LOCUS       sequence-430619-        5950 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 5950)
  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 5950)
  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-mScarlet-I
FEATURES             Location/Qualifiers
     source          1..5950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     42..62
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(168..251,252..1418)
                     /codon_start=1
                     /product="human lysosome-associated membrane glycoprotein 
                     1"
                     /label=LAMP1
                     /translation="MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMAN
                     FSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNF
                     TRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHM
                     NNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKY
                     NVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSE
                     GTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNA
                     EEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLIV
                     LIAYLVGRKRSHAGYQTI"
     regulatory      1434..1443
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1440..2135
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1573..1591)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1841..1860
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2261..2382
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2298..2317)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2352..2371
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2389..2844)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2526..2547)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2738..2757
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2871..2975
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2977..3334
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2977..3168
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3000..3020)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3185..3320
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3369..4163
                     /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             3369..4163
                     /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(3423..3442)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4033..4052
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4351..4370)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4395..4442
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4771..5359
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5260..5279
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5534..5837
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(5838..5950,1..91)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
       61 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg
      121 ctaccggact cagatctcga gctcaagctt cgaattctgc agtcgacatg gcggcccccg
      181 gcagcgcccg gcgacccctg ctgctgctac tgctgttgct gctgctcggc ctcatgcatt
      241 gtgcgtcagc agcaatgttt atggtgaaaa atggcaacgg gaccgcgtgc ataatggcca
      301 acttctctgc tgccttctca gtgaactacg acaccaagag tggccctaag aacatgacct
      361 ttgacctgcc atcagatgcc acagtggtgc tcaaccgcag ctcctgtgga aaagagaaca
      421 cttctgaccc cagtctcgtg attgcttttg gaagaggaca tacactcact ctcaatttca
      481 cgagaaatgc aacacgttac agcgtccagc tcatgagttt tgtttataac ttgtcagaca
      541 cacacctttt ccccaatgcg agctccaaag aaatcaagac tgtggaatct ataactgaca
      601 tcagggcaga tatagataaa aaatacagat gtgttagtgg cacccaggtc cacatgaaca
      661 acgtgaccgt aacgctccat gatgccacca tccaggcgta cctttccaac agcagcttca
      721 gcaggggaga gacacgctgt gaacaagaca ggccttcccc aaccacagcg ccccctgcgc
      781 cacccagccc ctcgccctca cccgtgccca agagcccctc tgtggacaag tacaacgtga
      841 gcggcaccaa cgggacctgc ctgctggcca gcatggggct gcagctgaac ctcacctatg
      901 agaggaagga caacacgacg gtgacaaggc ttctcaacat caaccccaac aagacctcgg
      961 ccagcgggag ctgcggcgcc cacctggtga ctctggagct gcacagcgag ggcaccaccg
     1021 tcctgctctt ccagttcggg atgaatgcaa gttctagccg gtttttccta caaggaatcc
     1081 agttgaatac aattcttcct gacgccagag accccgcctt taaagctgcc aacggctccc
     1141 tgcgagcgct gcaggccaca gtcggcaatt cctacaagtg caacgcggag gagcacgtcc
     1201 gtgtcacgaa ggcgttttca gtcaatatat tcaaagtgtg ggtccaggct ttcaaggtgg
     1261 aaggtggcca gtttggctct gtggaggagt gtctgctgga cgagaacagc atgctgatcc
     1321 ccatcgctgt gggtggtgcc ctggcggggc tggtcctcat cgtcctcatc gcctacctcg
     1381 tcggcaggaa gaggagtcac gcaggctacc agactatcaa ggatccaccg gtcgccacca
     1441 tggtgagcaa gggcgaggca gtgatcaagg agttcatgcg gttcaaggtg cacatggagg
     1501 gctccatgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     1561 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc tcctgggaca
     1621 tcctgtcccc tcagttcatg tacggctcca gggccttcat caagcacccc gccgacatcc
     1681 ccgactacta taagcagtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     1741 aggacggcgg cgccgtgacc gtgacccagg acacctccct ggaggacggc accctgatct
     1801 acaaggtgaa gctccgcggc accaacttcc ctcctgacgg ccccgtaatg cagaagaaga
     1861 caatgggctg ggaagcgtcc accgagcggt tgtaccccga ggacggcgtg ctgaagggcg
     1921 acattaagat ggccctgcgc ctgaaggacg gcggccgcta cctggcggac ttcaagacca
     1981 cctacaaggc caagaagccc gtgcagatgc ccggcgccta caacgtcgac cgcaagttgg
     2041 acatcacctc ccacaacgag gactacaccg tggtggaaca gtacgaacgc tccgagggcc
     2101 gccactccac cggcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat
     2161 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2221 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2281 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2341 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     2401 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2461 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2521 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2581 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2641 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2701 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     2761 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     2821 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2881 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2941 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     3001 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3061 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3121 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3181 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3241 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3301 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     3361 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3421 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3481 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3541 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3601 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3661 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3721 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3781 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3841 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     3901 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3961 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4021 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4081 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4141 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4201 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4261 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4321 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     4381 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     4441 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     4501 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     4561 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     4621 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4681 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4741 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4801 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4861 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4921 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4981 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5041 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5101 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5161 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5221 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5281 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5341 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5401 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5461 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     5521 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     5581 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     5641 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     5701 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     5761 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     5821 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     5881 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     5941 accaaaatca
//
