LOCUS       sequence-215601-        5952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Lysosome Membrane, Excitation: 587, Emission: 610.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5952)
  TITLE     Davidson lab - mCherry constructs
REFERENCE   2  (bases 1 to 5952)
  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"
COMMENT     Sequence Label: mCherry-Lysosomes-20
FEATURES             Location/Qualifiers
     source          1..5952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      560..569
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      564..573
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(566..628,629..1786)
                     /codon_start=1
                     /product="rat lysosome-associated membrane glycoprotein 1"
                     /label=LAMP1
                     /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF
                     LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT
                     RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI
                     VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT
                     CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG
                     MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL
                     ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR
                     SHAGYQTI"
     regulatory      1841..1850
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1847..2557
                     /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(1992..2010)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2260..2279
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2680..2801
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2717..2736)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2771..2790
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2808..3263)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2945..2966)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3157..3176
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3290..3394
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3396..3753
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3396..3587
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3419..3439)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3604..3739
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3788..4582
                     /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             3788..4582
                     /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(3842..3861)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4452..4471
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4770..4789)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4814..4861
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5190..5778
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5679..5698
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagcg ccaccatggc ggccccgggc gcccggcggc cgctgctcct
      601 gttgctgctg gcaggccttg cacacagcgc cccagcactg ttcgaggtga aagacaacaa
      661 cggcacagcg tgtataatgg ccagcttctc tgcctccttt ctgaccacct atgaggctgg
      721 acatgtttct aaggtctcga atatgaccct gccagcctct gcagaagtcc tgaagaatag
      781 cagctcttgt ggtgaaaaga atgcttctga gcccaccctc gcaatcacct ttggagaagg
      841 atatttactg aaactcacct tcacaaaaaa cacaacacgt tacagtgtcc agcacatgta
      901 tttcacatat aacctgtcag acacacaatt ctttcccaat gccagctcca aagggcccga
      961 cactgtggat tccacaactg acatcaaggc agacatcaac aaaacatacc gatgtgtcag
     1021 cgacatcagg gtctacatga agaatgtgac cattgtgctc tgggacgcta ctatccaggc
     1081 ctacctgccg agtagcaact tcagcaagga agagacacgc tgcccacagg atcaaccttc
     1141 cccaactact gggccaccca gcccctcacc accacttgtg cccacaaacc ccagtgtgtc
     1201 caagtacaat gtgactggtg acaatggaac ctgcctgctg gcctctatgg cactgcaact
     1261 caacatcacc tacatgaaga aggacaacac gactgtgacc agagcattca acatcaaccc
     1321 aagtgacaaa tatagtggga cttgcggtgc ccagttggtg accctgaagg tggggaacaa
     1381 gagcagagtc ctggagctgc agtttgggat gaatgccact tctagcctgt ttttcctgca
     1441 aggagttcag ttgaacatga ctcttcctga tgccatagag cccacgttca gcacctccaa
     1501 ctattccctg aaagctcttc aggccagtgt cggcaactca tacaagtgca acagtgagga
     1561 gcacatcttt gtcagcaagg cgctcgccct caatgtcttc agcgtgcaag tccaggcttt
     1621 cagggtagaa agtgacaggt ttgggtctgt ggaagagtgt gtacaggacg gtaacaacat
     1681 gctgatcccc attgctgtgg gcggggccct ggcagggctg gtcctcatcg tcctcatcgc
     1741 ctacctcatc ggcaggaaga ggagtcacgc gggctatcag accatctcgg aattcggctc
     1801 caccggctcc accggctcca ccggcgcgga tccaccggtc gccaccatgg tgagcaaggg
     1861 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     1921 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     1981 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     2041 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     2101 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     2161 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     2221 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     2281 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     2341 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     2401 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     2461 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     2521 ccactccacc ggcggcatgg acgagctgta caagtaaagc ggccgcgact ctagatcata
     2581 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2641 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2701 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     2761 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     2821 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     2881 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     2941 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3001 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     3061 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     3121 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     3181 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3241 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3301 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3361 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     3421 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3481 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3541 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3601 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3661 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3721 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     3781 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3841 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3901 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3961 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4021 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4081 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4141 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4201 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4261 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     4321 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4381 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4441 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4501 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4561 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4621 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4681 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4741 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     4801 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     4861 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     4921 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     4981 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     5041 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5101 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5161 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5221 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5281 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5341 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5401 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5461 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5521 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5581 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5641 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5701 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5761 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5821 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5881 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     5941 tatggagttc cg
//
