LOCUS       sequence-190174-        5971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo visualization of lysosomes (can be used for colocalization 
            studies).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5971)
  AUTHORS   Chertkova AO, Mastop M, Postma M, van Bommel N, van der Niet S, 
            Batenburg KL, Joosen L, Gadella TWJ, Okada Y, Goedhart J
  TITLE     Robust and Bright Genetically Encoded Fluorescent Markers for 
            Highlighting Structures and Compartments in Mammalian Cells
  JOURNAL   bioRxiv (2020) 160374; doi: https://doi.org/10.1101/160374
REFERENCE   2  (bases 1 to 5971)
  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: "bioRxiv 
            (2020) 160374; doi: https://doi.org/10.1101/160374"
COMMENT     Sequence Label: Lamp1-mTurquoise2
FEATURES             Location/Qualifiers
     source          1..5971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..259
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     216..236
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(261..365)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      392..847
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(479..498)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     689..710
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    854..975
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(865..884)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     919..938
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(1098..1811,1812..1814,1815..1817))
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1143..1164)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1492..1511
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1751..1772
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1814..1823
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(join(1839..3005,3006..3089))
                     /codon_start=1
                     /product="human lysosome-associated membrane glycoprotein 
                     1"
                     /label=LAMP1
                     /translation="MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMAN
                     FSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNF
                     TRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHM
                     NNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKY
                     NVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSE
                     GTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNA
                     EEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLIV
                     LIAYLVGRKRSHAGYQTI"
     promoter        complement(3166..3369)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3195..3215)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3370..3673
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3848..4436)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3928..3947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4765..4812
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4837..4856
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5044..5838)
                     /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             complement(5044..5838)
                     /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(5155..5174)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5765..5784
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5873..5971,1..259))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5887..5971,1..51)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
       61 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
      121 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
      181 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
      241 ctggttcttt ccgcctcagg actcttcctt tttcaatatt attgaagcat ttatcagggt
      301 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
      361 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
      421 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
      481 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
      541 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
      601 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
      661 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
      721 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
      781 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
      841 acaatttacg ccttaagata cattgatgag tttggacaaa ccacaactag aatgcagtga
      901 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
      961 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     1021 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat
     1081 ctagagtcgc ggccgcttta cttgtacagc tcgtccatgc cgagagtgat cccggcggcg
     1141 gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagcttg
     1201 gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg
     1261 ttctgctggt agtggtcggc gagctgcacg ccgccgtcct cgatgttgtg gcggatcttg
     1321 aagttggcct tgatgccgtt cttctgcttg tcggcggtga tatagacgtt gtcgctaaag
     1381 tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc
     1441 agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag
     1501 ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg catggcggac
     1561 ttgaagaagt cgtgctgctt catgtggtcg gggtagcggg cgaagcactg cacgccccag
     1621 gacagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatgaac
     1681 ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac gctgaacttg
     1741 tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc
     1801 tcgcccttgc tcaccatggt ggcgaccggt ggatccttga tagtctggta gcctgcgtga
     1861 ctcctcttcc tgccgacgag gtaggcgatg aggacgatga ggaccagccc cgccagggca
     1921 ccacccacag cgatggggat cagcatgctg ttctcgtcca gcagacactc ctccacagag
     1981 ccaaactggc caccttccac cttgaaagcc tggacccaca ctttgaatat attgactgaa
     2041 aacgccttcg tgacacggac gtgctcctcc gcgttgcact tgtaggaatt gccgactgtg
     2101 gcctgcagcg ctcgcaggga gccgttggca gctttaaagg cggggtctct ggcgtcagga
     2161 agaattgtat tcaactggat tccttgtagg aaaaaccggc tagaacttgc attcatcccg
     2221 aactggaaga gcaggacggt ggtgccctcg ctgtgcagct ccagagtcac caggtgggcg
     2281 ccgcagctcc cgctggccga ggtcttgttg gggttgatgt tgagaagcct tgtcaccgtc
     2341 gtgttgtcct tcctctcata ggtgaggttc agctgcagcc ccatgctggc cagcaggcag
     2401 gtcccgttgg tgccgctcac gttgtacttg tccacagagg ggctcttggg cacgggtgag
     2461 ggcgaggggc tgggtggcgc agggggcgct gtggttgggg aaggcctgtc ttgttcacag
     2521 cgtgtctctc ccctgctgaa gctgctgttg gaaaggtacg cctggatggt ggcatcatgg
     2581 agcgttacgg tcacgttgtt catgtggacc tgggtgccac taacacatct gtatttttta
     2641 tctatatctg ccctgatgtc agttatagat tccacagtct tgatttcttt ggagctcgca
     2701 ttggggaaaa ggtgtgtgtc tgacaagtta taaacaaaac tcatgagctg gacgctgtaa
     2761 cgtgttgcat ttctcgtgaa attgagagtg agtgtatgtc ctcttccaaa agcaatcacg
     2821 agactggggt cagaagtgtt ctcttttcca caggagctgc ggttgagcac cactgtggca
     2881 tctgatggca ggtcaaaggt catgttctta gggccactct tggtgtcgta gttcactgag
     2941 aaggcagcag agaagttggc cattatgcac gcggtcccgt tgccattttt caccataaac
     3001 attgctgctg acgcacaatg catgaggccg agcagcagca acagcagtag cagcagcagg
     3061 ggtcgccggg cgctgccggg ggccgccatg tcgactgcag aattcgaagc ttgagctcga
     3121 gatctgagtc cggtagcgct agcggatctg acggttcact aaaccagctc tgcttatata
     3181 gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca
     3241 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
     3301 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa
     3361 ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     3421 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     3481 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     3541 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     3601 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
     3661 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
     3721 ctattaataa ctaatgcatg gcggtaatac ggttatccac agaatcaggg gataacgcag
     3781 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3841 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3901 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3961 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4021 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4081 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4141 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4201 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4261 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4321 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4381 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4441 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4501 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4561 gttttaaatc aatctaaagt atatatgagt aacctgaggc tatggcaggg cctgccgccc
     4621 cgacgttggc tgcgagccct gggccttcac ccgaacttgg ggggtggggt ggggaaaagg
     4681 aagaaacgcg ggcgtattgg ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc
     4741 cctgggaccg aaccccgcgt ttatgaacaa acgacccaac accgtgcgtt ttattctgtc
     4801 tttttattgc cgtcatagcg cgggttcctt ccggtattgt ctccttccgt gtttcagtta
     4861 gcctccccct agggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc
     4921 agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat
     4981 cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt catttcgaac cccagagtcc
     5041 cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc
     5101 gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc
     5161 acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat
     5221 gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt
     5281 cacgacgaga tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca gttcggctgg
     5341 cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg
     5401 agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc
     5461 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
     5521 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
     5581 ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag
     5641 ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
     5701 aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
     5761 ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa
     5821 tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa
     5881 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
     5941 cctcggcctc tgcataaata aaaaaaatta g
//
