LOCUS       sequence-209422-        8078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Lamp1 tagged with Venus in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8078)
  AUTHORS   Tsuboyama K, Koyama-Honda I, Sakamaki Y, Koike M, Morishita H, 
            Mizushima N
  TITLE     The ATG conjugation systems are important for degradation of the 
            inner autophagosomal membrane.
  JOURNAL   Science. 2016 Nov 25;354(6315):1036-1041. Epub 2016 Oct 20.
  PUBMED    27885029
REFERENCE   2  (bases 1 to 8078)
  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: "Science."; 
            date: "2016-11-25"; volume: "354(6315)"; pages: "1036-1041. Epub 
            2016 Oct 20"
COMMENT     Sequence Label: pMRXIP Lamp1-Venus
FEATURES             Location/Qualifiers
     source          1..8078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..358
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             423..839
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     769..791
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     807..823
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    849..1223
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1161..1177
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1244..1253
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1246..1308,1309..2466)
                     /codon_start=1
                     /product="rat lysosome-associated membrane glycoprotein 1"
                     /label=LAMP1
                     /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF
                     LTTYDAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT
                     RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI
                     VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT
                     CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG
                     MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL
                     ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR
                     SHAGYQTI"
     CDS             join(2506..2508,2509..2511,2512..3225)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2551..2572)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2812..2831)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3159..3180
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(3256..3830,3831..3833,3834..3842)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3423..3440)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3650..3669
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3843..4442
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3843..3862)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4339..4359
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4666..5135
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5265..5282)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5436..6024)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5516..5535)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6188..6979,6980..7048))
                     /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     6811..6830
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7049..7153)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7221..7239
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        7341..7720
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 aagctggcca gcaacttatc tgtgtctgtc cgattgtcta gtgtctatga ctgattttat
       61 gcgcctgcgt cggtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac
      121 tgacgagttc ggaacacccg gccgcaaccc tgggagacgt cccagggact tcgggggccg
      181 tttttgtggc ccgacctgag tccaaaaatc ccgatcgttt tggactcttt ggtgcacccc
      241 ccttagagga gggatatgtg gttctggtag gagacgagaa cctaaaacag ttcccgcctc
      301 cgtctgaatt tttgctttcg gtttgggacc gaagccgcgc cgcgcgtctt gtctgctgca
      361 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aatatgggcc
      421 cgggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc
      481 ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg
      541 cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca
      601 tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc
      661 cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg
      721 tacaccctaa gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc
      781 ctcgttcgac cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccc
      841 ccatatggcc atatgagatc ttatatgggg cacccccgcc ccttgtaaac ttccctgacc
      901 ctgacatgac aagagttact aacagcccct ctctccaagc tcacttacag gctctctact
      961 tagtccagca cgaagtctgg agacctctgg cggcagccta ccaagaacaa ctggaccgac
     1021 cggtggtacc tcacccttac cgagtcggcg acacagtgtg ggtccgccga caccagacta
     1081 agaacctaga acctcgctgg aaaggacctt acacagtcct gctgaccacc cccaccgccc
     1141 tcaaagtaga cggcatcgca gcttggatac acgccgccca cgtgaaggct gccgaccccg
     1201 ggggtggacc atcctctaga ctgccggatc atcgatgaat tcgccatggc ggccccgggc
     1261 gcccggcggc cgctgctcct gttgctgctg gcaggccttg cacacagcgc cccagcactg
     1321 ttcgaggtga aagacaacaa cggcacagcg tgtataatgg ccagcttctc tgcctccttt
     1381 ctgaccacct atgatgctgg acatgtttct aaggtctcga atatgaccct gccagcctct
     1441 gcagaagtcc tgaagaatag cagctcttgt ggtgaaaaga atgcttctga gcccaccctc
     1501 gcaatcacct ttggagaagg atatttactg aaactcacct tcacaaaaaa cacaacacgt
     1561 tacagtgtcc agcacatgta tttcacatat aacctgtcag acacacaatt ctttcccaat
     1621 gccagctcca aagggcccga cactgtggat tccacaactg acatcaaggc agacatcaac
     1681 aaaacatacc gatgtgtcag cgacatcagg gtctacatga agaatgtgac cattgtgctc
     1741 tgggacgcta ctatccaggc ctacctgccg agtagcaact tcagcaagga agagacacgc
     1801 tgcccacagg atcaaccttc cccaactact gggccaccca gcccctcacc accacttgtg
     1861 cccacaaacc ccagtgtgtc caagtacaat gtgactggtg acaatggaac ctgcctgctg
     1921 gcctctatgg cactgcaact caacatcacc tacatgaaga aggacaacac gactgtgacc
     1981 agagcattca acatcaaccc aagtgacaaa tatagtggga cttgcggtgc ccagttggtg
     2041 accctgaagg tggggaacaa gagcagagtc ctggagctgc agtttgggat gaatgccact
     2101 tctagcctgt ttttcctgca aggagttcag ttgaacatga ctcttcctga tgccatagag
     2161 cccacgttca gcacctccaa ctattccctg aaagctcttc aggccagtgt cggcaactca
     2221 tacaagtgca acagtgagga gcacatcttt gtcagcaagg cgctcgccct caatgtcttc
     2281 agcgtgcaag tccaggcttt cagggtagaa agtgacaggt ttgggtctgt ggaagagtgt
     2341 gtacaggacg gtaacaacat gctgatcccc attgctgtgg gcggggccct ggcagggctg
     2401 gtcctcatcg tcctcatcgc ctacctcatc ggcaggaaga ggagtcacgc gggctatcag
     2461 accatcgaat tcactagtgt taacggatcc ctcgagcggc cgcccatggt gagcaagggc
     2521 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2581 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     2641 aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2701 ggctacggcc tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     2761 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     2821 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2881 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2941 tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     3001 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     3061 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     3121 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     3181 accgccgccg ggatcactct cggcatggac gagctgtaca agtaagatgg ccgctacgta
     3241 aattccgccc ccccccccct ctccctcccc cccccctaac gttactggcc gaagccgctt
     3301 ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg
     3361 caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc
     3421 ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga
     3481 agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc
     3541 tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc
     3601 acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc
     3661 aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga
     3721 tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg
     3781 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa
     3841 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg
     3901 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg
     3961 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     4021 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     4081 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     4141 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     4201 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     4261 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     4321 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     4381 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     4441 gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catggctcga
     4501 cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     4561 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac
     4621 tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa
     4681 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag
     4741 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     4801 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt
     4861 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     4921 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     4981 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
     5041 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     5101 tgagtgattg actacccgtc agcgggggtc tttcacatgc agcatgtatc aaaattaatt
     5161 tggttttttt tcttaagtat ttacattaaa tggccatagt tgcattaatg aatcggccaa
     5221 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5281 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5341 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5401 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5461 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5521 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5581 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5641 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5701 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5761 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5821 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5881 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5941 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6001 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6061 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6121 acctagatcc ttttgcggcc ggccgcaaat caatctaaag tatatatgag taaacttggt
     6181 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6241 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6301 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6361 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6421 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6481 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6541 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6601 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6661 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6721 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6781 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
     6841 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6901 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6961 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7021 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7081 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7141 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     7201 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt cactcgagga
     7261 gcttggccca ttgcatacgt tgtatccata tcataatatg tacatttata ttggctcatg
     7321 tccaacatta ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac
     7381 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     7441 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     7501 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     7561 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     7621 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     7681 ttggcagtac atctacgtat tagtcatcgc tattaccatg ctagagaacc atcagatgtt
     7741 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
     7801 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
     7861 cctcactcgg cgcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat
     7921 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct
     7981 gagtgattga ctacccgtca gcgggggtct ttcatttggg ggctcgtccg ggatcgggag
     8041 acccctgccc agggaccacc gacccaccac cgggaggt
//
