LOCUS Exported 6188 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION LAMP1 (Lysosome Membrane) labeling with near-infrared fluorescent protein miRFP703. ACCESSION . VERSION . KEYWORDS pLAMP1-miRFP703 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6188) AUTHORS Shcherbakova DM, Baloban M, Emelyanov AV, Brenowitz M, Guo P, Verkhusha VV TITLE Bright monomeric near-infrared fluorescent proteins as tags and biosensors for multiscale imaging. JOURNAL Nat Commun. 2016 Aug 19;7:12405. doi: 10.1038/ncomms12405. PUBMED 27539380 REFERENCE 2 (bases 1 to 6188) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6188 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 68..371 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 372..575 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 526..546 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 627..636 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 631..640 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 633..1853 /codon_start=1 /product="rat lysosome-associated membrane glycoprotein 1" /label=LAMP1 /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR SHAGYQTI" CDS 1914..2861 /codon_start=1 /product="bright monomeric near-infrared fluorescent protein engineered from a bacterial phytochrome, with an emission maximum of 703 nm (Shcherbakova et al., 2016)" /label=miRFP703 /label=ORF frame 1 /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSLVPQMARQLYVRQRVRVLVDVT YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPIHLQFLKDMGVRATLAVSLVVGGKLWGLV VCHHYLPRFIRFELRAICKRLAERIATRITALES" polyA_signal 2983..3104 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3020..3039) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3074..3093 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3111..3566) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3248..3269) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3460..3479 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3593..3697 /gene="bla" /label=AmpR promoter promoter 3699..4056 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3722..3742) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3907..4042 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3969..3988 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4091..4885 /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(4145..4164) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4755..4774 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5073..5092) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5117..5164 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5493..6081 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5982..6001 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg 61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc 121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt 181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc 241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg 301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg 361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact 421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa 481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta 541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct 601 agcgctaccg gactcagatc tcgagcgcca ccatggcggc cccgggcgcc cggcggccgc 661 tgctcctgtt gctgctggca ggccttgcac acagcgcccc agcactgttc gaggtgaaag 721 acaacaacgg cacagcgtgt ataatggcca gcttctctgc ctcctttctg accacctatg 781 aggctggaca tgtttctaag gtctcgaata tgaccctgcc agcctctgca gaagtcctga 841 agaatagcag ctcttgtggt gaaaagaatg cttctgagcc caccctcgca atcacctttg 901 gagaaggata tttactgaaa ctcaccttca caaaaaacac aacacgttac agtgtccagc 961 acatgtattt cacatataac ctgtcagaca cacaattctt tcccaatgcc agctccaaag 1021 ggcccgacac tgtggattcc acaactgaca tcaaggcaga catcaacaaa acataccgat 1081 gtgtcagcga catcagggtc tacatgaaga atgtgaccat tgtgctctgg gacgctacta 1141 tccaggccta cctgccgagt agcaacttca gcaaggaaga gacacgctgc ccacaggatc 1201 aaccttcccc aactactggg ccacccagcc cctcaccacc acttgtgccc acaaacccca 1261 gtgtgtccaa gtacaatgtg actggtgaca atggaacctg cctgctggcc tctatggcac 1321 tgcaactcaa catcacctac atgaagaagg acaacacgac tgtgaccaga gcattcaaca 1381 tcaacccaag tgacaaatat agtgggactt gcggtgccca gttggtgacc ctgaaggtgg 1441 ggaacaagag cagagtcctg gagctgcagt ttgggatgaa tgccacttct agcctgtttt 1501 tcctgcaagg agttcagttg aacatgactc ttcctgatgc catagagccc acgttcagca 1561 cctccaacta ttccctgaaa gctcttcagg ccagtgtcgg caactcatac aagtgcaaca 1621 gtgaggagca catctttgtc agcaaggcgc tcgccctcaa tgtcttcagc gtgcaagtcc 1681 aggctttcag ggtagaaagt gacaggtttg ggtctgtgga agagtgtgta caggacggta 1741 acaacatgct gatccccatt gctgtgggcg gggccctggc agggctggtc ctcatcgtcc 1801 tcatcgccta cctcatcggc aggaagagga gtcacgcggg ctatcagacc atctcggaat 1861 tcggctccac cggctccacc ggctccaccg gcgcggatcc accggtggct accatggtag 1921 caggtcatgc ctctggcagc cccgcattcg ggaccgcctc tcattcgaat tgcgaacatg 1981 aagagatcca cctcgccggc tcgatccagc cgcatggcgc gcttctggtc gtcagcgaac 2041 atgatcatcg cgtcatccag gccagcgcca acgccgcgga atttctgaat ctcggaagcg 2101 tactcggcgt tccgctcgcc gagatcgacg gcgatctgtt gatcaagatc ctgccgcatc 2161 tcgatcccac cgccgaaggc atgccggtcg cggtgcgctg ccggatcggc aatccctcta 2221 cggagtactg cggtctgatg catcggcctc cggaaggcgg gctgatcatc gaactcgaac 2281 gtgccggccc gtcgatcgat ctgtcaggca cgctggcgcc ggcgctggag cggatccgca 2341 cggcgggttc actgcgcgcg ctgtgcgatg acaccgtgct gctgtttcag cagtgcaccg 2401 gctacgaccg ggtgatggtg tatcgtttcg atgagcaagg ccacggcctg gtattctccg 2461 agtgccatgt gcctgggctc gaatcctatt tcggcaaccg ctatccgtcg tcgctggtcc 2521 cgcagatggc gcggcagctg tacgtgcggc agcgcgtccg cgtgctggtc gacgtcacct 2581 atcagccggt gccgctggag ccgcggctgt cgccgctgac cgggcgcgat ctcgacatgt 2641 cgggctgctt cctgcgctcg atgtcgccga tccatctgca gttcctgaag gacatgggcg 2701 tgcgcgccac cctggcggtg tcgctggtgg tcggcggcaa gctgtggggc ctggttgtct 2761 gtcaccatta tctgccgcgc ttcatccgtt tcgagctgcg ggcgatctgc aaacggctcg 2821 ccgaaaggat cgcgacgcgg atcaccgcgc ttgagagcta agcggccgcg actctagatc 2881 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc 2941 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct 3001 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 3061 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag 3121 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca 3181 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag 3241 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg 3301 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt 3361 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag 3421 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc 3481 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc 3541 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac 3601 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc 3661 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg 3721 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg 3781 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca 3841 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact 3901 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 3961 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag 4021 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg 4081 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga 4141 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt 4201 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct 4261 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg 4321 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt 4381 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc 4441 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc 4501 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga 4561 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag 4621 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat 4681 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg 4741 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc 4801 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta 4861 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg 4921 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc 4981 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg 5041 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa 5101 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt 5161 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc 5221 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt 5281 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac 5341 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag 5401 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg 5461 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc 5521 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag 5581 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt 5641 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac 5701 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc 5761 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt 5821 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt 5881 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc 5941 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt 6001 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca 6061 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt 6121 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt 6181 attaccgc //