LOCUS       sequence-439309-        5951 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Lysosome Membrane, Excitation: 549, Emission: 565.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5951)
  AUTHORS   Shaner NC, Lin MZ, McKeown MR, Steinbach PA, Hazelwood KL, Davidson 
            MW, Tsien RY
  TITLE     Improving the photostability of bright monomeric orange and red 
            fluorescent proteins.
  JOURNAL   Nat Methods. 2008 Jun;5(6):545-51. doi: 10.1038/nmeth.1209. Epub 
            2008 May 4.
  PUBMED    18454154
REFERENCE   2  (bases 1 to 5951)
  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"; doi: 
            "10.1038/nmeth.1209"; journalName: "Nat Methods"; date: "2008-06"; 
            volume: "5"; issue: "6"; pages: "545-51"
COMMENT     Sequence Label: mOrange2-Lysosomes-20
FEATURES             Location/Qualifiers
     source          1..5951
                     /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      597..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      601..610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(603..665,666..1823)
                     /codon_start=1
                     /product="rat lysosome-associated membrane glycoprotein 1"
                     /label=LAMP1
                     /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF
                     LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT
                     RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI
                     VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT
                     CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG
                     MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL
                     ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR
                     SHAGYQTI"
     regulatory      1878..1887
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1884..2594
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     polyA_signal    2716..2837
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2753..2772)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2807..2826
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2844..3299)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2981..3002)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3193..3212
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3326..3430
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3432..3789
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3432..3623
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3455..3475)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3640..3775
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3824..4618
                     /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             3824..4618
                     /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(3878..3897)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4488..4507
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4806..4825)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4850..4897
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5226..5814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5715..5734
                     /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 tcgagcgcca
      601 ccatggcggc cccgggcgcc cggcggccgc tgctcctgtt gctgctggca ggccttgcac
      661 acagcgcccc agcactgttc gaggtgaaag acaacaacgg cacagcgtgt ataatggcca
      721 gcttctctgc ctcctttctg accacctatg aggctggaca tgtttctaag gtctcgaata
      781 tgaccctgcc agcctctgca gaagtcctga agaatagcag ctcttgtggt gaaaagaatg
      841 cttctgagcc caccctcgca atcacctttg gagaaggata tttactgaaa ctcaccttca
      901 caaaaaacac aacacgttac agtgtccagc acatgtattt cacatataac ctgtcagaca
      961 cacaattctt tcccaatgcc agctccaaag ggcccgacac tgtggattcc acaactgaca
     1021 tcaaggcaga catcaacaaa acataccgat gtgtcagcga catcagggtc tacatgaaga
     1081 atgtgaccat tgtgctctgg gacgctacta tccaggccta cctgccgagt agcaacttca
     1141 gcaaggaaga gacacgctgc ccacaggatc aaccttcccc aactactggg ccacccagcc
     1201 cctcaccacc acttgtgccc acaaacccca gtgtgtccaa gtacaatgtg actggtgaca
     1261 atggaacctg cctgctggcc tctatggcac tgcaactcaa catcacctac atgaagaagg
     1321 acaacacgac tgtgaccaga gcattcaaca tcaacccaag tgacaaatat agtgggactt
     1381 gcggtgccca gttggtgacc ctgaaggtgg ggaacaagag cagagtcctg gagctgcagt
     1441 ttgggatgaa tgccacttct agcctgtttt tcctgcaagg agttcagttg aacatgactc
     1501 ttcctgatgc catagagccc acgttcagca cctccaacta ttccctgaaa gctcttcagg
     1561 ccagtgtcgg caactcatac aagtgcaaca gtgaggagca catctttgtc agcaaggcgc
     1621 tcgccctcaa tgtcttcagc gtgcaagtcc aggctttcag ggtagaaagt gacaggtttg
     1681 ggtctgtgga agagtgtgta caggacggta acaacatgct gatccccatt gctgtgggcg
     1741 gggccctggc agggctggtc ctcatcgtcc tcatcgccta cctcatcggc aggaagagga
     1801 gtcacgcggg ctatcagacc atctcggaat tcggctccac cggctccacc ggctccaccg
     1861 gcgcggatcc accggtcgcc accatggtga gcaagggcga ggagaataac atggccatca
     1921 tcaaggagtt catgcgcttc aaggtgcgca tggagggctc cgtgaacggc cacgagttcg
     1981 agatcgaggg cgagggcgag ggccgcccct acgagggctt tcagaccgct aagctgaagg
     2041 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcat ttcacctacg
     2101 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttcaag ctgtccttcc
     2161 ccgagggctt caagtgggag cgcgtgatga actacgagga cggcggcgtg gtgaccgtga
     2221 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     2281 acttcccctc cgacggcccc gtgatgcaga agaagaccat gggctgggag gcctcctccg
     2341 agcggatgta ccccgaggac ggtgccctga agggcaagat caagatgagg ctgaagctga
     2401 aggacggcgg ccactacacc tccgaggtca agaccaccta caaggccaag aagcccgtgc
     2461 agctgcccgg cgcctacatc gtcgacatca agttggacat cacctcccac aacgaggact
     2521 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     2581 agctgtacaa gtaagcggcc gcgactctag atcataatca gccataccac atttgtagag
     2641 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     2701 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2761 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     2821 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     2881 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     2941 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     3001 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3061 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     3121 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     3181 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3241 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3301 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3361 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3421 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3481 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3541 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3601 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3661 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3721 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3781 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     3841 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     3901 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     3961 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     4021 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4081 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4141 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4201 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4261 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4321 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     4381 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4441 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4501 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4561 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4621 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4681 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4741 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     4801 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     4861 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     4921 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     4981 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     5041 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     5101 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5161 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5221 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5281 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5341 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5401 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5461 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5521 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5581 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5641 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5701 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5761 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5821 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5881 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata
     5941 gtaatcaatt a
//
