LOCUS Exported 9353 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral expression of human TDP-43 A90V disease mutant in mammalian cells.. ACCESSION . VERSION . KEYWORDS pLVX-Puro-TDP-43-A90V SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9353) AUTHORS Roczniak-Ferguson A, Ferguson SM TITLE Pleiotropic requirements for human TDP-43 in the regulation of cell and organelle homeostasis. JOURNAL Life Sci Alliance. 2019 Sep 16;2(5). pii: 2/5/e201900358. doi: 10.26508/lsa.201900358. Print 2019 Oct. PUBMED 31527135 REFERENCE 2 (bases 1 to 9353) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9353 /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" primer_bind 2705..2729 /label=LNCX /note="Human CMV promoter, forward primer" promoter 4141..4640 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(4176..4199) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 4573..4592 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 4661..5260 /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(4661..4680) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 5157..5177 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 5274..5862 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5327..5347) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5745..5756) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 6069..6702 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" primer_bind complement(6831..6847) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6831..6847) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6844..6866) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6855..6871 /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(6879..6909) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6924..6945 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7062..7079) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7233..7821) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7313..7332) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7992..8852) /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 8615..8634 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8853..8957) /gene="bla" /label=AmpR promoter polyA_signal 9005..9139 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(9042..9061) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 9096..9115 /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 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag 2821 ctcaagcttc gaattctgca gtcgacggta ccgcgggccc gccaccatgt ctgaatatat 2881 tcgggtaacc gaagatgaga acgatgagcc cattgaaata ccatcggaag acgatgggac 2941 ggtgctgctc tccacggtta cagcccagtt tccaggggcg tgtgggcttc gctacaggaa 3001 tccagtgtct cagtgtatga gaggtgtccg gctggtagaa ggaattctgc atgccccaga 3061 tgctggctgg ggaaatctgg tgtatgttgt caactatcca aaagataaca aaagaaaaat 3121 ggatgagaca gatgtttcat cagcagtgaa agtgaaaaga gcagtccaga aaacatccga 3181 tttaatagtg ttgggtctcc catggaaaac aaccgaacag gacctgaaag agtattttag 3241 tacctttgga gaagttctta tggtgcaggt caagaaagat cttaagactg gtcattcaaa 3301 ggggtttggc tttgttcgtt ttacggaata tgaaacacaa gtgaaagtaa tgtcacagcg 3361 acatatgata gatggacgat ggtgtgactg caaacttcct aattctaagc aaagccaaga 3421 tgagcctttg agaagcagaa aagtgtttgt ggggcgctgt acagaggaca tgactgagga 3481 tgagctgcgg gagttcttct ctcagtacgg ggatgtgatg gatgtcttca tccccaagcc 3541 attcagggcc tttgcctttg ttacatttgc agatgatcag attgcgcagt ctctttgtgg 3601 agaggacttg atcattaaag gaatcagcgt tcatatatcc aatgccgaac ctaagcacaa 3661 tagcaataga cagttagaaa gaagtggaag atttggtggt aatccaggtg gctttgggaa 3721 tcagggtgga tttggtaata gcagaggggg tggagctggt ttgggaaaca atcaaggtag 3781 taatatgggt ggtgggatga actttggtgc gttcagcatt aatccagcca tgatggctgc 3841 cgcccaggca gcactacaga gcagttgggg tatgatgggc atgttagcca gccagcagaa 3901 ccagtcaggc ccatcgggta ataaccaaaa ccaaggcaac atgcagaggg agccaaacca 3961 ggccttcggt tctggaaata actcttatag tggctctaat tctggtgcag caattggttg 4021 gggatcagca tccaatgcag ggtcgggcag tggttttaat ggaggctttg gctcaagcat 4081 ggattctaag tcttctggct ggggaatgta ggggatcccg cgactctaga taattctacc 4141 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg 4201 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc 4261 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca 4321 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac 4381 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt 4441 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg 4501 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct 4561 ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt 4621 cctcatctcc gggcctttcg acctgcagcc caagcttacc atgaccgagt acaagcccac 4681 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt 4741 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac 4801 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc 4861 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt 4921 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca 4981 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac 5041 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg 5101 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa 5161 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg 5221 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga ccgcgtctgg aacaatcaac 5281 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta 5341 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt 5401 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg 5461 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg 5521 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca 5581 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca 5641 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg 5701 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag 5761 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc 5821 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt aattctgcag 5881 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga 5941 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt 6001 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa 6061 gaggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat 6121 ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc caggggtcag 6181 atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga 6241 agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga 6301 tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt 6361 ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg 6421 actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc 6481 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc 6541 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa 6601 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga 6661 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc 6721 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggccttgac 6781 attgctagcg ttttaccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg 6841 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 6901 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 6961 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 7021 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 7081 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 7141 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 7201 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 7261 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 7321 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 7381 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 7441 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 7501 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 7561 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 7621 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 7681 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 7741 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 7801 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 7861 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 7921 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 7981 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 8041 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 8101 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 8161 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 8221 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 8281 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 8341 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 8401 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 8461 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 8521 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 8581 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 8641 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 8701 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 8761 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 8821 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 8881 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 8941 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga 9001 gatcaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 9061 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 9121 atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc 9181 caccccatac cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga 9241 ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt 9301 agtagttttt aaagaaattg tatttgttaa atatgtacta caaacttagt agt //