LOCUS       sequence-254063-        6275 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Lysosome Membrane, Excitation: 448 / 473, Emission: 
            500 / 528.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6275)
  AUTHORS   Shu X, Lev-Ram V, Deerinck TJ, Qi Y, Ramko EB, Davidson MW, Jin Y, 
            Ellisman MH, Tsien RY
  TITLE     A genetically encoded tag for correlated light and electron 
            microscopy of intact cells, tissues, and organisms.
  JOURNAL   PLoS Biol. 2011 Apr;9(4):e1001041. doi: 
            10.1371/journal.pbio.1001041. Epub 2011 Apr 5.
  PUBMED    21483721
REFERENCE   2  (bases 1 to 6275)
  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: "PLoS 
            Biol."; date: "2011-04"; volume: "9(4):e1001041. doi"; pages: " 
            10.1371/journal.pbio.1001041. Epub 2011 Apr 5"
COMMENT     Sequence Label: PA-mCherry-miniSOG-Lysosomes-20
FEATURES             Location/Qualifiers
     source          1..6275
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        125..428
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        429..632
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     583..603
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      684..693
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      688..697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(690..752,753..1910)
                     /codon_start=1
                     /product="rat lysosome-associated membrane glycoprotein 1"
                     /label=LAMP1
                     /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF
                     LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT
                     RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI
                     VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT
                     CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG
                     MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL
                     ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR
                     SHAGYQTI"
     regulatory      1965..1974
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1971..2678
                     /codon_start=1
                     /product="photoactivatable monomeric derivative of DsRed 
                     fluorescent protein (Subach et al., 2009)"
                     /label=PAmCherry1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHVFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFTWDILSPQFMYGSNAYVKHPADIPDYFKLSFPEGFKWERVMKF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEALSERMYPEDGALK
                     GEVKPRVKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNRKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2116..2134)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             2685..3005
                     /codon_start=1
                     /product="mini singlet oxygen generator, a fluorescent 
                     flavoprotein suitable for correlated light and electron 
                     microscopy (Shu et al., 2011)"
                     /label=miniSOG
                     /translation="MEKSFVITDPRLPDNPIIFASDGFLELTEYSREEILGRNGRFLQG
                     PETDQATVQKIRDAIRDQREITVQLINYTKSGKKFWNLLHLQPMRDQKGELQYFIGVQL
                     DG"
     polyA_signal    3127..3248
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3164..3183)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3218..3237
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3255..3710)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3392..3413)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3604..3623
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3737..3841
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3843..4200
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3843..4034
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3866..3886)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4051..4186
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4235..5029
                     /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             4235..5029
                     /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(4289..4308)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4899..4918
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5217..5236)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5261..5308
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5637..6225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6126..6145
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
       61 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
      121 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      181 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      241 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      301 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      361 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      421 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      481 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      541 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      601 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc
      661 gctaccggac tcagatctcg agcgccacca tggcggcccc gggcgcccgg cgtccgctgc
      721 tcctgttgct gctggcaggc cttgcacaca gcgccccagc actgttcgag gtgaaagaca
      781 acaacggcac agcgtgtata atggccagct tctctgcctc ctttctgacc acctatgagg
      841 ctggacatgt ttctaaggtc tcgaatatga ccctgccagc ctctgcagaa gtcctgaaga
      901 atagcagctc ttgtggtgaa aagaatgctt ctgagcccac cctcgcaatc acctttggag
      961 aaggatattt actgaaactc accttcacaa aaaacacaac acgttacagt gtccagcaca
     1021 tgtatttcac atataacctg tcagacacac aattctttcc caatgccagc tccaaagggc
     1081 ccgacactgt ggattccaca actgacatca aggcagacat caacaaaaca taccgatgtg
     1141 tcagcgacat cagggtctac atgaagaatg tgaccattgt gctctgggac gctactatcc
     1201 aggcctacct gccgagtagc aacttcagca aggaagagac acgctgccca caggatcaac
     1261 cttccccaac tactgggcca cccagcccct caccaccact tgtgcccaca aaccccagtg
     1321 tgtccaagta caatgtgact ggtgacaatg gaacctgcct gctggcctct atggcactgc
     1381 aactcaacat cacctacatg aagaaggaca acacgactgt gaccagagca ttcaacatca
     1441 acccaagtga caaatatagt gggacttgcg gtgcccagtt ggtgaccctg aaggtgggga
     1501 acaagagcag agtcctggag ctgcagtttg ggatgaatgc cacttctagc ctgtttttcc
     1561 tgcaaggagt tcagttgaac atgactcttc ctgatgccat agagcccacg ttcagcacct
     1621 ccaactattc cctgaaagct cttcaggcca gtgtcggcaa ctcatacaag tgcaacagtg
     1681 aggagcacat ctttgtcagc aaggcgctcg ccctcaatgt cttcagcgtg caagtccagg
     1741 ctttcagggt agaaagtgac aggtttgggt ctgtggaaga gtgtgtacag gacggtaaca
     1801 acatgctgat ccccattgct gtgggcgggg ccctggcagg gctggtcctc atcgtcctca
     1861 tcgcctacct catcggcagg aagaggagtc acgcgggcta tcagaccatc tcggaattcg
     1921 gctccaccgg ctccaccggc tccaccggcg cggatccacc ggtcgccacc atggtgagca
     1981 agggcgagga ggataacatg gccatcatta aggagttcat gcgcttcaag gtgcacatgg
     2041 aggggtccgt gaacggccac gtgttcgaga tcgagggcga gggcgagggc cgcccctacg
     2101 agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcacctggg
     2161 acatcctgag ccctcagttc atgtacggct ccaatgccta cgtgaagcac cccgccgaca
     2221 tccccgacta ctttaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaaat
     2281 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca
     2341 tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgtg atgcagaaga
     2401 agaccatggg ctgggaggcc ctctccgagc ggatgtaccc cgaggacggc gccctgaagg
     2461 gcgaggtcaa gcccagagtg aagctgaagg acggcggcca ctacgacgct gaggtcaaga
     2521 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aaccgcaagc
     2581 tggacatcac cagccacaac gaggactaca ccatcgtgga gcagtacgag agagccgagg
     2641 gccgccactc caccggcggc atggacgagc tgtacaaggg cggcatggag aagagcttcg
     2701 tgatcaccga cccccgcctg cccgacaacc ccatcatctt cgccagcgac ggcttcctgg
     2761 agctgaccga gtacagcaga gaggagatcc tgggccgcaa cggccgcttc ctgcagggcc
     2821 ccgagaccga ccaggccacc gtgcagaaga tccgcgacgc catccgcgac cagcgcgaga
     2881 tcaccgtgca gctgatcaac tacaccaaga gcggcaagaa gttctggaac ctgctgcacc
     2941 tgcagcccat gcgcgaccag aagggcgagc tgcagtactt catcggcgtg cagctggacg
     3001 gctaagcggc cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt
     3061 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3121 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3181 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3241 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3301 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3361 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3421 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     3481 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     3541 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3601 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     3661 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     3721 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3781 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3841 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3901 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3961 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4021 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4081 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4141 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4201 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4261 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4321 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4381 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     4441 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4501 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4561 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4621 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4681 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4741 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4801 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4861 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4921 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4981 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5041 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5101 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5161 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     5221 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     5281 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5341 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     5401 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     5461 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     5521 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5581 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5641 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5701 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5761 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5821 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5881 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5941 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6001 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6061 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6121 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6181 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6241 gcctttttac ggttcctggc cttttgctgg ccttt
//
