LOCUS Exported 5085 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Express a EGFP-human MAP1-LC3 in mammalian cells. ACCESSION . VERSION . KEYWORDS pEGFP-hLC3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5085) AUTHORS Tanida I, Tanida-Miyake E, Ueno T, Kominami E TITLE The human homolog of Saccharomyces cerevisiae Apg7p is a Protein-activating enzyme for multiple substrates including human Apg12p, GATE-16, GABARAP, and MAP-LC3. JOURNAL J Biol Chem. 2001 Jan 19;276(3):1701-6. Epub 2000 Nov 28. PUBMED 11096062 REFERENCE 2 (bases 1 to 5085) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5085 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 607..616 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 613..1329 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 613..1329 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(658..679) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(919..938) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1266..1287 /label=EGFP-C /note="EGFP, forward primer" CDS 1345..1719 /codon_start=1 /gene="MAP1LC3B" /product="human LC3 protein" /label=LC3B /note="Atg8 family member found on autophagosomes" /translation="MPSEKTFKQRRTFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLP VLDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESEKD EDGFLYMVYASQETFGMKLSV" polyA_signal 1873..1994 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1910..1929) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1964..1983 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2001..2456) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2138..2159) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2350..2369 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2483..2587 /gene="bla" /label=AmpR promoter promoter 2589..2946 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2612..2632) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2797..2932 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2859..2878 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2981..3775 /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(3035..3054) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3645..3664 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3963..3982) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4007..4054 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4383..4971 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4872..4891 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc 841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac 1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc 1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc 1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 1321 ctgtacaagt ccggactcag atctatgccg tcggagaaga ccttcaagca gcgccgcacc 1381 ttcgaacaaa gagtagaaga tgtccgactt attcgagagc agcatccaac caaaatcccg 1441 gtgataatag aacgatacaa gggtgagaag cagcttcctg ttctggataa aacaaagttc 1501 cttgtacctg accatgtcaa catgagtgag ctcatcaaga taattagaag gcgcttacag 1561 ctcaatgcta atcaggcctt cttcctgttg gtgaacggac acagcatggt cagcgtctcc 1621 acaccaatct cagaggtgta tgagagtgag aaagatgaag atggattcct gtacatggtc 1681 tacgcctccc aggagacgtt cgggatgaaa ttgtcagtgt aagtcgacgg taccgcgggc 1741 ccgggatcca ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt 1801 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca 1861 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 1921 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 1981 atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa 2041 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa 2101 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact 2161 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc 2221 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa 2281 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc 2341 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt 2401 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg 2461 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc 2521 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag 2581 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag 2641 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 2701 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 2761 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 2821 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 2881 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt 2941 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt 3001 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca 3061 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct 3121 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct 3181 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc 3241 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct 3301 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga 3361 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg 3421 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc 3481 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac 3541 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat 3601 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga 3661 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc 3721 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg 3781 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat 3841 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg 3901 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta 3961 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga 4021 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc 4081 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc 4141 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg 4201 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt 4261 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc 4321 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 4381 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 4441 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 4501 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac 4561 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 4621 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 4681 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 4741 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 4801 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 4861 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 4921 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 4981 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct 5041 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat //