LOCUS       sequence-257391-        5954 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Lysosome Membrane, Excitation: 559, Emission: 600.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5954)
  AUTHORS   Lam AJ, St-Pierre F, Gong Y, Marshall JD, Cranfill PJ, Baird MA, 
            McKeown MR, Wiedenmann J, Davidson MW, Schnitzer MJ, Tsien RY, Lin 
            MZ
  TITLE     Improving FRET dynamic range with bright green and red fluorescent 
            proteins.
  JOURNAL   Nat Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171.
  PUBMED    22961245
REFERENCE   2  (bases 1 to 5954)
  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: "Nat 
            Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171."
COMMENT     Sequence Label: mRuby2-Lysosomes-20
FEATURES             Location/Qualifiers
     source          1..5954
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        128..431
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        432..635
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     586..606
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      687..696
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      691..700
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(693..755,756..1913)
                     /codon_start=1
                     /product="rat lysosome-associated membrane glycoprotein 1"
                     /label=LAMP1
                     /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF
                     LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT
                     RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI
                     VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT
                     CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG
                     MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL
                     ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR
                     SHAGYQTI"
     regulatory      1968..1977
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1974..2684
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     polyA_signal    2809..2930
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2846..2865)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2900..2919
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2937..3392)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3074..3095)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3286..3305
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3419..3523
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3525..3882
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3525..3716
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3548..3568)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3733..3868
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3917..4711
                     /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             3917..4711
                     /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(3971..3990)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4581..4600
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4899..4918)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4943..4990
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5319..5907
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5808..5827
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
       61 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      121 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      181 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      241 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      301 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      361 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      421 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      481 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      541 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      601 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      661 agcgctaccg gactcagatc tcgagcgcca ccatggcggc cccgggcgcc cggcgtccgc
      721 tgctcctgtt gctgctggca ggccttgcac acagcgcccc agcactgttc gaggtgaaag
      781 acaacaacgg cacagcgtgt ataatggcca gcttctctgc ctcctttctg accacctatg
      841 aggctggaca tgtttctaag gtctcgaata tgaccctgcc agcctctgca gaagtcctga
      901 agaatagcag ctcttgtggt gaaaagaatg cttctgagcc caccctcgca atcacctttg
      961 gagaaggata tttactgaaa ctcaccttca caaaaaacac aacacgttac agtgtccagc
     1021 acatgtattt cacatataac ctgtcagaca cacaattctt tcccaatgcc agctccaaag
     1081 ggcccgacac tgtggattcc acaactgaca tcaaggcaga catcaacaaa acataccgat
     1141 gtgtcagcga catcagggtc tacatgaaga atgtgaccat tgtgctctgg gacgctacta
     1201 tccaggccta cctgccgagt agcaacttca gcaaggaaga gacacgctgc ccacaggatc
     1261 aaccttcccc aactactggg ccacccagcc cctcaccacc acttgtgccc acaaacccca
     1321 gtgtgtccaa gtacaatgtg actggtgaca atggaacctg cctgctggcc tctatggcac
     1381 tgcaactcaa catcacctac atgaagaagg acaacacgac tgtgaccaga gcattcaaca
     1441 tcaacccaag tgacaaatat agtgggactt gcggtgccca gttggtgacc ctgaaggtgg
     1501 ggaacaagag cagagtcctg gagctgcagt ttgggatgaa tgccacttct agcctgtttt
     1561 tcctgcaagg agttcagttg aacatgactc ttcctgatgc catagagccc acgttcagca
     1621 cctccaacta ttccctgaaa gctcttcagg ccagtgtcgg caactcatac aagtgcaaca
     1681 gtgaggagca catctttgtc agcaaggcgc tcgccctcaa tgtcttcagc gtgcaagtcc
     1741 aggctttcag ggtagaaagt gacaggtttg ggtctgtgga agagtgtgta caggacggta
     1801 acaacatgct gatccccatt gctgtgggcg gggccctggc agggctggtc ctcatcgtcc
     1861 tcatcgccta cctcatcggc aggaagagga gtcacgcggg ctatcagacc atctcggaat
     1921 tcggctccac cggctccacc ggctccaccg gcgcggatcc accggtcgcc accatggtgt
     1981 ctaagggcga agagctgatc aaggaaaata tgcgtatgaa ggtggtcatg gaaggttcgg
     2041 tcaacggcca ccaattcaaa tgcacaggtg aaggagaagg caatccgtac atgggaactc
     2101 aaaccatgag gatcaaagtc atcgagggag gacccctgcc atttgccttt gacattcttg
     2161 ccacgtcgtt catgtatggc agccgtactt ttatcaagta cccgaaaggc attcctgatt
     2221 tctttaaaca gtcctttcct gagggtttta cttgggaaag agttacgaga tacgaagatg
     2281 gtggagtcgt caccgtcatg caggacacca gccttgagga tggctgtctc gtttaccacg
     2341 tccaagtcag aggggtaaac tttccctcca atggtcccgt gatgcagaag aagaccaagg
     2401 gttgggagcc taatacagag atgatgtatc cagcagatgg tggtctgagg ggatacactc
     2461 atatggcact gaaagttgat ggtggtggcc atctgtcttg ctctttcgta acaacttaca
     2521 ggtcaaaaaa gaccgtcggg aacatcaaga tgcccggtat ccatgccgtt gatcaccgcc
     2581 tggaaaggtt agaggaaagt gacaatgaaa tgttcgtagt acaacgcgaa cacgcagttg
     2641 ccaagttcgc cgggcttggt ggtgggatgg acgagctgta caagtaagcg gccgcgactc
     2701 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     2761 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     2821 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2881 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     2941 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3001 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3061 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     3121 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3181 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3241 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3301 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3361 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     3421 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3481 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     3541 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3601 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3661 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3721 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3781 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3841 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     3901 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3961 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4021 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4081 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4141 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4201 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4261 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4321 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4381 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4441 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4501 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4561 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4621 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4681 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4741 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4801 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4861 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     4921 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     4981 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     5041 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     5101 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     5161 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5221 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5281 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5341 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5401 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5461 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5521 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5581 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5641 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5701 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5761 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5821 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5881 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5941 gccttttgct ggcc
//
