LOCUS Exported 6036 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 helicase. ACCESSION . VERSION . KEYWORDS pGBW-m4134333 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6036) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6036) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6036 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 1805..1825 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1826..1852 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 1905..2026 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1916..1935) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1970..1989 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 2141..2187 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 2217..2353 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 2398..2469 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(2695..3283) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2775..2794) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 4029..4889 /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(4247..4266) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 4902..5038 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 5190..5261 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 5266..5312 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 5330..5378 /note="synthetic polyadenylation signal" enhancer 5459..5762 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 5764..5966 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5917..5937 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 6035..8 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" ORIGIN 1 caccatggtt ggggcttgtg ttctttgcaa ttcacagact tcattaagat gtggtgcttg 61 catacgtaga ccattcttat gttgtaaatg ctgttacgac catgtcatat caacatcaca 121 taaattagtc ttgtctgtta atccgtatgt ttgcaatgct ccaggttgtg atgtcacaga 181 tgtgactcaa ctttacttag gaggtatgag ctattattgt aaatcacata aaccacccat 241 tagttttcca ttgtgtgcta atggacaagt ttttggttta tataaaaata catgtgttgg 301 tagcgataat gttactgact ttaatgcaat tgcaacatgt gactggacaa atgctggtga 361 ttacatttta gctaacacct gtactgaaag actcaagctt tttgcagcag aaacgctcaa 421 agctactgag gagacattta aactgtctta tggtattgct actgtacgtg aagtgctgtc 481 tgacagagaa ttacatcttt catgggaagt tggtaaacct agaccaccac ttaaccgaaa 541 ttatgtcttt actggttatc gtgtaactaa aaacagtaaa gtacaaatag gagagtacac 601 ctttgaaaaa ggtgactatg gtgatgctgt tgtttaccga ggtacaacaa cttacaaatt 661 aaatgttggt gattattttg tgctgacatc acatacagta atgccattaa gtgcacctac 721 actagtgcca caagagcact atgttagaat tactggctta tacccaacac tcaatatctc 781 agatgagttt tctagcaatg ttgcaaatta tcaaaaggtt ggtatgcaaa agtattctac 841 actccaggga ccacctggta ctggtaagag tcattttgct attggcctag ctctctacta 901 cccttctgct cgcatagtgt atacagcttg ctctcatgcc gctgttgatg cactatgtga 961 gaaggcatta aaatatttgc ctatagataa atgtagtaga attatacctg cacgtgctcg 1021 tgtagagtgt tttgataaat tcaaagtgaa ttcaacatta gaacagtatg tcttttgtac 1081 tgtaaatgca ttgcctgaga cgacagcaga tatagttgtc tttgatgaaa tttcaatggc 1141 cacaaattat gatttgagtg ttgtcaatgc cagattacgt gctaagcact atgtgtacat 1201 tggcgaccct gctcaattac ctgcaccacg cacattgcta actaagggca cactagaacc 1261 agaatatttc aattcagtgt gtagacttat gaaaactata ggtccagaca tgttcctcgg 1321 aacttgtcgg cgttgtcctg ctgaaattgt tgacactgtg agtgctttgg tttatgataa 1381 taagcttaaa gcacataaag acaaatcagc tcaatgcttt aaaatgtttt ataagggtgt 1441 tatcacgcat gatgtttcat ctgcaattaa caggccacaa ataggcgtgg taagagaatt 1501 ccttacacgt aaccctgctt ggagaaaagc tgtctttatt tcaccttata attcacagaa 1561 tgctgtagcc tcaaagattt tgggactacc aactcaaact gttgattcat cacagggctc 1621 agaatatgac tatgtcatat tcactcaaac cactgaaaca gctcactctt gtaatgtaaa 1681 cagatttaat gttgctatta ccagagcaaa agtaggcata ctttgcataa tgtctgatag 1741 agacctttat gacaagttgc aatttacaag tcttgaaatt ccacgtagga atgtggcaac 1801 tttagaaaac ctttactttc agtcgtaccc ctatgacgta ccagattatg cgtaagctag 1861 cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat acattgatga 1921 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga 1981 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg 2041 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa 2101 cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc 2161 tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat 2221 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga 2281 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga 2341 aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat 2401 tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt 2461 ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag 2521 ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt 2581 tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg 2641 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 2701 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 2761 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 2821 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 2881 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 2941 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 3001 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 3061 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 3121 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 3181 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 3241 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 3301 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 3361 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 3421 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 3481 tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc 3541 aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc 3601 gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc 3661 tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc 3721 aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc 3781 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc 3841 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc 3901 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt 3961 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag 4021 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt 4081 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt 4141 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt 4201 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg 4261 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga 4321 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa 4381 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga 4441 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa 4501 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca 4561 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta 4621 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac 4681 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc 4741 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag 4801 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga 4861 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa 4921 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac 4981 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga 5041 tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc 5101 gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt 5161 acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt 5221 cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa 5281 gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa 5341 ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta 5401 gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 5461 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 5521 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 5581 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 5641 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 5701 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 5761 tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat 5821 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 5881 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac 5941 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga 6001 tcctaccatc cactcgacac acccgccagc ggccgc //