LOCUS Exported 5962 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION In vivo visualization of lysosomes (can be used for colocalization studies). ACCESSION . VERSION . KEYWORDS Lamp1-mNeonGreen SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5962) AUTHORS Chertkova AO, Mastop M, Postma M, van Bommel N, van der Niet S, Batenburg KL, Joosen L, Gadella TWJ, Okada Y, Goedhart J TITLE Robust and Bright Genetically Encoded Fluorescent Markers for Highlighting Structures and Compartments in Mammalian Cells JOURNAL bioRxiv 160374 REFERENCE 2 (bases 1 to 5962) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5962 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 645..1895 /codon_start=1 /product="human lysosome-associated membrane glycoprotein 1" /label=LAMP1 /translation="MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMAN FSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNF TRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHM NNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKY NVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSE GTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNA EEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLIV LIAYLVGRKRSHAGYQTI" regulatory 1911..1920 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1917..2627 /codon_start=1 /product="bright monomeric yellow-green fluorescent protein derived from LanYFP (Shaner et al., 2013)" /label=mNeonGreen /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ KAFTDVMGMDELYK" regulatory 2172..2181 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" polyA_signal 2750..2871 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2787..2806) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2841..2860 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2878..3333) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3015..3036) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3227..3246 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3360..3464 /gene="bla" /label=AmpR promoter promoter 3466..3823 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3489..3509) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3674..3809 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3736..3755 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3858..4652 /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(3912..3931) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4522..4541 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4840..4859) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4884..4931 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5260..5848 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5749..5768 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggactcag atctcgagct caagcttcga attctgcagt cgacatggcg gcccccggca 661 gcgcccggcg acccctgctg ctgctactgc tgttgctgct gctcggcctc atgcattgtg 721 cgtcagcagc aatgtttatg gtgaaaaatg gcaacgggac cgcgtgcata atggccaact 781 tctctgctgc cttctcagtg aactacgaca ccaagagtgg ccctaagaac atgacctttg 841 acctgccatc agatgccaca gtggtgctca accgcagctc ctgtggaaaa gagaacactt 901 ctgaccccag tctcgtgatt gcttttggaa gaggacatac actcactctc aatttcacga 961 gaaatgcaac acgttacagc gtccagctca tgagttttgt ttataacttg tcagacacac 1021 accttttccc caatgcgagc tccaaagaaa tcaagactgt ggaatctata actgacatca 1081 gggcagatat agataaaaaa tacagatgtg ttagtggcac ccaggtccac atgaacaacg 1141 tgaccgtaac gctccatgat gccaccatcc aggcgtacct ttccaacagc agcttcagca 1201 ggggagagac acgctgtgaa caagacaggc cttccccaac cacagcgccc cctgcgccac 1261 ccagcccctc gccctcaccc gtgcccaaga gcccctctgt ggacaagtac aacgtgagcg 1321 gcaccaacgg gacctgcctg ctggccagca tggggctgca gctgaacctc acctatgaga 1381 ggaaggacaa cacgacggtg acaaggcttc tcaacatcaa ccccaacaag acctcggcca 1441 gcgggagctg cggcgcccac ctggtgactc tggagctgca cagcgagggc accaccgtcc 1501 tgctcttcca gttcgggatg aatgcaagtt ctagccggtt tttcctacaa ggaatccagt 1561 tgaatacaat tcttcctgac gccagagacc ctgcctttaa agctgccaac ggctccctgc 1621 gagcgctgca ggccacagtc ggcaattcct acaagtgcaa cgcggaggag cacgtccgtg 1681 tcacgaaggc gttttcagtc aatatattca aagtgtgggt ccaggctttc aaggtggaag 1741 gtggccagtt tggctctgtg gaggagtgtc tgctggacga gaacagcatg ctgatcccca 1801 tcgctgtggg tggtgccctg gcggggctgg tcctcatcgt cctcatcgcc tacctcgtcg 1861 gcaggaagag gagtcacgca ggctaccaga ctatcaagga tccaccggtc gccaccatgg 1921 tgagcaaggg cgaggaggat aacatggcct ctctcccagc gacacatgag ttacacatct 1981 ttggctccat caacggtgtg gactttgaca tggtgggtca gggcaccggc aatccaaatg 2041 atggttatga ggagttaaac ctgaagtcca ccaagggtga cctccagttc tccccctgga 2101 ttctggtccc tcatatcggg tatggcttcc atcagtacct gccctaccct gacgggatgt 2161 cgcctttcca ggccgccatg gtagatggct ccggctacca agtccatcgc acaatgcagt 2221 ttgaagatgg tgcctccctt actgttaact accgctacac ctacgaggga agccacatca 2281 aaggagaggc ccaggtgaag gggactggtt tccctgctga cggtcctgtg atgaccaact 2341 cgctgaccgc tgcggactgg tgcaggtcga agaagactta ccccaacgac aaaaccatca 2401 tcagtacctt taagtggagt tacaccactg gaaatggcaa gcgctaccgg agcactgcgc 2461 ggaccaccta cacctttgcc aagccaatgg cggctaacta tctgaagaac cagccgatgt 2521 acgtgttccg taagacggag ctcaagcact ccaagaccga gctcaacttc aaggagtggc 2581 aaaaggcctt taccgatgtg atgggcatgg acgagctgta caagtaaagc ggccgcgact 2641 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc 2701 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat 2761 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 2821 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa 2881 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt 2941 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag 3001 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg 3061 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat 3121 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc 3181 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga 3241 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac 3301 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc 3361 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac 3421 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa 3481 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag 3541 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc 3601 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc 3661 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg 3721 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca 3781 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag 3841 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt 3901 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg 3961 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg 4021 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg 4081 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg 4141 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca 4201 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc 4261 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc 4321 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca 4381 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga 4441 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg 4501 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg 4561 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg 4621 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga 4681 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag 4741 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct 4801 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat 4861 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg 4921 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc 4981 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag 5041 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc 5101 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 5161 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 5221 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 5281 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 5341 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 5401 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 5461 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 5521 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 5581 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 5641 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 5701 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 5761 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 5821 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 5881 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 5941 taaccgtatt accgccatgc at //