LOCUS       sequence-467792-        5971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of lysosome marker LAMP1-Venus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5971)
  AUTHORS   Jang W, Senarath K, Feinberg G, Lu S, Lambert NA
  TITLE     Visualization of endogenous G proteins on endosomes and other 
            organelles.
  JOURNAL   Elife. 2024 Nov 8;13:RP97033. doi: 10.7554/eLife.97033.
  PUBMED    39514269
REFERENCE   2  (bases 1 to 5971)
  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: "Elife."; 
            date: "2024-11-8"; volume: "13:RP97033. doi"; pages: " 
            10.7554/eLife.97033"
COMMENT     Sequence Label: LAMP1-Venus
FEATURES             Location/Qualifiers
     source          1..5971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(1348..1431,1432..2598)
                     /codon_start=1
                     /product="human lysosome-associated membrane glycoprotein 
                     1"
                     /label=LAMP1
                     /translation="MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMAN
                     FSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNF
                     TRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHM
                     NNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKY
                     NVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSE
                     GTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNA
                     EEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLIV
                     LIAYLVGRKRSHAGYQTI"
     regulatory      2614..2623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2620..2622,2623..2625,2626..3339)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2665..2686)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2926..2945)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3273..3294
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3462..3583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3499..3518)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3553..3572
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3590..4045)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3727..3748)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3939..3958
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4072..4176
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4178..4535
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4178..4369
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4201..4221)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4386..4521
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4570..5364
                     /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             4570..5364
                     /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(4624..4643)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5234..5253
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5552..5571)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5596..5643
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact cagatctcga
     1321 gctcaagctt cgaattctgc agtcgacatg gcggcccccg gcagcgcccg gcgacccctg
     1381 ctgctgctac tgctgttgct gctgctcggc ctcatgcatt gtgcgtcagc agcaatgttt
     1441 atggtgaaaa atggcaacgg gaccgcgtgc ataatggcca acttctctgc tgccttctca
     1501 gtgaactacg acaccaagag tggccctaag aacatgacct ttgacctgcc atcagatgcc
     1561 acagtggtgc tcaaccgcag ctcctgtgga aaagagaaca cttctgaccc cagtctcgtg
     1621 attgcttttg gaagaggaca tacactcact ctcaatttca cgagaaatgc aacacgttac
     1681 agcgtccagc tcatgagttt tgtttataac ttgtcagaca cacacctttt ccccaatgcg
     1741 agctccaaag aaatcaagac tgtggaatct ataactgaca tcagggcaga tatagataaa
     1801 aaatacagat gtgttagtgg cacccaggtc cacatgaaca acgtgaccgt aacgctccat
     1861 gatgccacca tccaggcgta cctttccaac agcagcttca gcaggggaga gacacgctgt
     1921 gaacaagaca ggccttcccc aaccacagcg ccccctgcgc cacccagccc ctcgccctca
     1981 cccgtgccca agagcccctc tgtggacaag tacaacgtga gcggcaccaa cgggacctgc
     2041 ctgctggcca gcatggggct gcagctgaac ctcacctatg agaggaagga caacacgacg
     2101 gtgacaaggc ttctcaacat caaccccaac aagacctcgg ccagcgggag ctgcggcgcc
     2161 cacctggtga ctctggagct gcacagcgag ggcaccaccg tcctgctctt ccagttcggg
     2221 atgaatgcaa gttctagccg gtttttccta caaggaatcc agttgaatac aattcttcct
     2281 gacgccagag accccgcctt taaagctgcc aacggctccc tgcgagcgct gcaggccaca
     2341 gtcggcaatt cctacaagtg caacgcggag gagcacgtcc gtgtcacgaa ggcgttttca
     2401 gtcaatatat tcaaagtgtg ggtccaggct ttcaaggtgg aaggtggcca gtttggctct
     2461 gtggaggagt gtctgctgga cgagaacagc atgctgatcc ccatcgctgt gggtggtgcc
     2521 ctggcggggc tggtcctcat cgtcctcatc gcctacctcg tcggcaggaa gaggagtcac
     2581 gcaggctacc agactatcaa ggatccaccg gtcgccacca tggtgagcaa gggcgaggag
     2641 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     2701 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctc
     2761 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctcggctac
     2821 ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     2881 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     2941 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     3001 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     3061 agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     3121 atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc
     3181 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag
     3241 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     3301 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca
     3361 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     3421 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     3481 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3541 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     3601 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3661 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3721 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3781 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3841 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3901 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3961 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4021 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4081 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4141 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     4201 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4261 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4321 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4381 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4441 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     4501 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     4561 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4621 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4681 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4741 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4801 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4861 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4921 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4981 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5041 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     5101 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5161 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5221 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5281 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5341 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5401 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5461 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5521 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     5581 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     5641 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     5701 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     5761 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     5821 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5881 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5941 cagaccccgt agaaaagatc aaaggatctt c
//
