LOCUS       sequence-461117-        5961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Visualization of lysosome in mammalian cells using mGold2s.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5961)
  AUTHORS   Lee J, Lai S, Yang S, Zhao S, Blanco FA, Lyons AC, Merino-Urteaga R,
            Ahrens JF, Nguyen NA, Liu H, Liu Z, Lambert GG, Shaner NC, Chen L, 
            Tolias KF, Zhang J, Ha T, St-Pierre F
  TITLE     Bright and photostable yellow fluorescent proteins for extended 
            imaging.
  JOURNAL   Nat Commun. 2025 Apr 4;16(1):3241. doi: 10.1038/s41467-025-58223-5.
  PUBMED    40185748
REFERENCE   2  (bases 1 to 5961)
  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 
            Commun."; date: "2025-04-4"; volume: "16(1):3241. doi"; pages: " 
            10.1038/s41467-025-58223-5"
COMMENT     Sequence Label: pCMV-mGold2s-Lysosome-N-20
FEATURES             Location/Qualifiers
     source          1..5961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        78..381
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..585
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     536..556
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      637..646
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      641..650
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(643..705,706..1863)
                     /codon_start=1
                     /product="rat lysosome-associated membrane glycoprotein 1"
                     /label=LAMP1
                     /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF
                     LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT
                     RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI
                     VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT
                     CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG
                     MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL
                     ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR
                     SHAGYQTI"
     regulatory      1918..1927
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1924..1926,1927..1929,1930..2643)
                     /codon_start=1
                     /product="photostable derivative of mVenus (Lee et al., 
                     2020)"
                     /label=mGold
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDINGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTSLGYGLQCFARYPDNMKRHDFFKSAMPEGYVQERTIFFEDD
                     GNYKTRAEVKFEGGTLVNRVELKGIDFKEDGNILGHKLEYNYNCHNVFITADRQQNGIK
                     VNFKIRHNIEVGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLSMDELYK"
     primer_bind     complement(2230..2249)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2577..2598
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2766..2887
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2803..2822)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2857..2876
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2894..3349)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3031..3052)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3243..3262
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3376..3480
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3482..3839
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3482..3673
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3505..3525)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3690..3825
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3874..4668
                     /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             3874..4668
                     /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(3928..3947)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4538..4557
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4856..4875)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4900..4947
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5276..5864
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5765..5784
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
       61 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      121 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      181 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      241 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      301 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      361 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      421 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      481 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      541 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
      601 cagatccgct agcgctaccg gactcagatc tcgagcgcca ccatggcggc cccgggcgcc
      661 cggcgtccgc tgctcctgtt gctgctggca ggccttgcac acagcgcccc agcactgttc
      721 gaggtgaaag acaacaacgg cacagcgtgt ataatggcca gcttctctgc ctcctttctg
      781 accacctatg aggctggaca tgtttctaag gtctcgaata tgaccctgcc agcctctgca
      841 gaagtcctga agaatagcag ctcttgtggt gaaaagaatg cttctgagcc caccctcgca
      901 atcacctttg gagaaggata tttactgaaa ctcaccttca caaaaaacac aacacgttac
      961 agtgtccagc acatgtattt cacatataac ctgtcagaca cacaattctt tcccaatgcc
     1021 agctccaaag ggcccgacac tgtggattcc acaactgaca tcaaggcaga catcaacaaa
     1081 acataccgat gtgtcagcga catcagggtc tacatgaaga atgtgaccat tgtgctctgg
     1141 gacgctacta tccaggccta cctgccgagt agcaacttca gcaaggaaga gacacgctgc
     1201 ccacaggatc aaccttcccc aactactggg ccacccagcc cctcaccacc acttgtgccc
     1261 acaaacccca gtgtgtccaa gtacaatgtg actggtgaca atggaacctg cctgctggcc
     1321 tctatggcac tgcaactcaa catcacctac atgaagaagg acaacacgac tgtgaccaga
     1381 gcattcaaca tcaacccaag tgacaaatat agtgggactt gcggtgccca gttggtgacc
     1441 ctgaaggtgg ggaacaagag cagagtcctg gagctgcagt ttgggatgaa tgccacttct
     1501 agcctgtttt tcctgcaagg agttcagttg aacatgactc ttcctgatgc catagagccc
     1561 acgttcagca cctccaacta ttccctgaaa gctcttcagg ccagtgtcgg caactcatac
     1621 aagtgcaaca gtgaggagca catctttgtc agcaaggcgc tcgccctcaa tgtcttcagc
     1681 gtgcaagtcc aggctttcag ggtagaaagt gacaggtttg ggtctgtgga agagtgtgta
     1741 caggacggta acaacatgct gatccccatt gctgtgggcg gggccctggc agggctggtc
     1801 ctcatcgtcc tcatcgccta cctcatcggc aggaagagga gtcacgcggg ctatcagacc
     1861 atctcggaat tcggctccac cggctccacc ggctccaccg gcgcggatcc accggtcgcc
     1921 accatggcga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1981 gacggcgaca taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     2041 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     2101 accctcgtga ccagcctggg ctacggcctg cagtgcttcg cccgctaccc cgacaacatg
     2161 aagcggcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2221 ttcttcgagg acgacggcaa ctacaagacc cgcgccgagg tgaagtttga gggcggcacc
     2281 ctggtgaacc gcgtcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2341 cacaagctgg agtacaacta caactgccac aacgtcttta tcaccgccga caggcagcag
     2401 aacggcatca aggtcaactt caagatccgc cacaacatcg aggtcggcgg cgtgcagctc
     2461 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     2521 cactacctga gctaccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     2581 gtcctgctgg agttcgtgac cgccgccggg atcactctca gcatggacga gctgtacaag
     2641 taaagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     2701 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     2761 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     2821 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     2881 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     2941 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3001 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     3061 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     3121 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     3181 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     3241 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     3301 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     3361 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3421 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3481 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3541 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3601 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3661 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3721 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     3781 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     3841 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3901 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3961 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4021 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     4081 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4141 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4201 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4261 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4321 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4381 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     4441 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4501 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4561 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4621 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4681 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4741 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4801 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     4861 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     4921 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     4981 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     5041 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     5101 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     5161 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5221 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5281 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5341 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5401 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5461 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5521 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5581 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5641 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5701 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5761 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5821 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5881 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5941 cccctgattc tgtggataac c
//
