LOCUS Exported 9087 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral expression of YFP-tagged LC3. ACCESSION . VERSION . KEYWORDS pLVX-LC3-YFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9087) AUTHORS Flavin WP, Bousset L, Green ZC, Chu Y, Skarpathiotis S, Chaney MJ, Kordower JH, Melki R, Campbell EM TITLE Endocytic vesicle rupture is a conserved mechanism of cellular invasion by amyloid proteins. JOURNAL Acta Neuropathol. 2017 May 19. doi: 10.1007/s00401-017-1722-x. PUBMED 28527044 REFERENCE 2 (bases 1 to 9087) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9087 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 1..634 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 681..806 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1303..1536 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1721..1765 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 2028..2143 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" enhancer 2201..2504 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2505..2708 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2659..2679 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 2747..2756 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2753..3469 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2798..2819) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3059..3078) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3406..3427 /label=EGFP-C /note="EGFP, forward primer" CDS 3485..3859 /codon_start=1 /gene="MAP1LC3B" /product="human LC3 protein" /label=LC3B /note="Atg8 family member found on autophagosomes" /translation="MPSEKTFKQRRTFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLP VLDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESEKD EDGFLYMVYASQETFGMKLSV" promoter 3922..4421 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(3957..3980) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 4354..4373 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 4442..5041 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(4442..4461) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 4938..4958 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 5055..5643 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5108..5128) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5526..5537) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 5850..6483 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" primer_bind complement(6612..6628) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6612..6628) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6625..6647) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6636..6652 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(6660..6690) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6705..6726 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6843..6860) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7014..7602) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7094..7113) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7773..8633) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 8396..8415 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8634..8738) /gene="bla" /label=AmpR promoter polyA_signal 8786..8920 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8823..8842) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8877..8896 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa 2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 2701 gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggtcgcca ccatggtgag 2761 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt 2821 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct 2881 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac 2941 caccttcggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga agcagcacga 3001 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga 3061 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg 3121 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga 3181 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa 3241 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta 3301 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag 3361 ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga 3421 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt ccggactcag 3481 atctatgccg tcggagaaga ccttcaagca gcgccgcacc ttcgaacaaa gagtagaaga 3541 tgtccgactt attcgagagc agcatccaac caaaatcccg gtgataatag aacgatacaa 3601 gggtgagaag cagcttcctg ttctggataa aacaaagttc cttgtacctg accatgtcaa 3661 catgagtgag ctcattaaga taattagaag gcgcttacag ctcaatgcta atcaggcctt 3721 cttcctgttg gtgaacggac acagcatggt cagcgtctcc acaccaatct cagaggtgta 3781 tgagagtgag aaagatgaag atggattcct gtacatggtc tacgcctccc aggagacgtt 3841 cgggatgaaa ttgtcagtgt aagtcgacgg taccgcgggc ccgggatcca ccggatctag 3901 ataactgatc ataattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat 3961 gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac 4021 attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac 4081 cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga 4141 cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc 4201 aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct 4261 gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg 4321 gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt 4381 ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcagc ccaagcttac 4441 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt 4501 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga 4561 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga 4621 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga 4681 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg 4741 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga 4801 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg 4861 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct 4921 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc 4981 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg 5041 accgcgtctg gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc 5101 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg 5161 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc 5221 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg 5281 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg 5341 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga 5401 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct 5461 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg 5521 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc 5581 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc 5641 cgcctggaat taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa 5701 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg 5761 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct 5821 tagccacttt ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca 5881 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta 5941 cacaccaggg ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc 6001 agttgagcca gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc 6061 tgtgagcctg catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag 6121 ccgcctagca tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg 6181 atatcgagct tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg 6241 cgggactggg gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta 6301 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc 6361 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt 6421 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta 6481 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat 6541 cagagagtga gaggccttga cattgctagc gttttaccgt cgacctctag ctagagcttg 6601 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 6661 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 6721 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 6781 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 6841 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 6901 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 6961 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 7021 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 7081 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 7141 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 7201 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 7261 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 7321 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 7381 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 7441 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 7501 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 7561 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 7621 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 7681 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 7741 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 7801 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 7861 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 7921 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 7981 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 8041 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 8101 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 8161 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 8221 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 8281 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 8341 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 8401 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 8461 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 8521 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 8581 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 8641 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 8701 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 8761 cctgacgtcg acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata 8821 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg 8881 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa 8941 ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt 9001 tcatttactc taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg 9061 ttaaatatgt actacaaact tagtagt //