LOCUS       sequence-338156-        6437 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Homo sapiens lysosomal-associated membrane protein 1 
            (LAMP1) tagged with mCherry and AIP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6437)
  AUTHORS   Miyamoto T, Rho E, Sample V, Akano H, Magari M, Ueno T, Gorshkov K, 
            Chen M, Tokumitsu H, Zhang J, Inoue T
  TITLE     Compartmentalized AMPK signaling illuminated by genetically encoded 
            molecular sensors and actuators.
  JOURNAL   Cell Rep. 2015 Apr 28;11(4):657-70. doi: 
            10.1016/j.celrep.2015.03.057. Epub 2015 Apr 16.
  PUBMED    25892241
REFERENCE   2  (bases 1 to 6437)
  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"; doi: 
            "10.1016/j.celrep.2015.03.057"; journalName: "Cell Rep"; date: 
            "2015-04-28- 28"; volume: "11"; issue: "4"; pages: "657-70"
COMMENT     Sequence Label: LAMP-mChF-AIP
FEATURES             Location/Qualifiers
     source          1..6437
                     /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      552..561
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      556..565
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(558..641,642..1808)
                     /codon_start=1
                     /product="human lysosome-associated membrane glycoprotein 
                     1"
                     /label=LAMP1
                     /translation="MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMAN
                     FSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNF
                     TRNATRYSVQLMSFAYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHM
                     NNVTVTPHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKY
                     NVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSE
                     GTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNA
                     EEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLIV
                     LIAYLVGRKRSHAGYQTI"
     CDS             1827..2534
                     /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(1972..1990)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2240..2259
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2547..2867
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    3165..3286
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3202..3221)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3256..3275
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3293..3748)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3430..3451)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3642..3661
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3775..3879
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3881..4238
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3881..4072
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3904..3924)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4089..4224
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4273..5067
                     /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             4273..5067
                     /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(4327..4346)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4937..4956
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5255..5274)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5299..5346
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5675..6263
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6164..6183
                     /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 ccggtgaatt cgccaccatg gcggcccccg gcagcgcccg gcgacccctg ctgctgctac
      601 tgctgttgct gctgctcggc ctcatgcatt gtgcgtcagc agcaatgttt atggtgaaaa
      661 atggcaacgg gaccgcgtgc ataatggcca acttctctgc tgccttctca gtgaactacg
      721 acaccaagag tggccctaag aacatgacct ttgacctgcc atcagatgcc acagtggtgc
      781 tcaaccgcag ctcctgtgga aaagagaaca cttctgaccc cagtctcgtg attgcttttg
      841 gaagaggaca tacactcact ctcaatttca cgagaaatgc aacacgttac agcgtccagc
      901 tcatgagttt tgcttataac ttgtcagaca cacacctttt ccccaatgcg agctccaaag
      961 aaatcaagac tgtggaatct ataactgaca tcagggcaga tatagataaa aaatacagat
     1021 gtgttagtgg cacccaggtc cacatgaaca acgtgaccgt aacgccccat gatgccacca
     1081 tccaggcgta cctttccaac agcagcttca gcaggggaga gacacgctgt gaacaagaca
     1141 ggccttcccc aaccacagcg ccccctgcgc cacccagccc ctcgccctca cccgtgccca
     1201 agagcccctc tgtggacaag tacaacgtga gcggcaccaa cgggacctgc ctgctggcca
     1261 gcatggggct gcagctgaac ctcacctatg agaggaagga caacacgacg gtgacaaggc
     1321 ttctcaacat caaccccaac aagacctcgg ccagcgggag ctgcggcgcc cacctggtga
     1381 ctctggagct gcacagcgag ggcaccaccg tcctgctctt ccagttcggg atgaatgcaa
     1441 gttctagccg gtttttccta caaggaatcc agttgaatac aattcttcct gacgccagag
     1501 accctgcctt taaagctgcc aacggctccc tgcgagcgct gcaggccaca gtcggcaatt
     1561 cctacaagtg caacgcggag gagcacgtcc gtgtcacgaa ggcgttttca gtcaatatat
     1621 tcaaagtgtg ggtccaggct ttcaaggtgg aaggtggcca gtttggctct gtggaggagt
     1681 gtctgctgga cgagaacagc atgctgatcc ccatcgctgt gggtggtgcc ctggcggggc
     1741 tggtcctcat cgtcctcatc gcctacctcg tcggcaggaa gaggagtcac gcaggctacc
     1801 agactatcgg atccggagca agtggaatgg tgagcaaggg cgaggaggat aacatggcca
     1861 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
     1921 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     1981 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
     2041 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
     2101 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
     2161 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     2221 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
     2281 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     2341 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     2401 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     2461 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     2521 acgaactgta caagagtgct ggtggtggag tgcaggtgga aaccatctcc ccaggagacg
     2581 ggcgcacctt ccccaagcgc ggccagacct gcgtggtgca ctacaccggg atgcttgaag
     2641 atggaaagaa atttgattcc tcccgggaca gaaacaagcc ctttaagttt atgctaggca
     2701 agcaggaggt gatccgaggc tgggaagaag gggttgccca gatgagtgtg ggtcagagag
     2761 ccaaactgac tatatctcca gattatgcct atggtgccac tgggcaccca ggcatcatcc
     2821 caccacatgc cactctcgtc ttcgatgtgg agcttctaaa actggaaagt gctggtggta
     2881 gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc tggtggtcct cgagctcaag
     2941 cttcgaattc aggatcagga gagggatcaa ccaagatgag aagagtggct actctggtgg
     3001 accttggaac cggaggatca gagctctgac tgcagggatc caccggatct agataactga
     3061 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3121 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3181 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3241 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     3301 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3361 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3421 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3481 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3541 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3601 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3661 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3721 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3781 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3841 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     3901 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3961 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4021 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4081 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4141 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4201 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4261 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4321 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4381 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4441 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4501 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4561 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4621 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4681 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4741 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4801 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4861 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4921 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4981 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5041 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5101 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5161 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5221 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     5281 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     5341 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     5401 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5461 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5521 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5581 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5641 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5701 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5761 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5821 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5881 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5941 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6001 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6061 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6121 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6181 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6241 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6301 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6361 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
     6421 ccatatatgg agttccg
//
