LOCUS       sequence-415451-        5993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dimerization dependent fluorescent protein RA anchored to cytosolic 
            face of the lysosomal membrane with human LAMP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5993)
  AUTHORS   Miner GE, Smith SY, Showalter WK, So CM, Ragusa JV, Powers AE, 
            Zanellati MC, Hsu CH, Marchan MF, Cohen S
  TITLE     Contact-FP: A Dimerization-Dependent Fluorescent Protein Toolkit for
            Visualizing Membrane Contact Site Dynamics.
  JOURNAL   Contact (Thousand Oaks). 2024 Feb 4;7:25152564241228911. doi: 
            10.1177/25152564241228911. eCollection 2024 Jan-Dec.
  PUBMED    38327561
REFERENCE   2  (bases 1 to 5993)
  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: "Contact 
            (Thousand Oaks)."; date: "2024-02-4"; volume: "7:25152564241228911. 
            doi"; pages: " 10.1177/25152564241228911. eCollection 2024 Jan-Dec"
COMMENT     Sequence Label: Lyso-RA
FEATURES             Location/Qualifiers
     source          1..5993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        320..523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     474..494
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      557..566
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      561..570
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(563..646,647..1813)
                     /codon_start=1
                     /product="human lysosome-associated membrane glycoprotein 
                     1"
                     /label=LAMP1
                     /translation="MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMAN
                     FSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNF
                     TRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHM
                     NNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKY
                     NVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSE
                     GTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNA
                     EEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLIV
                     LIAYLVGRKRSHAGYQTI"
     primer_bind     complement(1980..1998)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2248..2267
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2736..2857
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2773..2792)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2827..2846
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2864..3319)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3001..3022)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3213..3232
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3346..3450
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3452..3809
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3452..3643
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3475..3495)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3660..3795
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3844..4638
                     /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             3844..4638
                     /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(3898..3917)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4508..4527
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4826..4845)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4870..4917
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5246..5834
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5735..5754
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
       61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      541 gatccgctag cgtgccgcca ccatggcggc ccccggcagc gcccggcgac ccctgctgct
      601 gctactgctg ttgctgctgc tcggcctcat gcattgtgcg tcagcagcaa tgtttatggt
      661 gaaaaatggc aacgggaccg cgtgcataat ggccaacttc tctgctgcct tctcagtgaa
      721 ctacgacacc aagagtggcc ctaagaacat gacctttgac ctgccatcag atgccacagt
      781 ggtgctcaac cgcagctcct gtggaaaaga gaacacttct gaccccagtc tcgtgattgc
      841 ttttggaaga ggacatacac tcactctcaa tttcacgaga aatgcaacac gttacagcgt
      901 ccagctcatg agttttgttt ataacttgtc agacacacac cttttcccca atgcgagctc
      961 caaagaaatc aagactgtgg aatctataac tgacatcagg gcagatatag ataaaaaata
     1021 cagatgtgtt agtggcaccc aggtccacat gaacaacgtg accgtaacgc tccatgatgc
     1081 caccatccag gcgtaccttt ccaacagcag cttcagcagg ggagagacac gctgtgaaca
     1141 agacaggcct tccccaacca cagcgccccc tgcgccaccc agcccctcgc cctcacccgt
     1201 gcccaagagc ccctctgtgg acaagtacaa cgtgagcggc accaacggga cctgcctgct
     1261 ggccagcatg gggctgcagc tgaacctcac ctatgagagg aaggacaaca cgacggtgac
     1321 aaggcttctc aacatcaacc ccaacaagac ctcggccagc gggagctgcg gcgcccacct
     1381 ggtgactctg gagctgcaca gcgagggcac caccgtcctg ctcttccagt tcgggatgaa
     1441 tgcaagttct agccggtttt tcctacaagg aatccagttg aatacaattc ttcctgacgc
     1501 cagagacccc gcctttaaag ctgccaacgg ctccctgcga gcgctgcagg ccacagtcgg
     1561 caattcctac aagtgcaacg cggaggagca cgtccgtgtc acgaaggcgt tttcagtcaa
     1621 tatattcaaa gtgtgggtcc aggctttcaa ggtggaaggt ggccagtttg gctctgtgga
     1681 ggagtgtctg ctggacgaga acagcatgct gatccccatc gctgtgggtg gtgccctggc
     1741 ggggctggtc ctcatcgtcc tcatcgccta cctcgtcggc aggaagagga gtcacgcagg
     1801 ctaccagact atcggtggtg gtggtagtgg tggtggtggt agtgctatgg tgagcaagag
     1861 cgaggaggtc atcaaagagt tcatgcgctt caaggtgcgc ttggagggct ccatgaacgg
     1921 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     1981 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca
     2041 gatcatgtac ggctccaagg cgtacgtgaa gcaccccgcc gacgtccccg attacatgaa
     2101 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg cacttcgagg acggcggtct
     2161 ggtgaccgtg actcaggaca cctccctgca ggacggcacg ctgatctaca aggtgaagat
     2221 gcgcggcacc aacttccccc ccgacggccc cgtaatgcag aagaagacct tgggctggga
     2281 ttatgccacc gagcgcctgt accccgaaga cggcgtgctg aagggcgagc ttctggggcg
     2341 cctgaagctg aaggacggcg gcctcaacct ggtggagttc aagaccatct acatggccaa
     2401 gaagcccgtg caactgcccg gctactactt cgtggacacc aagctgggca tcacctccca
     2461 caacgaggac tacaccatcg tggaacagta cgagcgctcc gagggccgcc accacctggg
     2521 catggacgag ctgtacaagg gtacctaata gaagcttggt aaaccgctga tcagcctcga
     2581 ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca
     2641 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2701 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2761 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2821 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     2881 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     2941 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     3001 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     3061 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     3121 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     3181 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     3241 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     3301 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3361 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3421 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     3481 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3541 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3601 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3661 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3721 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3781 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     3841 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3901 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3961 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4021 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4081 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4141 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4201 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4261 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4321 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     4381 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4441 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4501 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4561 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4621 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4681 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4741 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4801 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4861 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     4921 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     4981 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     5041 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     5101 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5161 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5221 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5281 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5341 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5401 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5461 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5521 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5581 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5641 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5701 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5761 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5821 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5881 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5941 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag ccc
//
