LOCUS Exported 5961 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Localization: Lysosome Membrane, Excitation: 515, Emission: 528. ACCESSION . VERSION . KEYWORDS mVenus-Lysosomes-M-20 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5961) TITLE Davidson Yellow fluorescent proteins REFERENCE 2 (bases 1 to 5961) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5961 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 78..381 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 382..585 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 536..556 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 637..646 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 641..650 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 643..1863 /codon_start=1 /product="rat lysosome-associated membrane glycoprotein 1" /label=LAMP1 /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR SHAGYQTI" regulatory 1918..1927 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1924..2643 /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(1969..1990) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2230..2249) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2577..2598 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2766..2887 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2803..2822) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2857..2876 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2894..3349) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3031..3052) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3243..3262 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3376..3480 /gene="bla" /label=AmpR promoter promoter 3482..3839 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3505..3525) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3690..3825 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3752..3771 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3874..4668 /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(3928..3947) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4538..4557 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4856..4875) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4900..4947 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5276..5864 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5765..5784 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc 61 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc 121 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg 181 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat 241 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc 301 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta 361 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag 421 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt 481 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa 541 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt 601 cagatccgct agcgctaccg gactcagatc tcgagcgcca ccatggcggc cccgggcgcc 661 cggcgtccgc tgctcctgtt gctgctggca ggccttgcac acagcgcccc agcactgttc 721 gaggtgaaag acaacaacgg cacagcgtgt ataatggcca gcttctctgc ctcctttctg 781 accacctatg aggctggaca tgtttctaag gtctcgaata tgaccctgcc agcctctgca 841 gaagtcctga agaatagcag ctcttgtggt gaaaagaatg cttctgagcc caccctcgca 901 atcacctttg gagaaggata tttactgaaa ctcaccttca caaaaaacac aacacgttac 961 agtgtccagc acatgtattt cacatataac ctgtcagaca cacaattctt tcccaatgcc 1021 agctccaaag ggcccgacac tgtggattcc acaactgaca tcaaggcaga catcaacaaa 1081 acataccgat gtgtcagcga catcagggtc tacatgaaga atgtgaccat tgtgctctgg 1141 gacgctacta tccaggccta cctgccgagt agcaacttca gcaaggaaga gacacgctgc 1201 ccacaggatc aaccttcccc aactactggg ccacccagcc cctcaccacc acttgtgccc 1261 acaaacccca gtgtgtccaa gtacaatgtg actggtgaca atggaacctg cctgctggcc 1321 tctatggcac tgcaactcaa catcacctac atgaagaagg acaacacgac tgtgaccaga 1381 gcattcaaca tcaacccaag tgacaaatat agtgggactt gcggtgccca gttggtgacc 1441 ctgaaggtgg ggaacaagag cagagtcctg gagctgcagt ttgggatgaa tgccacttct 1501 agcctgtttt tcctgcaagg agttcagttg aacatgactc ttcctgatgc catagagccc 1561 acgttcagca cctccaacta ttccctgaaa gctcttcagg ccagtgtcgg caactcatac 1621 aagtgcaaca gtgaggagca catctttgtc agcaaggcgc tcgccctcaa tgtcttcagc 1681 gtgcaagtcc aggctttcag ggtagaaagt gacaggtttg ggtctgtgga agagtgtgta 1741 caggacggta acaacatgct gatccccatt gctgtgggcg gggccctggc agggctggtc 1801 ctcatcgtcc tcatcgccta cctcatcggc aggaagagga gtcacgcggg ctatcagacc 1861 atctcggaat tcggctccac cggctccacc ggctccaccg gcgcggatcc accggtcgcc 1921 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg 1981 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc 2041 tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc 2101 accctcgtga ccaccctggg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg 2161 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc 2221 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc 2281 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg 2341 cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag 2401 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc 2461 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac 2521 cactacctga gctaccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg 2581 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag 2641 taaagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg 2701 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg 2761 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 2821 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 2881 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt 2941 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa 3001 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag 3061 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt 3121 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac 3181 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag 3241 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg 3301 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact 3361 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg 3421 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt 3481 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag 3541 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg 3601 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 3661 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc 3721 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg 3781 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag 3841 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca 3901 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc 3961 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 4021 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg 4081 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg 4141 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct 4201 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct 4261 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 4321 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa 4381 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc 4441 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt 4501 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct 4561 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 4621 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg 4681 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg 4741 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 4801 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa 4861 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata 4921 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact 4981 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc 5041 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag 5101 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt 5161 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac 5221 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 5281 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 5341 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 5401 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga 5461 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 5521 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 5581 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 5641 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa 5701 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 5761 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 5821 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg 5881 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat 5941 cccctgattc tgtggataac c //