LOCUS Exported 6086 bp ds-DNA circular SYN 07-FEB-2022 DEFINITION Mammalian expression vector for SARS-CoV-2 Nsp15, V5-tagged. Sequence codon-optimized.. ACCESSION . VERSION . KEYWORDS pCG_SARS-CoV2 NSP15 C-V5-tag SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6086) AUTHORS Hayn M, Hirschenberger M, Koepke L, Nchioua R, Straub JH, Klute S, Hunszinger V, Zech F, Prelli Bozzo C, Aftab W, Christensen MH, Conzelmann C, Muller JA, Srinivasachar Badarinarayan S, Sturzel CM, Forne I, Stenger S, Conzelmann KK, Munch J, Schmidt FI, Sauter D, Imhof A, Kirchhoff F, Sparrer KMJ TITLE Systematic functional analysis of SARS-CoV-2 proteins uncovers viral innate immune antagonists and remaining vulnerabilities. JOURNAL Cell Rep. 2021 May 18;35(7):109126. PUBMED 33974846 REFERENCE 2 (bases 1 to 6086) AUTHORS . TITLE Direct Submission JOURNAL Exported Feb 7, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6086 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..304 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 305..508 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 459..479 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 505..529 /label=LNCX /note="Human CMV promoter, forward primer" 5'UTR 598..637 /label=HSV TK 5'-UTR /note="5' untranslated region from the herpes simplex virus thymidine kinase gene" CDS 1714..1755 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" intron 1785..2357 /label=-beta-globin intron /note="intron from rabbit beta-globin gene" primer_bind complement(2489..2508) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 2554..2609 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(2608..2627) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" rep_origin 2963..3098 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(3017..3036) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(3204..3221) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3375..3963) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3455..3474) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4134..4994) /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 4757..4776 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4995..5099) /gene="bla" /label=AmpR promoter primer_bind 5167..5185 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(5223..5245) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 5345..5364 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" rep_origin complement(5431..5886) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5568..5589) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5780..5799 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6012..6034 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 6026..6043 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 6027..6043 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 6050..6069 /label=T7 /note="T7 promoter, forward primer" promoter 6050..6068 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" ORIGIN 1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg 541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccgggggat 601 cttggtggcg tgaaactccc gcacctcttc ggccagcgcc ttgtagaagc gcgtaggcct 661 tctagagcca ccatgtcact tgagaacgtt gcttttaacg tcgtaaataa aggacatttt 721 gatggccaac agggcgaggt tcctgtgtca attattaaca atactgtcta tacaaaggta 781 gacggagtcg atgttgaact ctttgaaaac aagactactc tcccagtgaa tgtggcgttc 841 gaattgtggg cgaagcgtaa cattaaacca gtacccgaag tgaaaatcct caacaatctc 901 ggtgtggata ttgcagccaa taccgtgatc tgggattaca aacgcgacgc tcctgctcac 961 atctctacca taggggtgtg ttccatgact gacatcgcta agaaacccac cgagactata 1021 tgtgcaccac tgacagtatt ctttgacgga agagtagatg gtcaagtgga cctgttccgt 1081 aacgctcgga atggtgtgct gatcactgaa ggtagcgtca agggactcca accgtcagta 1141 ggcccaaagc aagcaagtct gaacggagtg accctgatag gggaagccgt caaaactcaa 1201 tttaattact ataagaaagt tgacggcgtg gtacaacaac ttcctgagac atatttcacc 1261 caaagccgaa atctgcaaga atttaaacca cgttctcaaa tggaaattga cttcctcgaa 1321 ctggccatgg atgagtttat tgagagatac aaattggaag gctatgcctt cgagcatata 1381 gtctacggcg acttttccca ttcacagctt ggcgggctcc atctgcttat agggttggca 1441 aagagattca aagaatcacc tttcgagttg gaagatttca tccccatgga tagcaccgtc 1501 aagaactatt tcattaccga cgcccagacg ggaagcagca agtgcgtctg ctctgtcatc 1561 gatctgctcc tcgatgactt tgtggagata attaaaagcc aagacctcag cgtagttagc 1621 aaagtcgtca aagtgactat agactatact gagatcagct tcatgctctg gtgtaaggat 1681 ggtcacgtcg aaaccttcta ccccaagctg caaggaaaac ctattcccaa cccactgctg 1741 ggtcttgata gcacttagac gcgtcggatc ctgagaactt cagggtgagt ttggggaccc 1801 ttgattgttc tttctttttc gctattgtaa aattcatgtt atatggaggg ggcaaagttt 1861 tcagggtgtt gtttagaatg ggaagatgtc ccttgtatca ccatggaccc tcatgataat 1921 tttgtttctt tcactttcta ctctgttgac aaccattgtc tcctcttatt ttcttttcat 1981 tttctgtaac tttttcgtta aactttagct tgcatttgta acgaattttt aaattcactt 2041 ttgtttattt gtcagattgt aagtactttc tctaatcact tttttttcaa ggcaatcagg 2101 gtatattata ttgtacttca gcacagtttt agagaacaat tgttataatt aaatgataag 2161 gtagaatatt tctgcatata aattctggct ggcgtggaaa tattcttatt ggtagaaaca 2221 actacaccct ggtcatcatc ctgcctttct ctttatggtt acaatgatat acactgtttg 2281 agatgaggat aaaatactct gagtccaaac cgggcccctc tgctaaccat gttcatgcct 2341 tcttctcttt cctacagctc ctgggcaacg tgctggttgt tgtgctgtct catcattttg 2401 gcaaagaatt cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca 2461 atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca 2521 tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa 2581 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt 2641 taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg 2701 ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc 2761 attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg 2821 tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt 2881 tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg 2941 aggagctttt tgcaaaagcc taggcctcca aaaaagcctc ttcactactt ctggaatagc 3001 tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc 3061 ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact 3121 atggttgctg actaattgct gcattaatga atcggccaac gcgcggggag aggcggtttg 3181 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 3241 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 3301 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 3361 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 3421 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 3481 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 3541 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg 3601 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 3661 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 3721 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 3781 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg 3841 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 3901 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 3961 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 4021 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 4081 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 4141 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 4201 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 4261 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 4321 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 4381 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 4441 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 4501 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 4561 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 4621 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 4681 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 4741 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 4801 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 4861 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 4921 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 4981 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 5041 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 5101 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc 5161 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa 5221 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg 5281 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac 5341 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac 5401 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat 5461 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa 5521 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca 5581 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg 5641 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta 5701 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg 5761 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa 5821 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg 5881 gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc 5941 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac 6001 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc 6061 actatagggc gaattcgagc tcgccc //