LOCUS Exported 10023 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Express NSP9 gene along with HA and BASU. ACCESSION . VERSION . KEYWORDS SARS_COV2_nsp9_HA_BASU SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10023) TITLE Khavari lab SARS-CoV-2 plasmids REFERENCE 2 (bases 1 to 10023) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10023 /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)" promoter 2338..3519 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2568..3510 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3467..3487 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 3910..3933 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 3937..3963 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" misc_feature 4773..5346 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4940..4957) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5167..5186 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5379..5978 /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(5379..5398) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 5875..5895 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 5992..6580 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(6045..6065) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6463..6474) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 6787..7420 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" primer_bind complement(7548..7564) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7548..7564) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7561..7583) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7572..7588 /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(7596..7626) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7641..7662 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7779..7796) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7950..8538) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8030..8049) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8709..9569) /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 9332..9351 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9570..9674) /gene="bla" /label=AmpR promoter polyA_signal 9722..9856 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(9759..9778) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 9813..9832 /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 cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg 2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc 2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct 2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag 2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg 2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag 2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt 2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt 2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct 3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gatctatttc cggtgaattc 3541 gccgccacca tgaataatga gctttctccg gttgctctgc gacagatgtc ctgcgcggct 3601 ggaacaacac aaactgcctg tactgacgac aacgcactgg cgtactataa taccaccaaa 3661 ggaggtcgat ttgtattggc acttctgtcc gacttgcagg accttaaatg ggcgaggttt 3721 cccaagagtg acggaacggg cacaatttac acggagctgg agcccccatg ccggtttgtg 3781 actgatactc caaaaggtcc taaagtaaaa tacttgtatt ttattaaagg tttgaataat 3841 ttgaacaggg gtatggtact cggctccctg gctgcgaccg tgcgactcca actcgaaggc 3901 ggcgggggat ggagccatcc acaattcgag aaaggctacc catacgatgt gcctgactat 3961 gctggcaagt tgtctgaatc agagattagg tttggactga aaacggaggt gatgggccaa 4021 catcttatct atcacgatgt tttgtccagc acacaaaaga cagcccatga attggcaaat 4081 aacaatgccc ccgagggtac cctcgtggtt gcagataaac agacagccgg taggggcggg 4141 atgagccgag tgtggcattc tcaagaagga aacggagtat ggatgagttt gatattgcgc 4201 ccggacattc cgctccaaaa gacgccccaa cttaccttgc tggcagctgt agcggtggtt 4261 cagggaatag aagaggccgc gggcatacag actgatataa agtggccgaa cgacatactc 4321 attaacggga aaaaaacagt agggatactt accgaaatgc aagctgagga ggatcgagtg 4381 cgctccgtga ttattggtat tggcattaat gtaaatcagc aacccaatga tttccccgat 4441 gagctcaagg acattgcgac gagtctgtcc caggcagctg gggaaaagat cgacagggca 4501 ggtgtaatcc agcatatctt gctttgtttc gaaaagcgat accgagacta catgacccac 4561 ggtttcacgc ccatcaagct cttgtgggag tcatacgccc tcggtattgg cacgaacatg 4621 agggcgagaa ctctcaacgg gacattttac ggtaaggcgt tgggtattga cgacgaaggg 4681 gtacttttgt tggaaactaa tgaaggcatt aagaagatat acagcgcgga catcagcctt 4741 aggtgaccgc agtttgaaaa gtaaggatcc cgcccctctc cctccccccc ccctaacgtt 4801 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc 4861 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc 4921 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag 4981 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg 5041 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat 5101 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga 5161 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc 5221 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg 5281 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga 5341 tgataagctt gccacaaccc acaaggagac gaccttccat gaccgagtac aagcccacgg 5401 tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg 5461 ccgactaccc cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg 5521 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg 5581 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt 5641 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac 5701 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg 5761 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag 5821 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc 5881 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac 5941 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctagac gcgtctggaa caatcaacct 6001 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 6061 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 6121 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 6181 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 6241 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 6301 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 6361 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt 6421 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 6481 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 6541 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattaa ttctgcagtc 6601 gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt 6661 gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac 6721 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaaga 6781 ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct 6841 accacacaca aggctacttc cctgattagc agaactacac accagggcca ggggtcagat 6901 atccactgac ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag 6961 aggccaataa aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg 7021 acccggagag agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg 7081 cccgagagct gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac 7141 tttccgctgg ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc 7201 tcagatcctg catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag 7261 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc 7321 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga 7381 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt 7441 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gccttgacat 7501 tgctagcgtt taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt 7561 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 7621 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 7681 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 7741 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 7801 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 7861 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 7921 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 7981 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 8041 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 8101 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 8161 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 8221 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 8281 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 8341 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 8401 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 8461 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 8521 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 8581 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 8641 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 8701 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 8761 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 8821 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 8881 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 8941 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 9001 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 9061 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 9121 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 9181 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 9241 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 9301 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 9361 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 9421 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 9481 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 9541 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 9601 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 9661 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat 9721 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 9781 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 9841 ttatcatgtc tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac 9901 cccataccct attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc 9961 ctctgaatta ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt 10021 agt //