LOCUS Exported 5960 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Localization: Lysosome Membrane, Excitation: 487, Emission: 509. ACCESSION . VERSION . KEYWORDS mEmerald-Lysosomes-20 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5960) TITLE Davidson mEmerald REFERENCE 2 (bases 1 to 5960) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5960 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 104..407 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 408..611 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 562..582 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 663..672 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 667..676 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 669..1889 /codon_start=1 /product="rat lysosome-associated membrane glycoprotein 1" /label=LAMP1 /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR SHAGYQTI" regulatory 1944..1953 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1950..2669 /codon_start=1 /product="brightly fluorescent monomeric variant of GFP" /label=mEmerald /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1995..2016) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2256..2275) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2603..2624 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2791..2912 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2828..2847) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2882..2901 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2919..3374) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3056..3077) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3268..3287 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3401..3505 /gene="bla" /label=AmpR promoter promoter 3507..3864 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3530..3550) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3715..3850 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3777..3796 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3899..4693 /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(3953..3972) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4563..4582 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4881..4900) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4925..4972 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5301..5889 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5790..5809 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat 61 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg 121 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt 181 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac 241 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg 301 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact 361 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt 421 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc 481 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc 541 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata 601 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact cagatctcga 661 gcgccaccat ggcggccccg ggcgcccggc gtccgctgct cctgttgctg ctggcaggcc 721 ttgcacacag cgccccagca ctgttcgagg tgaaagacaa caacggcaca gcgtgtataa 781 tggccagctt ctctgcctcc tttctgacca cctatgaggc tggacatgtt tctaaggtct 841 cgaatatgac cctgccagcc tctgcagaag tcctgaagaa tagcagctct tgtggtgaaa 901 agaatgcttc tgagcccacc ctcgcaatca cctttggaga aggatattta ctgaaactca 961 ccttcacaaa aaacacaaca cgttacagtg tccagcacat gtatttcaca tataacctgt 1021 cagacacaca attctttccc aatgccagct ccaaagggcc cgacactgtg gattccacaa 1081 ctgacatcaa ggcagacatc aacaaaacat accgatgtgt cagcgacatc agggtctaca 1141 tgaagaatgt gaccattgtg ctctgggacg ctactatcca ggcctacctg ccgagtagca 1201 acttcagcaa ggaagagaca cgctgcccac aggatcaacc ttccccaact actgggccac 1261 ccagcccctc accaccactt gtgcccacaa accccagtgt gtccaagtac aatgtgactg 1321 gtgacaatgg aacctgcctg ctggcctcta tggcactgca actcaacatc acctacatga 1381 agaaggacaa cacgactgtg accagagcat tcaacatcaa cccaagtgac aaatatagtg 1441 ggacttgcgg tgcccagttg gtgaccctga aggtggggaa caagagcaga gtcctggagc 1501 tgcagtttgg gatgaatgcc acttctagcc tgtttttcct gcaaggagtt cagttgaaca 1561 tgactcttcc tgatgccata gagcccacgt tcagcacctc caactattcc ctgaaagctc 1621 ttcaggccag tgtcggcaac tcatacaagt gcaacagtga ggagcacatc tttgtcagca 1681 aggcgctcgc cctcaatgtc ttcagcgtgc aagtccaggc tttcagggta gaaagtgaca 1741 ggtttgggtc tgtggaagag tgtgtacagg acggtaacaa catgctgatc cccattgctg 1801 tgggcggggc cctggcaggg ctggtcctca tcgtcctcat cgcctacctc atcggcagga 1861 agaggagtca cgcgggctat cagaccatct cggaattcgg ctccaccggc tccaccggct 1921 ccaccggcgc ggatccaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg 1981 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt 2041 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca 2101 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttgacctac ggcgtgcagt 2161 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg 2221 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg 2281 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact 2341 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaagg 2401 tctatatcac cgccgacaag cagaagaacg gcatcaaggt gaacttcaag acccgccaca 2461 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg 2521 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag ctgagcaaag 2581 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca 2641 ctctcggcat ggacgagctg tacaagtaag cggccgcgac tctagatcat aatcagccat 2701 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg 2761 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac 2821 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 2881 tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt 2941 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat 3001 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt 3061 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt 3121 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag 3181 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg 3241 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc 3301 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc 3361 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt 3421 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct 3481 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt 3541 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 3601 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg 3661 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg 3721 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt 3781 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt 3841 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat 3901 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg 3961 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc 4021 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca 4081 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct 4141 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga 4201 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg 4261 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat 4321 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga 4381 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg 4441 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg 4501 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat 4561 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct 4621 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga 4681 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg 4741 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt 4801 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc 4861 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct 4921 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc 4981 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa 5041 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg 5101 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt 5161 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat 5221 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta 5281 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa 5341 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt 5401 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag 5461 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta 5521 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca 5581 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag 5641 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa 5701 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga 5761 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc 5821 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc 5881 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt 5941 gctcacatgt tctttcctgc //