LOCUS Exported 5961 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Visualization of lysosome in mammalian cells using mGold. ACCESSION . VERSION . KEYWORDS pCMV-mGold-Lysosome-N-20 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5961) AUTHORS Lee J, Liu Z, Suzuki PH, Ahrens JF, Lai S, Lu X, Guan S, St-Pierre F TITLE Versatile phenotype-activated cell sorting. JOURNAL Sci Adv. 2020 Oct 23;6(43). pii: 6/43/eabb7438. doi: 10.1126/sciadv.abb7438. Print 2020 Oct. PUBMED 33097540 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 1..304 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 305..508 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 459..479 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 560..569 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 564..573 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 566..1786 /codon_start=1 /product="rat lysosome-associated membrane glycoprotein 1" /label=LAMP1 /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR SHAGYQTI" regulatory 1841..1850 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1847..2566 /codon_start=1 /product="photostable derivative of mVenus (Lee et al., 2020)" /label=mGold /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTSLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1892..1913) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2153..2172) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2500..2521 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2689..2810 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2726..2745) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2780..2799 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2817..3272) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2954..2975) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3166..3185 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3299..3403 /gene="bla" /label=AmpR promoter promoter 3405..3762 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3428..3448) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3613..3748 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3675..3694 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3797..4591 /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(3851..3870) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4461..4480 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4779..4798) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4823..4870 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5199..5787 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5688..5707 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 541 ccggactcag atctcgagcg ccaccatggc ggccccgggc gcccggcgtc cgctgctcct 601 gttgctgctg gcaggccttg cacacagcgc cccagcactg ttcgaggtga aagacaacaa 661 cggcacagcg tgtataatgg ccagcttctc tgcctccttt ctgaccacct atgaggctgg 721 acatgtttct aaggtctcga atatgaccct gccagcctct gcagaagtcc tgaagaatag 781 cagctcttgt ggtgaaaaga atgcttctga gcccaccctc gcaatcacct ttggagaagg 841 atatttactg aaactcacct tcacaaaaaa cacaacacgt tacagtgtcc agcacatgta 901 tttcacatat aacctgtcag acacacaatt ctttcccaat gccagctcca aagggcccga 961 cactgtggat tccacaactg acatcaaggc agacatcaac aaaacatacc gatgtgtcag 1021 cgacatcagg gtctacatga agaatgtgac cattgtgctc tgggacgcta ctatccaggc 1081 ctacctgccg agtagcaact tcagcaagga agagacacgc tgcccacagg atcaaccttc 1141 cccaactact gggccaccca gcccctcacc accacttgtg cccacaaacc ccagtgtgtc 1201 caagtacaat gtgactggtg acaatggaac ctgcctgctg gcctctatgg cactgcaact 1261 caacatcacc tacatgaaga aggacaacac gactgtgacc agagcattca acatcaaccc 1321 aagtgacaaa tatagtggga cttgcggtgc ccagttggtg accctgaagg tggggaacaa 1381 gagcagagtc ctggagctgc agtttgggat gaatgccact tctagcctgt ttttcctgca 1441 aggagttcag ttgaacatga ctcttcctga tgccatagag cccacgttca gcacctccaa 1501 ctattccctg aaagctcttc aggccagtgt cggcaactca tacaagtgca acagtgagga 1561 gcacatcttt gtcagcaagg cgctcgccct caatgtcttc agcgtgcaag tccaggcttt 1621 cagggtagaa agtgacaggt ttgggtctgt ggaagagtgt gtacaggacg gtaacaacat 1681 gctgatcccc attgctgtgg gcggggccct ggcagggctg gtcctcatcg tcctcatcgc 1741 ctacctcatc ggcaggaaga ggagtcacgc gggctatcag accatctcgg aattcggctc 1801 caccggctcc accggctcca ccggcgcgga tccaccggtc gccaccatgg tgagcaaggg 1861 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg 1921 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct 1981 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccagcct 2041 gggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt 2101 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg 2161 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga 2221 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa 2281 ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa 2341 cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca 2401 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca 2461 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt 2521 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc 2581 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca 2641 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt 2701 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 2761 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat 2821 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt 2881 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg 2941 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt 3001 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc 3061 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg 3121 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa 3181 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc 3241 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg 3301 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca 3361 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac 3421 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga 3481 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc 3541 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc 3601 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 3661 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 3721 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg 3781 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg 3841 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc 3901 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg 3961 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt 4021 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg 4081 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat 4141 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca 4201 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca 4261 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa 4321 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa 4381 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc 4441 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga 4501 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc 4561 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac 4621 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg 4681 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc 4741 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata 4801 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg 4861 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg 4921 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt 4981 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca 5041 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag 5101 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac 5161 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc 5221 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat 5281 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat 5341 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct 5401 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt 5461 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg 5521 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta 5581 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg 5641 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg 5701 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc 5761 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg 5821 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat 5881 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca 5941 tagcccatat atggagttcc g //