LOCUS Exported 6786 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 ORF6 protein. ACCESSION . VERSION . KEYWORDS pGBW-m4252882 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6786) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6786) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6786 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 188..208 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 209..250 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" polyA_signal 319..440 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(330..349) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 384..403 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 555..601 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 631..767 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 812..883 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1109..1697) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1189..1208) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 2197..3406 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 3243..3262 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" CDS 3410..4438 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA KE" polyA_signal 4465..4513 /note="synthetic polyadenylation signal" CDS 4779..5639 /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 complement(4997..5016) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 5652..5788 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 5940..6011 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 6016..6062 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 6080..6128 /note="synthetic polyadenylation signal" enhancer 6209..6512 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 6514..6716 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 6667..6687 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgttt catctcgttg actttcaggt tactatagca gagatattac taattattat 61 gaggactttt aaagtttcca tttggaatct tgattacatc ataaacctca taattaaaaa 121 tttatctaag tcactaactg agaataaata ttctcaatta gatgaagagc aaccaatgga 181 gattgatgaa aacctttact ttcagtcggg gaaacctata cccaatccac ttctcggctt 241 agattctacc taaggatccg caggcctctg ctagcttgac tgactgagat acagcgtacc 301 ttcagctcac agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc 361 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta 421 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg 481 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat 541 ctctatcggt atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt 601 gtgcccctcg ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg 661 acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa 721 cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat 781 cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc 841 actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt 901 gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg 961 gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc 1021 gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga 1081 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 1141 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 1201 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 1261 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt 1321 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 1381 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 1441 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 1501 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg 1561 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 1621 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 1681 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 1741 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 1801 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 1861 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 1921 catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa 1981 cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc 2041 cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg 2101 aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat 2161 ccgagatccg aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc 2221 ctcccgcggg cgcccccctc gtcacggcga gcgctgccac gtcagacgaa gggcgcagga 2281 gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc 2341 cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc 2401 actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc 2461 tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt 2521 ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg 2581 tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg 2641 ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga 2701 gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag 2761 tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca 2821 tggtgggcgg caagaaccca aggtcttgag cccttcgcta atgcgggaaa gctcttattc 2881 gggtgagatg ggctgggcac catctgggga ccctgacgtg aagtttgtca ctgactggag 2941 aactcggttt gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac 3001 ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta 3061 taatgcaggg tggggccacc agccggtagg tgtgcggtag gcttttctcc gtcgcaggac 3121 gcagggttcg ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg 3181 ggagggataa gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta 3241 gctgaagctc cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt 3301 ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagacttg 3361 taaattgtcc gctaaattct ggccgttttt ggcttttttg ttagacaaca tgggtaaaaa 3421 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc 3481 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg 3541 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt 3601 ttatcggcac tttgcatccg ccgcgctccc gattccggaa gtgcttgaca ttggggagtt 3661 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct 3721 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggcaatgg atgccatcgc 3781 tgccgccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca 3841 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca 3901 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct 3961 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa 4021 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg 4081 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga 4141 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cccggctccg 4201 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt 4261 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac 4321 tgtcgggcgt acacaaatcg cccgcagaag cgccgccgtc tggaccgatg gctgtgtaga 4381 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaataagc 4441 tagtatgtaa gcctagtctt agataataaa atcgctatcc atcgaagatg gatgtgtgtt 4501 ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc gagaacgatc ctctcagtgc 4561 gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca gcttgggacc 4621 caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt accgatctgt 4681 ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta gcttttgcaa 4741 acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa catttccgtg 4801 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc 4861 tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac atcgaactgg 4921 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt ccaatgatga 4981 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc 5041 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca ccagtcacag 5101 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga 5161 gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag gagctaaccg 5221 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga 5281 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct 5341 tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag ttgatagact 5401 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt 5461 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg 5521 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta 5581 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac 5641 cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact 5701 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt 5761 gtcctcctta ccagaaattt atccttaaga tcccgaatcg tttaaactcg actctggctc 5821 tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg 5881 gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc ggctcccgac 5941 atcttggacc attagctcca caggtatctt cttccctcta gtggtcataa cagcagcttc 6001 agctacctct caattcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc 6061 tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca tccatcttcg atggatagcg 6121 attttattat ctaactgctg atcgagtgta gccagatcta gtaatcaatt acggggtcat 6181 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg 6241 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa 6301 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact 6361 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta 6421 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt 6481 acatctacgt attagtcatc gctattacca tgctgatgcg gttttggcag tacatcaatg 6541 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg 6601 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc 6661 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt 6721 tagtgaaccg tcagatcaga tctttgtcga tcctaccatc cactcgacac acccgccagc 6781 ggccgc //