LOCUS       sequence-339928-        5484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for lysosomal damage.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5484)
  AUTHORS   Papadopoulos C, Kirchner P, Bug M, Grum D, Koerver L, Schulze N, 
            Poehler R, Dressler A, Fengler S, Arhzaouy K, Lux V, Ehrmann M, 
            Weihl CC, Meyer H
  TITLE     VCP/p97 cooperates with YOD1, UBXD1 and PLAA to drive clearance of 
            ruptured lysosomes by autophagy.
  JOURNAL   EMBO J. 2016 Oct 17. pii: e201695148.
  PUBMED    27753622
REFERENCE   2  (bases 1 to 5484)
  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: "EMBO J. 
            2016 Oct 17. pii: e201695148."
COMMENT     Sequence Label: pmCherry-Gal3
FEATURES             Location/Qualifiers
     source          1..5484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2..709
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(147..165)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     415..434
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1650..1771
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1687..1706)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1741..1760
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1778..2233)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1915..1936)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2127..2146
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2260..2364
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2366..2723
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2366..2557
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2389..2409)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2574..2709
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2758..3552
                     /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             2758..3552
                     /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(2812..2831)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3422..3441
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3740..3759)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3784..3831
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4160..4748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4649..4668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4934..5237
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5238..5441
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5392..5412
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5480..5484,1..5)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
       61 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
      121 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
      181 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
      241 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
      301 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
      361 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
      421 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
      481 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
      541 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
      601 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
      661 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt ccggactcag
      721 atctcgagct caagctttgg cagacaattt ttcgctccat gatgcgttat ctgggtctgg
      781 aaacccaaac cctcaaggat ggcctggcgc atgggggaac cagcctgctg gggcaggggg
      841 ctacccaggg gcttcctatc ctggggccta ccccgggcag gcacccccag gggcttatcc
      901 tggacaggca cctccaggcg cctaccctgg agcacctgga gcttatcccg gagcacctgc
      961 acctggagtc tacccagggc cacccagcgg ccctggggcc tacccatctt ctggacagcc
     1021 aagtgccacc ggagcctacc ctgccactgg cccctatggc gcccctgctg ggccactgat
     1081 tgtgccttat aacctgcctt tgcctggggg agtggtgcct cgcatgctga taacaattct
     1141 gggcacggtg aagcccaatg caaacagaat tgctttagat ttccaaagag ggaatgatgt
     1201 tgccttccac tttaacccac gcttcaatga gaacaacagg agagtcattg tttgcaatac
     1261 aaagctggat aataactggg gaagggaaga aagacagtcg gttttcccat ttgaaagtgg
     1321 gaaaccattc aaaatacaag tactggttga acctgaccac ttcaaggttg cagtgaatga
     1381 tgctcacttg ttgcagtaca atcatcgggt taaaaaactc aatgaaatca gcaaactggg
     1441 aatttctggt gacatagacc tcaccagtgc ttcatatacc atgatataag aattctgcag
     1501 tcgacggtac cgcgggcccg ggatccaccg gatctagata actgatcata atcagccata
     1561 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     1621 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     1681 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     1741 gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg
     1801 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     1861 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     1921 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     1981 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     2041 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     2101 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     2161 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     2221 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2281 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     2341 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     2401 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2461 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2521 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     2581 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     2641 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     2701 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     2761 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     2821 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     2881 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     2941 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3001 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3061 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3121 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3181 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     3241 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     3301 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     3361 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     3421 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     3481 gtgctttacg gtatcgccgc tcccgattca cagcgcatcg ccttctatcg ccttcttgac
     3541 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     3601 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     3661 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     3721 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     3781 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     3841 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     3901 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     3961 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     4021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4801 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4861 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     4921 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     4981 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     5041 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     5101 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     5161 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     5221 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     5281 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     5341 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     5401 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     5461 tccgctagcg ctaccggtcg ccac
//
