LOCUS       sequence-372787-        5615 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Lysosome Membrane, Excitation: 568, Emission: 592.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5615)
  TITLE     Davidson lab - mApple constructs
REFERENCE   2  (bases 1 to 5615)
  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: mApple-LAMP1-phLuorin-N-8
FEATURES             Location/Qualifiers
     source          1..5615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      627..636
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             633..722
                     /codon_start=1
                     /product="signal sequence from bovine prolactin"
                     /label=prolactin signal sequence
                     /translation="MDSKGSSQKGSRLLLLLVVSNLLLCQGVVS"
     CDS             723..1433
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Ng et al., 2002)"
                     /label=superecliptic pHluorin
                     /note="excited strongly by blue light at neutral pH but 
                     minimally fluorescent at acidic pH"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(750..778)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1373..1393
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     regulatory      1572..1581
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1578..2288
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF
                     EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK
                     SEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     1991..2010
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2410..2531
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2447..2466)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2501..2520
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2538..2993)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2675..2696)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2887..2906
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3020..3124
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3126..3483
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3126..3317
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3149..3169)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3334..3469
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3518..4312
                     /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             3518..4312
                     /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(3572..3591)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4182..4201
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4500..4519)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4544..4591
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4920..5508
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5409..5428
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gactcagatc tcgagcgcca ccatggacag caaaggttcg tcgcagaaag
      661 ggtcccgcct gctcctgctg ctggtggtgt caaatctact cttgtgccag ggtgtggtct
      721 ccagtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg
      781 atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa
      841 aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg ccaacacttg
      901 tcactacttt aacttatggt gttcaatgct tttcaagata cccagatcat atgaaacggc
      961 atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca
     1021 aagatgacgg gaactacaag acacgtgctg aagtcaagtt tgaaggtgat acccttgtta
     1081 atagaatcga gttaaaaggt attgatttta aagaagatgg aaacattctt ggacacaaat
     1141 tggaatacaa ctataacgat caccaggtgt acatcatggc agacaaacaa aagaatggaa
     1201 tcaaagctaa cttcaaaatt agacacaaca ttgaagatgg aggcgttcaa ctagcagacc
     1261 attatcaaca aaatactcca attggcgatg ggcccgtcct tttaccagac aaccattacc
     1321 tgtttacaac ttctactctt tcgaaagatc ccaacgaaaa gagagaccac atggtccttc
     1381 ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac aaaggatcca
     1441 acaacatgtt gatccccatt gctgtgggcg gtgccctggc agggctggtc ctcatcgtcc
     1501 tcattgccta cctcattggc aggaagagga gtcacgccgg ctatcagacc atctcagggt
     1561 ctggaccggt cgccaccatg gtgagcaagg gcgaggagaa taacatggcc atcatcaagg
     1621 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
     1681 agggcgaggg cgagggccgc ccctacgagg cctttcagac cgctaagctg aaggtgacca
     1741 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
     1801 aggtctacat taagcaccca gccgacatcc ccgactactt caagctgtcc ttccccgagg
     1861 gcttcaggtg ggagcgcgtg atgaacttcg aggacggcgg cattattcac gttaaccagg
     1921 actcctccct gcaggacggc gtgttcatct acaaggtgaa gctgcgcggc accaacttcc
     1981 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc gaggagcgga
     2041 tgtaccccga ggacggcgcc ctgaagagcg agatcaagaa gaggctgaag ctgaaggacg
     2101 gcggccacta cgccgccgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
     2161 ccggcgccta catcgtcgac atcaagttgg acatcgtgtc ccacaacgag gactacacca
     2221 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
     2281 acaagtaagc ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta
     2341 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     2401 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2461 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2521 aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     2581 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     2641 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     2701 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     2761 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     2821 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     2881 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     2941 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     3001 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3061 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3121 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3181 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3241 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3301 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3361 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3421 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3481 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3541 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3601 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3661 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     3721 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3781 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3841 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3901 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     3961 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4021 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     4081 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4141 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4201 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4261 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4321 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4381 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4441 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     4501 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     4561 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     4621 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     4681 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     4741 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     4801 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4861 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4921 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4981 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5041 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5101 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5161 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5221 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5281 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5341 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5401 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5461 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5521 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5581 ttatcccctg attctgtgga taaccgtatt accgc
//
