LOCUS       sequence-415450-        5942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dimerization dependent fluorescent protein B 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 5942)
  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 5942)
  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-B
FEATURES             Location/Qualifiers
     source          1..5942
                     /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      562..571
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      566..575
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(568..651,652..1818)
                     /codon_start=1
                     /product="human lysosome-associated membrane glycoprotein 
                     1"
                     /label=LAMP1
                     /translation="MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMAN
                     FSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNF
                     TRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHM
                     NNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKY
                     NVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSE
                     GTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNA
                     EEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLIV
                     LIAYLVGRKRSHAGYQTI"
     primer_bind     complement(1967..1985)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2235..2254
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2429..2452
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2685..2806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2722..2741)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2776..2795
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2813..3268)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2950..2971)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3162..3181
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3295..3399
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3401..3758
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3401..3592
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3424..3444)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3609..3744
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3793..4587
                     /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             3793..4587
                     /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(3847..3866)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4457..4476
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4775..4794)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4819..4866
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5195..5783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5684..5703
                     /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 cgctaccggt cgccaccatg gcggcccccg gcagcgcccg gcgacccctg
      601 ctgctgctac tgctgttgct gctgctcggc ctcatgcatt gtgcgtcagc agcaatgttt
      661 atggtgaaaa atggcaacgg gaccgcgtgc ataatggcca acttctctgc tgccttctca
      721 gtgaactacg acaccaagag tggccctaag aacatgacct ttgacctgcc atcagatgcc
      781 acagtggtgc tcaaccgcag ctcctgtgga aaagagaaca cttctgaccc cagtctcgtg
      841 attgcttttg gaagaggaca tacactcact ctcaatttca cgagaaatgc aacacgttac
      901 agcgtccagc tcatgagttt tgtttataac ttgtcagaca cacacctttt ccccaatgcg
      961 agctccaaag aaatcaagac tgtggaatct ataactgaca tcagggcaga tatagataaa
     1021 aaatacagat gtgttagtgg cacccaggtc cacatgaaca acgtgaccgt aacgctccat
     1081 gatgccacca tccaggcgta cctttccaac agcagcttca gcaggggaga gacacgctgt
     1141 gaacaagaca ggccttcccc aaccacagcg ccccctgcgc cacccagccc ctcgccctca
     1201 cccgtgccca agagcccctc tgtggacaag tacaacgtga gcggcaccaa cgggacctgc
     1261 ctgctggcca gcatggggct gcagctgaac ctcacctatg agaggaagga caacacgacg
     1321 gtgacaaggc ttctcaacat caaccccaac aagacctcgg ccagcgggag ctgcggcgcc
     1381 cacctggtga ctctggagct gcacagcgag ggcaccaccg tcctgctctt ccagttcggg
     1441 atgaatgcaa gttctagccg gtttttccta caaggaatcc agttgaatac aattcttcct
     1501 gacgccagag accccgcctt taaagctgcc aacggctccc tgcgagcgct gcaggccaca
     1561 gtcggcaatt cctacaagtg caacgcggag gagcacgtcc gtgtcacgaa ggcgttttca
     1621 gtcaatatat tcaaagtgtg ggtccaggct ttcaaggtgg aaggtggcca gtttggctct
     1681 gtggaggagt gtctgctgga cgagaacagc atgctgatcc ccatcgctgt gggtggtgcc
     1741 ctggcggggc tggtcctcat cgtcctcatc gcctacctcg tcggcaggaa gaggagtcac
     1801 gcaggctacc agactatcgg tggtggtggt agtggtggtg gtggtagtgc tgccaccatc
     1861 aaagagttca tgcgcttcaa ggtgcgcatg gagggctcca tgaacggcca cgagttcgag
     1921 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
     1981 accaagggcg gccccctgcc ctttgcctgg gacatcctgt ccccccagat catgtacggc
     2041 tccaaagcgt acgtgaggca ccccgccgac atccccgatt acaagaagct gcccttcccc
     2101 gagggcttca agtgggagcg cgtgatgaac ttcgaagacg gcggtctggt gaccgttacc
     2161 caggactcct ccctgcagga cggcacgctg atctgcaagg tgaagattcg cggcaccaac
     2221 ttcccccccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag
     2281 atgctgtacc ccaaggacgg cgtgctgaag ggccatagct atcaggccct gaagctgaag
     2341 gacggcggcc actacctggt ggagttcgag accatctaca tggccaagaa gcccgtgcaa
     2401 ctgcccggcg attactgtgt ggacaccaag ctggacatca cctcccacaa cgaggactac
     2461 accatcgtgg aacagtacgg gcgctccgag ggccgccacc gtctgggcat ggacgagcgg
     2521 tacaagtgac tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga
     2581 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     2641 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     2701 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2761 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     2821 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     2881 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     2941 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3001 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3061 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3121 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3181 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3241 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3301 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3361 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     3421 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3481 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3541 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3601 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3661 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     3721 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     3781 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3841 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3901 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3961 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4021 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4081 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4141 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4201 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4261 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4321 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4381 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4441 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4501 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4561 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4621 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4681 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4741 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     4801 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     4861 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     4921 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     4981 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5041 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5101 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5161 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5221 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5281 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5341 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5401 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5461 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5521 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5581 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5641 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5701 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5761 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5821 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5881 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
     5941 cc
//
