LOCUS       sequence-415474-        5987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dimerization dependent fluorescent protein GA 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 5987)
  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 5987)
  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-GA
FEATURES             Location/Qualifiers
     source          1..5987
                     /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"
     primer_bind     2442..2465
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2730..2851
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2767..2786)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2821..2840
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2858..3313)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2995..3016)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3207..3226
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3340..3444
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3446..3803
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3446..3637
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3469..3489)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3654..3789
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3838..4632
                     /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             3838..4632
                     /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(3892..3911)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4502..4521
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4820..4839)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4864..4911
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5240..5828
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5729..5748
                     /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 gggccgccct tacgagggca cccagaccgc
     1981 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccct
     2041 gatcatgtac ggctccaaga tgtacgtgaa gcaccccgcc gacgtccccg attacatgaa
     2101 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg cacttcgagg acggcggtct
     2161 ggtgaccgcg acacaggact cctccctgca ggacggcacg ctgatctaca aggtgaagat
     2221 gcgcggcacc aacttccccc ccgacggccc cgtaatgcag aagaagacct tgggctggga
     2281 ttatgccacc gagcgcctgt accccgaaga aggcgtgctg aagggcgagc tcctggggcg
     2341 cctgaagctg aaggacggcg gcctcaacct ggtggagtcc aagaccatct acatggccaa
     2401 gaagcccgtg caactgcccg gctactactt cgtggacacc aagctggaca tcacctccca
     2461 caacgaggac tacaccatcg tggaacagta cgagcgctcc gagggccgcc accacctggg
     2521 gcatggcacc ggcagcacct aatagaagct tggtaaaccg ctgatcagcc tcgactgcag
     2581 tcgacggtac cgcgggcccg ggatccaccg gatctagata actgatcata atcagccata
     2641 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     2701 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     2761 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2821 gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg
     2881 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     2941 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     3001 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     3061 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     3121 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     3181 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     3241 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     3301 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3361 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3421 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     3481 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3541 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3601 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3661 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3721 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3781 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     3841 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3901 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3961 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     4021 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4081 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4141 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4201 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4261 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4321 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     4381 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4441 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4501 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4561 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4621 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4681 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4741 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4801 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     4861 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     4921 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     4981 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     5041 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     5101 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5161 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5221 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5281 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5341 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5401 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5461 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5521 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5581 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5641 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5701 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5761 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5821 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5881 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     5941 attagttatt aatagtaatc aattacgggg tcattagttc atagccc
//
