LOCUS Exported 6835 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of human UBXD7 with C-terminal strep-HA tag. ACCESSION . VERSION . KEYWORDS pcDNA5FRT/TO-UBXD7-Strep-HA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6835) AUTHORS Hulsmann J, Kravic B, Weith M, Gstaiger M, Aebersold RH, Collins BC, Meyer H TITLE AP-SWATH reveals direct involvement of VCP/p97 in integrated stress response signaling through facilitating CReP/PPP1R15B degradation. JOURNAL Mol Cell Proteomics. 2018 Mar 29. pii: RA117.000471. doi: 10.1074/mcp.RA117.000471. PUBMED 29599191 REFERENCE 2 (bases 1 to 6835) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6835 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 820..838 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 841..859 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 869..893 /label=LNCX /note="Human CMV promoter, forward primer" protein_bind 1036..1060 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2150..2161 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" protein_bind complement(2534..2558) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2605..2628 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 2665..2688 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 2719..2745 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(2788..2805) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2794..3018 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(3151..3170) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 3302..3349 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS 3357..4379 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " polyA_signal 4509..4630 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4546..4565) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4600..4619 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4679..4695) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4679..4695) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4692..4714) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4703..4719 /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(4727..4757) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4772..4793 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4910..4927) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5081..5669) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5161..5180) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6138..6515) /label=ISS /note="immunostimulatory sequence from the AmpR gene; contains unmethylated CpG dinucleotides in the context of 5'-AACGTT-3' (Sato et al., 1996)" primer_bind 6462..6481 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6700..6804) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc 841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga 901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga 961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga 1021 attctgcaga tatcaacaag tttgtacaaa aaagcaggct ccaccatggc tgcccacggg 1081 ggctccgcgg cgtcctcggc gctgaagggg ttaattcaac agttcaccac cattaccggt 1141 gcaagtgaaa gtgtaggaaa acatatgctt gaagcgtgca acaataatct ggaaatggca 1201 gtcactatgt ttttggatgg tggaggaatc gctgaagagc ccagtaccag ttcagcaagt 1261 gtctctactg tcagaccaca cacagaagaa gaagttcgtg ccccaattcc tcaaaagcag 1321 gaaatactgg tggaaccaga accattattt ggtgctccta aaagacgacg gcctgcacgt 1381 tcaatttttg atggtttccg ggattttcag actgaaacta ttcggcaaga acaagaatta 1441 agaaatggag gagctatcga taagaaatta actacccttg cagatctatt ccggccaccc 1501 attgatttga tgcataaagg cagctttgaa acagccaaag agtgtggcca gatgcaaaat 1561 aagtggctga tgataaacat tcaaaatgtt caagactttg catgtcagtg cctcaaccgc 1621 gatgtgtgga gcaacgaagc tgtgaagaat attatccggg aacatttcat tttctggcag 1681 gtttatcatg acagtgagga aggtcagaga tacatacagt tttataagtt aggggatttc 1741 ccctatgttt ccatattgga cccacggaca ggtcagaagc tagtagaatg gcaccagtta 1801 gatgtatctt ctttcttgga ccaagtgacg ggatttctgg gtgaacatgg acaactggat 1861 ggactttcta gcagtccccc caaaaaatgt gcccgttccg agagccttat agatgcaagt 1921 gaagacagcc agctagaagc tgccatcaga gcctccttac aagaaacaca ttttgattca 1981 acacagacaa aacaggatag ccgctcagat gaagaatctg aatctgaact tttttctggc 2041 agtgaggagt tcatatccgt ttgtggctct gatgaagaag aagaggtaga gaatcttgcc 2101 aagtccagaa agtctcccca caaagatttg gggcatagaa aagaggagaa tagaaggccg 2161 ctgactgagc caccagtcag aactgatcct ggaacagcca caaaccacca aggattgcca 2221 gctgtggatt cagagatact ggagatgcca cctgaaaaag cagatggagt agtggagggg 2281 atagatgtaa atggaccaaa agcacagctg atgttgcggt atccagatgg aaaaagggaa 2341 cagatcactc ttccagagca agctaaactg ctagctttgg tgaagcacgt gcagtctaaa 2401 ggatacccaa atgaacgttt tgaacttctc accaactttc ctcgaaggaa attatctcat 2461 ctggactatg atattacatt gcaagaggca ggcctttgtc ctcaagagac tgtctttgta 2521 caggaaagaa attacccagc tttcttgtac aaagtggttg atatcaaggc cgccgacatc 2581 accagcctgt acaagaaagt ggggtggagc catccccagt tcgagaaggg cggaggatct 2641 ggcggcggaa gcggcggagg aagctggtcc caccctcagt ttgagaaagg caccgagctg 2701 ggcagcacca tggccagcta cccctacgac gtgcccgact acgcctgatg actcgagtct 2761 agagggcccg ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg 2821 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt 2881 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg 2941 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg 3001 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc 3061 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 3121 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg 3181 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat 3241 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtacct 3301 agaagttcct attccgaagt tcctattctc tagaaagtat aggaacttcc ttggccaaaa 3361 agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct 3421 ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag 3481 ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg 3541 tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggaat 3601 tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc 3661 tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg gatgcgatcg 3721 ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc 3781 aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc 3841 aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc 3901 tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca 3961 atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg 4021 gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg 4081 agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc 4141 gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt 4201 tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga 4261 ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag 4321 aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaatagc 4381 acgtactacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg 4441 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg 4501 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 4561 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 4621 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt 4681 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 4741 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 4801 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 4861 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 4921 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 4981 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 5041 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 5101 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 5161 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 5221 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 5281 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 5341 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 5401 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 5461 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 5521 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 5581 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 5641 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 5701 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 5761 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 5821 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 5881 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 5941 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 6001 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 6061 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 6121 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 6181 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 6241 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 6301 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 6361 catccgtaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 6421 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 6481 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 6541 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 6601 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 6661 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 6721 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 6781 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc //