LOCUS       sequence-443442-        7880 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of LAMP1-mRuby3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7880)
  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 7880)
  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: pMRX-IZU-LAMP1-mRuby3
FEATURES             Location/Qualifiers
     source          1..7880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    739..1096
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1161..1577
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1507..1529
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1545..1561
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1587..1961
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1899..1915
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1988..1997
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1992..2001
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1994..2077,2078..3244)
                     /codon_start=1
                     /product="human lysosome-associated membrane glycoprotein 
                     1"
                     /label=LAMP1
                     /translation="MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMAN
                     FSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNF
                     TRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHM
                     NNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKY
                     NVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSE
                     GTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNA
                     EEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLIV
                     LIAYLVGRKRSHAGYQTI"
     misc_feature    4016..4582
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4032..4584
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4176..4193)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4403..4422
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4602..4976
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     LTR             5206..5675
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5805..5822)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5976..6564)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6056..6075)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6728..7519,7520..7588))
                     /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     7351..7370
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7589..7693)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7761..7779
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg ctagagaacc atcagatgtt tccagggtgc cccaaggacc
      421 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
      481 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
      541 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
      601 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
      661 gcgggggtct ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc
      721 gacccaccac cgggaggtaa gctggccagc aacttatctg tgtctgtccg attgtctagt
      781 gtctatgact gattttatgc gcctgcgtcg gtactagtta gctaactagc tctgtatctg
      841 gcggacccgt ggtggaactg acgagttcgg aacacccggc cgcaaccctg ggagacgtcc
      901 cagggacttc gggggccgtt tttgtggccc gacctgagtc caaaaatccc gatcgttttg
      961 gactctttgg tgcacccccc ttagaggagg gatatgtggt tctggtagga gacgagaacc
     1021 taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttgggaccga agccgcgccg
     1081 cgcgtcttgt ctgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt
     1141 tgtctgaaaa tatgggcccg ggccagactg ttaccactcc cttaagtttg accttaggtc
     1201 actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt
     1261 gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca
     1321 cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg
     1381 gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc
     1441 cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt
     1501 ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca
     1561 ctccttctct aggcgccccc atatggccat atgagatctt atatggggca cccccgcccc
     1621 ttgtaaactt ccctgaccct gacatgacaa gagttactaa cagcccctct ctccaagctc
     1681 acttacaggc tctctactta gtccagcacg aagtctggag acctctggcg gcagcctacc
     1741 aagaacaact ggaccgaccg gtggtacctc acccttaccg agtcggcgac acagtgtggg
     1801 tccgccgaca ccagactaag aacctagaac ctcgctggaa aggaccttac acagtcctgc
     1861 tgaccacccc caccgccctc aaagtagacg gcatcgcagc ttggatacac gccgcccacg
     1921 tgaaggctgc cgaccccggg ggtggaccat cctctagact gccggatctg ccatcgatat
     1981 aaagcttgcc accatggcgg cccccggcag cgcccggcga cccctgctgc tgctactgct
     2041 gttgctgctg ctcggcctca tgcattgtgc gtcagcagca atgtttatgg tgaaaaatgg
     2101 caacgggacc gcgtgcataa tggccaactt ctctgctgcc ttctcagtga actacgacac
     2161 caagagtggc cctaagaaca tgacctttga cctgccatca gatgccacag tggtgctcaa
     2221 ccgcagctcc tgtggaaaag agaacacttc tgaccccagt ctcgtgattg cttttggaag
     2281 aggacataca ctcactctca atttcacgag aaatgcaaca cgttacagcg tccagctcat
     2341 gagttttgtt tataacttgt cagacacaca ccttttcccc aatgcgagct ccaaagaaat
     2401 caagactgtg gaatctataa ctgacatcag ggcagatata gataaaaaat acagatgtgt
     2461 tagtggcacc caggtccaca tgaacaacgt gaccgtaacg ctccatgatg ccaccatcca
     2521 ggcgtacctt tccaacagca gcttcagcag gggagagaca cgctgtgaac aagacaggcc
     2581 ttccccaacc acagcgcccc ctgcgccacc cagcccctcg ccctcacccg tgcccaagag
     2641 cccctctgtg gacaagtaca acgtgagcgg caccaacggg acctgcctgc tggccagcat
     2701 ggggctgcag ctgaacctca cctatgagag gaaggacaac acgacggtga caaggcttct
     2761 caacatcaac cccaacaaga cctcggccag cgggagctgc ggcgcccacc tggtgactct
     2821 ggagctgcac agcgagggca ccaccgtcct gctcttccag ttcgggatga atgcaagttc
     2881 tagccggttt ttcctacaag gaatccagtt gaatacaatt cttcctgacg ccagagaccc
     2941 tgcctttaaa gctgccaacg gctccctgcg agcgctgcag gccacagtcg gcaattccta
     3001 caagtgcaac gcggaggagc acgtccgtgt cacgaaggcg ttttcagtca atatattcaa
     3061 agtgtgggtc caggctttca aggtggaagg tggccagttt ggctctgtgg aggagtgtct
     3121 gctggacgag aacagcatgc tgatccccat cgctgtgggt ggtgccctgg cggggctggt
     3181 cctcatcgtc ctcatcgcct acctcgtcgg caggaagagg agtcacgcag gctaccagac
     3241 tatcggatcc gggaattcgg gctctggttc aggcgtgagc aagggcgagg agctcatcaa
     3301 ggagaacatg cggatgaaag tcgtcatgga aggctctgtc aatgggcacc agttcaagtg
     3361 taccggagaa ggggaagggc gtccctatga aggcgtacaa accatgcgca tcaaagtgat
     3421 agagggcggg ccactccctt ttgccttcga cattctggct acctccttca tgtatggcag
     3481 tcggacattc atcaagtatc ccgcagacat tccggacttc ttcaaacagt cctttccaga
     3541 aggctttaca tgggagaggg tgactaggta cgaagatggc ggagtggtta ctgtgactca
     3601 ggacacgtca ttggaggacg gagagctggt gtacaatgtg aaagtacgcg gagtgaactt
     3661 tcccagcaat ggtcctgtta tgcagaagaa aaccaagggt tgggaaccca atacggagat
     3721 gatgtatcca gctgatggtg gcctgagggg atacacagac atagccctga aagtcgatgg
     3781 aggtggccat ctgcactgca actttgtgac cacctacaga agcaagaaaa ctgtggggaa
     3841 cattaagatg cctggagtgc atgcagtcga tcacagactt gagagaatcg aggagtctga
     3901 taacgagaca tacgtggtac aacgagaagt tgccgttgcg aagtacagca atctgggcgg
     3961 gggcatggac gagctgtaca agtaatgatt tctcgagagc ggccgctacg taaattccgc
     4021 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     4081 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     4141 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     4201 gttgaatgtc gtgaaggaag cagttcctct ggaagtttct tgaagacaaa caacgtctgt
     4261 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     4321 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     4381 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     4441 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
     4501 atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     4561 cctttgaaaa acacgatgat aatatggcca caaccgtcga catggccaag ttgaccagtg
     4621 ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc
     4681 tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga
     4741 ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt
     4801 gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc
     4861 gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg
     4921 ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgagtcg
     4981 accgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catggctcga
     5041 cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     5101 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac
     5161 tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa
     5221 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag
     5281 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     5341 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt
     5401 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     5461 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     5521 ccctcactcg gcgcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
     5581 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     5641 tgagtgattg actacccgtc agcgggggtc tttcacatgc agcatgtatc aaaattaatt
     5701 tggttttttt tcttaagtat ttacattaaa tggccatagt tgcattaatg aatcggccaa
     5761 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5821 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5881 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5941 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6001 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6061 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6121 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6181 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6241 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6301 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6361 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     6421 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6481 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6541 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6601 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6661 acctagatcc ttttgcggcc ggccgcaaat caatctaaag tatatatgag taaacttggt
     6721 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6781 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6841 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6901 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6961 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7021 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7081 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7141 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7201 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7261 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7321 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
     7381 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7441 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7501 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7561 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7621 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7681 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     7741 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt cactatagga
     7801 gcttggccca ttgcatacgt tgtatccata tcataatatg tacatttata ttggctcatg
     7861 tccaacatta ccgccatgtt
//
