LOCUS       sequence-364671-        5917 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Lamp1 fused to YFP. Used for localization to
            the lysosome..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5917)
  AUTHORS   Sherer NM, Lehmann MJ, Jimenez-Soto LF, Ingmundson A, Horner SM, 
            Cicchetti G, Allen PG, Pypaert M, Cunningham JM, Mothes W
  TITLE     Visualization of retroviral replication in living cells reveals 
            budding into multivesicular bodies.
  JOURNAL   Traffic 2003 Nov;4(11):785-801.
  PUBMED    14617360
REFERENCE   2  (bases 1 to 5917)
  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: "Traffic"; 
            date: "2003-11"; volume: "4(11)"; pages: "785-801"
COMMENT     Sequence Label: Lamp1-YFP
FEATURES             Location/Qualifiers
     source          1..5917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        38..341
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        342..545
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     496..516
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      614..623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      618..627
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(620..682,683..1840)
                     /codon_start=1
                     /product="rat lysosome-associated membrane glycoprotein 1"
                     /label=LAMP1
                     /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF
                     LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT
                     RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI
                     VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT
                     CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG
                     MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL
                     ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR
                     SHAGYQTI"
     regulatory      1856..1865
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1862..1864,1865..1867,1868..2581)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1907..1928)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2168..2187)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2515..2536
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2704..2825
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2741..2760)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2795..2814
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2832..3287)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2969..2990)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3181..3200
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3314..3418
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3420..3611
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3443..3463)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3608..3741
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3790..4584
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3790..4584
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3844..3863)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4454..4473
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4772..4791)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4816..4863
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5192..5780
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5681..5700
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
       61 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      121 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      181 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      241 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      301 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      361 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      421 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      481 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      541 gagctggttt agtgaaccgt cagatccgct agcgctaccg gactcagatc tcgagctcaa
      601 gcttcgaatt ctcgccacca tggcggcccc gggcgcccgg cggccgctgc tcctgttgct
      661 gctggcaggc cttgcacaca gcgccccagc actgttcgag gtgaaagaca acaacggcac
      721 agcgtgtata atggccagct tctctgcctc ctttctgacc acctatgagg ctggacatgt
      781 ttctaaggtc tcgaatatga ccctgccagc ctctgcagaa gtcctgaaga atagcagctc
      841 ttgtggtgaa aagaatgctt ctgagcccac cctcgcaatc acctttggag aaggatattt
      901 actgaaactc accttcacaa aaaacacaac acgttacagt gtccagcaca tgtatttcac
      961 atataacctg tcagacacac aattctttcc caatgccagc tccaaagggc ccgacactgt
     1021 ggattccaca actgacatca aggcagacat caacaaaaca taccgatgtg tcagcgacat
     1081 cagggtctac atgaagaatg tgaccattgt gctctgggac gctactatcc aggcctacct
     1141 gccgagtagc aacttcagca aggaagagac acgctgccca caggatcaac cttccccaac
     1201 tactgggcca cccagcccct caccaccact cgtgcccaca aaccccagtg tgtccaagta
     1261 caatgtgact ggtgacaatg gaacctgcct gctggcctct atggcactgc aactcaacat
     1321 cacctacatg aagaaggaca acacgactgt gaccagagca ttcaacatca acccaagtga
     1381 caaatatagt gggacttgcg gtgcccagtt ggtgaccctg aaggtgggga acaagagcag
     1441 agtcctggag ctgcagtttg ggatgaatgc cacttctagc ctgtttttcc tgcaaggagt
     1501 tcagttgaac atgactcttc ctgatgccat agagcccacg ttcagcacct ccaactattc
     1561 cctgaaagct cttcaggcca gtgtcggcaa ctcatacaag tgcaacagtg aggagcacat
     1621 ctttgtcagc aaggcgctcg ccctcaatgt cttcagcgtg caagtccagg ctttcagggt
     1681 agaaagtgac aggtttgggt ctgtggaaga gtgtgtacag gacggtaaca acatgctgat
     1741 ccccattgct gtgggcgggg ccctggcagg gctggtcctc atcgtcctca tcgcctacct
     1801 catcggcagg aagaggagtc acgcgggcta tcagaccatc cgggatccac cggtcgccac
     1861 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1921 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1981 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2041 cctcgtgacc accttcggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa
     2101 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2161 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2221 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2281 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     2341 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     2401 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2461 ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     2521 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     2581 aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
     2641 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     2701 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2761 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2821 tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2881 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2941 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     3001 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     3061 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     3121 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3181 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     3241 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     3301 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3361 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     3421 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3481 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3541 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3601 accatagtcc cgcccctaac tccgcccatc ccgcccattc tccgccccat ggctgactaa
     3661 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3721 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     3781 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3841 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3901 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3961 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4021 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4081 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4141 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4201 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4261 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     4321 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4381 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4441 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4501 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4561 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4621 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4681 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4741 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     4801 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     4861 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     4921 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     4981 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     5041 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5101 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5161 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5221 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5281 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5341 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5401 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5461 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5521 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5581 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5641 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5701 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5761 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5821 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5881 ttaccgccat gcattagtta ttaatagtaa tcaatta
//
