHORS   Shinoda S, Sakai Y, Matsui T, Uematsu M, Koyama-Honda I, Sakamaki 
            JI, Yamamoto H, Mizushima N
  TITLE     Syntaxin 17 recruitment to mature autophagosomes is temporally 
            regulated by PI4P accumulation.
  JOURNAL   Elife. 2024 Jun 4;12:RP92189. doi: 10.7554/eLife.92189.
  PUBMED    38831696
REFERENCE   2  (bases 1 to 6991)
  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: "Elife."; 
            date: "2024-06-4"; volume: "12:RP92189. doi"; pages: " 
            10.7554/eLife.92189"
COMMENT     Sequence Label: pMRXIB-mRuby3-LC3
FEATURES             Location/Qualifiers
     source          1..6991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    667..1024
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1089..1505
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1435..1457
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1473..1489
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1515..1889
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1827..1843
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1916..1925
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3055..3621
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3071..3623
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3215..3232)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3442..3461
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             4245..4714
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4844..4861)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5015..5603)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5095..5114)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5767..6558,6559..6627))
                     /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     6390..6409
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6628..6732)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6800..6818
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(6920..6991,1..308)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
       61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      301 ttaccatgct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
      361 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
      421 cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgattgact
      481 gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt
      541 ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt
      601 catttggggg ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg
      661 ggaggtaagc tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgactga
      721 ttttatgcgc ctgcgtcggt actagttagc taactagctc tgtatctggc ggacccgtgg
      781 tggaactgac gagttcggaa cacccggccg caaccctggg agacgtccca gggacttcgg
      841 gggccgtttt tgtggcccga cctgagtcca aaaatcccga tcgttttgga ctctttggtg
      901 cacccccctt agaggaggga tatgtggttc tggtaggaga cgagaaccta aaacagttcc
      961 cgcctccgtc tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct
     1021 gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaata
     1081 tgggcccggg ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg
     1141 tcgagcggat cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct
     1201 gctctgcaga atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag
     1261 acctcatcac ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc
     1321 aggtccccta catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc
     1381 cctttgtaca ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg
     1441 aacctcctcg ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag
     1501 gcgcccccat atggccatat gagatcttat atggggcacc cccgcccctt gtaaacttcc
     1561 ctgaccctga catgacaaga gttactaaca gcccctctct ccaagctcac ttacaggctc
     1621 tctacttagt ccagcacgaa gtctggagac ctctggcggc agcctaccaa gaacaactgg
     1681 accgaccggt ggtacctcac ccttaccgag tcggcgacac agtgtgggtc cgccgacacc
     1741 agactaagaa cctagaacct cgctggaaag gaccttacac agtcctgctg accaccccca
     1801 ccgccctcaa agtagacggc atcgcagctt ggatacacgc cgcccacgtg aaggctgccg
     1861 accccggggg tggaccatcc tctagactgc cggatctgcc atcgatataa agcttgccac
     1921 catggtgagc aagggcgagg agctcatcaa ggagaacatg cggatgaaag tcgtcatgga
     1981 aggctctgtc aatgggcacc agttcaagtg taccggagaa ggggaagggc gtccctatga
     2041 aggcgtacaa accatgcgca tcaaagtgat agagggcggg ccactccctt ttgccttcga
     2101 cattctggct acctccttca tgtatggcag tcggacattc atcaagtatc ccgcagacat
     2161 tccggacttc ttcaaacagt cctttccaga aggctttaca tgggagaggg tgactaggta
     2221 cgaagatggc ggagtggtta ctgtgactca ggacacgtca ttggaggacg gagagctggt
     2281 gtacaatgtg aaagtacgcg gagtgaactt tcccagcaat ggtcctgtta tgcagaagaa
     2341 aaccaagggt tgggaaccca atacggagat gatgtatcca gctgatggtg gcctgagggg
     2401 atacacagac atagccctga aagtcgatgg aggtggccat ctgcactgca actttgtgac
     2461 cacctacaga agcaagaaaa ctgtggggaa cattaagatg cctggagtgc atgcagtcga
     2521 tcacagactt gagagaatcg aggagtctga taacgagaca tacgtggtac aacgagaagt
     2581 tgccgttgcg aagtacagca atctgggcgg gggcatggac gagctgtaca agggttcagg
     2641 ctcgggatcc gggaattcga tgccgtccga gaagaccttc aaacagcgcc ggagcttcga
     2701 acaaagagtg gaagatgtcc ggctcatccg ggagcagcac cccaccaaga tcccagtgat
     2761 tatagagcga tacaagggtg agaagcagct gcccgtcctg gacaagacca agttccttgt
     2821 acctgatcac gtgaatatga gcgaactcat caagataatt agaaggcgcc tgcagctcaa
     2881 tgctaaccaa gccttcttcc tcctggtgaa tgggcacagc atggtgagtg tgtccacacc
     2941 catctctgaa gtgtacgaga gcgagagaga tgaagacggc ttcctgtaca tggtctatgc
     3001 ctcccaggag acgttcggga cagcactggc tgtttgagcg gccgctacgt aaattccgcc
     3061 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     3121 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     3181 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     3241 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     3301 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     3361 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     3421 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     3481 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     3541 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     3601 ctttgaaaaa cacgatgata atatggccac aaccatggtc atgaaaacat ttaacatttc
     3661 tcaacaagat ctagaattag tagaagtagc gacagagaag attacaatgc tttatgagga
     3721 taataaacat catgtgggag cggcaattcg tacgaaaaca ggagaaatca tttcggcagt
     3781 acatattgaa gcgtatatag gacgagtaac tgtttgtgca gaagccattg cgattggtag
     3841 tgcagtttcg aatggacaaa aggattttga cacgattgta gctgttagac acccttattc
     3901 tgacgaagta gatagaagta ttcgagtggt aagtccttgt ggtatgtgta gggagttgat
     3961 ttcagactat gcaccagatt gttttgtgtt aatagaaatg aatggcaagt tagtcaaaac
     4021 tacgattgaa gaactcattc cactcaaata tacccgaaat taacgacccc atggctcgac
     4081 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg
     4141 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact
     4201 gagaatagag aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa
     4261 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc
     4321 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     4381 aacagatggt ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt
     4441 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
     4501 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
     4561 cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat
     4621 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct
     4681 gagtgattga ctacccgtca gcgggggtct ttcacatgca gcatgtatca aaattaattt
     4741 ggtttttttt cttaagtatt tacattaaat ggccatagtt gcattaatga atcggccaac
     4801 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4861 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4921 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4981 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5041 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5101 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5161 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5221 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5281 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5341 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5401 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5461 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5521 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5581 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5641 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5701 cctagatcct tttgcggccg gccgcaaatc aatctaaagt atatatgagt aaacttggtc
     5761 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5821 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5881 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5941 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6001 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6061 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6121 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6181 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6241 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6301 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6361 gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa
     6421 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6481 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6541 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6601 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6661 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6721 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     6781 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc actcgaggag
     6841 cttggcccat tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt
     6901 ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg
     6961 gggtcattag ttcatagccc atatatggag t
//
