LOCUS       sequence-261700-        5954 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Lysosome Membrane, Excitation: 399, Emission: 456.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5954)
  AUTHORS   Subach OM, Cranfill PJ, Davidson MW, Verkhusha VV
  TITLE     An enhanced monomeric blue fluorescent protein with the high 
            chemical stability of the chromophore.
  JOURNAL   PLoS One. 2011;6(12):e28674. Epub 2011 Dec 8.
  PUBMED    22174863
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: "PLoS One.";
            date: "2011"; volume: "6(12)"; pages: "e28674. Epub 2011 Dec 8"
COMMENT     Sequence Label: mTagBFP2-Lysosomes-20
FEATURES             Location/Qualifiers
     source          1..5954
                     /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="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     polyA_signal    2719..2840
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2756..2775)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2810..2829
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2847..3302)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2984..3005)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3196..3215
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3329..3433
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3435..3792
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3435..3626
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3458..3478)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3643..3778
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3827..4621
                     /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             3827..4621
                     /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(3881..3900)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4491..4510
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4809..4828)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4853..4900
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5229..5817
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5718..5737
                     /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 cggcgtccgc 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 accatggtgt ctaagggcga agagctgatt aaggagaaca
     1921 tgcacatgaa gctgtacatg gagggcaccg tggacaacca tcacttcaag tgcacatccg
     1981 agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggtg gtcgagggcg
     2041 gccctctccc cttcgccttc gacatcctgg ctactagctt cctctacggc agcaagacct
     2101 tcatcaacca cacccagggc atccccgact tcttcaagca gtccttccct gagggcttca
     2161 catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca
     2221 gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcacatcca
     2281 acggccctgt gatgcagaag aaaacactcg gctgggaggc cttcaccgag acgctgtacc
     2341 ccgctgacgg cggcctggaa ggcagaaacg acatggccct gaagctcgtg ggcgggagcc
     2401 atctgatcgc aaacgccaag accacatata gatccaagaa acccgctaag aacctcaaga
     2461 tgcctggcgt ctactatgtg gactacagac tggaaagaat caaggaggcc aacaacgaga
     2521 cctacgtcga gcagcacgag gtggcagtgg ccagatactg cgacctccct agcaaactgg
     2581 ggcacaagct taattaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     2641 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     2701 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     2761 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     2821 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     2881 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     2941 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     3001 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     3061 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     3121 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     3181 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     3241 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3301 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3361 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3421 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3481 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3541 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3601 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3661 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3721 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3781 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3841 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3901 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3961 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     4021 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4081 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4141 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4201 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4261 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4321 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     4381 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4441 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4501 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4561 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4621 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4681 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4741 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     4801 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     4861 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     4921 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     4981 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     5041 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     5101 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5161 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5221 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5281 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5341 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5401 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5461 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5521 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5581 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5641 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5701 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5761 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5821 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5881 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta
     5941 atagtaatca atta
//
