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" primer_bind 227..247 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter complement(272..376) /gene="bla" /label=AmpR promoter rep_origin 403..858 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(490..509) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 700..721 /label=F1ori-F /note="F1 origin, forward primer" polyA_signal 865..986 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(876..895) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 930..949 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS complement(1109..1819) /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 1555..1564 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 1816..1825 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS complement(1841..3091) /codon_start=1 /product="human lysosome-associated membrane glycoprotein 1" /label=LAMP1 /translation="MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMAN FSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNF TRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHM NNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKY NVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSE GTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNA EEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLIV LIAYLVGRKRSHAGYQTI" promoter complement(3168..3371) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(3197..3217) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 3372..3675 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" rep_origin complement(3850..4438) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3930..3949) /label=pBR322ori-F /note="pBR322 origin, forward primer" polyA_signal 4767..4814 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" primer_bind 4839..4858 /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" CDS complement(5046..5840) /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(5157..5176) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 5767..5786 /label=Neo-R /note="Neomycin resistance gene, reverse primer" promoter complement(5875..270) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5889..62 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(5943..5962) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" ORIGIN 1 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg 61 atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata 121 cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc 181 atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac 241 acattccaca gctggttctt tccgcctcag gactcttcct ttttcaatat tattgaagca 301 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 361 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg 421 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc 481 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc 541 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg 601 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg 661 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg 721 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt 781 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa 841 tattaacgct tacaatttac gccttaagat acattgatga gtttggacaa accacaacta 901 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa 961 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg 1021 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg 1081 ctgattatga tctagagtcg cggccgcttt acttgtacag ctcgtccatg cccatcacat 1141 cggtaaaggc cttttgccac tccttgaagt tgagctcggt cttggagtgc ttgagctccg 1201 tcttacggaa cacgtacatc ggctggttct tcagatagtt agccgccatt ggcttggcaa 1261 aggtgtaggt ggtccgcgca gtgctccggt agcgcttgcc atttccagtg gtgtaactcc 1321 acttaaaggt actgatgatg gttttgtcgt tggggtaagt cttcttcgac ctgcaccagt 1381 ccgcagcggt cagcgagttg gtcatcacag gaccgtcagc agggaaacca gtccccttca 1441 cctgggcctc tcctttgatg tggcttccct cgtaggtgta gcggtagtta acagtaaggg 1501 aggcaccatc ttcaaactgc attgtgcgat ggacttggta gccggagcca tctaccatgg 1561 cggcctggaa aggcgacatc ccgtcagggt agggcaggta ctgatggaag ccatacccga 1621 tatgagggac cagaatccag ggggagaact ggaggtcacc cttggtggac ttcaggttta 1681 actcctcata accatcattt ggattgccgg tgccctgacc caccatgtca aagtccacac 1741 cgttgatgga gccaaagatg tgtaactcat gtgtcgctgg gagagaggcc atgttatcct 1801 cctcgccctt gctcaccatg gtggcgaccg gtggatcctt gatagtctgg tagcctgcgt 1861 gactcctctt cctgccgacg aggtaggcga tgaggacgat gaggaccagc cccgccaggg 1921 caccacccac agcgatgggg atcagcatgc tgttctcgtc cagcagacac tcctccacag 1981 agccaaactg gccaccttcc accttgaaag cctggaccca cactttgaat atattgactg 2041 aaaacgcctt cgtgacacgg acgtgctcct ccgcgttgca cttgtaggaa ttgccgactg 2101 tggcctgcag cgctcgcagg gagccgttgg cagctttaaa ggcggggtct ctggcgtcag 2161 gaagaattgt attcaactgg attccttgta ggaaaaaccg gctagaactt gcattcatcc 2221 cgaactggaa gagcaggacg gtggtgccct cgctgtgcag ctccagagtc accaggtggg 2281 cgccgcagct cccgctggcc gaggtcttgt tggggttgat gttgagaagc cttgtcaccg 2341 tcgtgttgtc cttcctctca taggtgaggt tcagctgcag ccccatgctg gccagcaggc 2401 aggtcccgtt ggtgccgctc acgttgtact tgtccacaga ggggctcttg ggcacgggtg 2461 agggcgaggg gctgggtggc gcagggggcg ctgtggttgg ggaaggcctg tcttgttcac 2521 agcgtgtctc tcccctgctg aagctgctgt tggaaaggta cgcctggatg gtggcatcat 2581 ggagcgttac ggtcacgttg ttcatgtgga cctgggtgcc actaacacat ctgtattttt 2641 tatctatatc tgccctgatg tcagttatag attccacagt cttgatttct ttggagctcg 2701 cattggggaa aaggtgtgtg tctgacaagt tataaacaaa actcatgagc tggacgctgt 2761 aacgtgttgc atttctcgtg aaattgagag tgagtgtatg tcctcttcca aaagcaatca 2821 cgagactggg gtcagaagtg ttctcttttc cacaggagct gcggttgagc accactgtgg 2881 catctgatgg caggtcaaag gtcatgttct tagggccact cttggtgtcg tagttcactg 2941 agaaggcagc agagaagttg gccattatgc acgcggtccc gttgccattt ttcaccataa 3001 acattgctgc tgacgcacaa tgcatgaggc cgagcagcag caacagcagt agcagcagca 3061 ggggtcgccg ggcgctgccg ggggccgcca tgtcgactgc agaattcgaa gcttgagctc 3121 gagatctgag tccggtagcg ctagcggatc tgacggttca ctaaaccagc tctgcttata 3181 tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga 3241 cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg 3301 tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa 3361 aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag 3421 tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca 3481 atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt 3541 aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac 3601 gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc 3661 gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat 3721 tactattaat aactaatgca tggcggtaat acggttatcc acagaatcag gggataacgc 3781 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 3841 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 3901 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 3961 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 4021 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 4081 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 4141 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 4201 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 4261 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa 4321 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 4381 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 4441 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 4501 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 4561 aagttttaaa tcaatctaaa gtatatatga gtaacctgag gctatggcag ggcctgccgc 4621 cccgacgttg gctgcgagcc ctgggccttc acccgaactt ggggggtggg gtggggaaaa 4681 ggaagaaacg cgggcgtatt ggccccaatg gggtctcggt ggggtatcga cagagtgcca 4741 gccctgggac cgaaccccgc gtttatgaac aaacgaccca acaccgtgcg ttttattctg 4801 tctttttatt gccgtcatag cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt 4861 tagcctcccc ctagggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat 4921 ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga 4981 atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt 5041 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg 5101 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata 5161 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg 5221 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg 5281 gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa cagttcggct 5341 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc 5401 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga 5461 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca 5521 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc 5581 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat 5641 agccgcgctg cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa 5701 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc 5761 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc 5821 aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc gatctttgca 5881 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc 5941 ggcctcggcc tctgcataaa ta //