LOCUS p1.1-Tr2-RBDv1 7575 bp DNA circular SYN 28-OCT-2020 DEFINITION synthetic plasmid p1.1-Tr2-RBDv1 for SARS-CoV-2 S-protein receptor binding domain expression. ACCESSION p1.1-Tr2-RBDv1 VERSION KEYWORDS . SOURCE synthetic construct ORGANISM synthetic construct other sequences; artificial sequences. REFERENCE 1 (bases 1 to 7575) AUTHORS Sinegubova,M.V., Orlova,N.A. and Vorobiev,I.I. TITLE High level expression of the monomeric SARS-CoV-2 S-protein RBD 320-537 in stably transfected CHO cells by the EEF1A1-based plasmid vector JOURNAL unpublished REFERENCE 2 (bases 1 to 7575) AUTHORS Sinegubova,M.V., Orlova,N.A. and Vorobiev,I.I. TITLE Direct Submission JOURNAL Submitted (28-OCT-2020) lab of Mammalian Cells Bioengineering, Institute of Bioengineering of The Federal Research Centre 'Fundamentals of Biotechnology' of the Russian Academy of Sciences, 60 let Oktjabrja pr-t, 7, bld. 1, Moscow, 117312, Russia COMMENT Bankit Comment: Vecscreen Comment:Submitter says that this sequence is a Synthetic Construct Bankit Comment: ALT EMAIL:ptichman@gmail.com Bankit Comment: TOTAL # OF SEQS:1. FEATURES Location/Qualifiers source 1..7575 /organism="synthetic construct" /mol_type="other DNA" /db_xref="taxon:32630" CDS <99..>140 /note="The native N-terminal signal peptide of SARS-CoV-2 S protein" /codon_start=1 /transl_table=11 /product="leader" /translation="MFVFLVLLPLVSSQ" CDS <144..>797 /note="SARS-CoV-2 S protein receptor binding domain (RBD)" /codon_start=1 /transl_table=11 /product="S protein RBD" /translation="VQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVA DYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADY NYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTP CNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNK " CDS <810..>839 /note="c-myc tag" /codon_start=1 /transl_table=11 /product="c-myc" /translation="EQKLISEEDL" CDS <855..875 /note="6xHis tag for IMAC purification" /codon_start=1 /transl_table=11 /product="6xHis" /translation="HHHHHH" CDS 1497..2060 /EC_number="1.5.1.3" /note="dihydrofolate reductase from Mus Musculus" /codon_start=1 /transl_table=11 /product="DHFR" /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSV EGKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIE QPELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLL PEYPGVLSEVQEEKGIKYKFEVYEKKD" regulatory 2062..2676 /regulatory_class="other" /note="downstream flanking region of CHO EEF1A1 gene" regulatory 2327..2333 /regulatory_class="polyA_signal_sequence" regulatory complement(2841..3514) /regulatory_class="replication_regulatory_region" CDS complement(3656..4516) /EC_number="3.5.2.6" /note="ampicillin resistance protein" /codon_start=1 /transl_table=11 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" regulatory complement(4511..4615) /regulatory_class="promoter" regulatory complement(4651..5052) /regulatory_class="other" /note="EBVTR" regulatory 5062..7573 /regulatory_class="other" /note="upstream flanking region of CHO EEF1A1 gene" regulatory 6555..6560 /regulatory_class="TATA_box" BASE COUNT 1776 a 2031 c 2048 g 1720 t ORIGIN 1 tcgaggccgc caccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct 61 ggttattgtg ctgtctcatc attttggcaa aggccaccat gttcgtgttt ctggtgctgc 121 tgcctctggt gtccagccag cgggtgcagc ccaccgaatc catcgtgcgg ttccccaata 181 tcaccaatct gtgccccttc ggcgaggtgt tcaatgccac cagattcgcc tctgtgtacg 241 cctggaaccg gaagcggatc agcaattgcg tggccgacta ctccgtgctg tacaactccg 301 ccagcttcag caccttcaag tgctacggcg tgtcccctac caagctgaac gacctgtgct 361 tcacaaacgt gtacgccgac agcttcgtga tccggggaga tgaagtgcgg cagattgccc 421 ctggacagac aggcaagatc gccgactaca actacaagct gcccgacgac ttcaccggct 481 gtgtgattgc ctggaacagc aacaacctgg actccaaagt cggcggcaac tacaattacc 541 tgtaccggct gttccggaag tccaatctga agcccttcga gcgggacatc tccaccgaga 601 tctatcaggc cggcagcacc ccttgtaacg gcgtggaagg cttcaactgc tacttcccac 661 tgcagtccta cggctttcag cccacaaatg gcgtgggcta tcagccctac agagtggtgg 721 tgctgagctt cgaactgctg catgcccctg ccacagtgtg cggccctaag aaaagcacca 781 atctcgtgaa gaacaaatgc gtgaacttcg aacaaaaact catctcagaa gaggatctga 841 atatgcatac cggtcatcat caccatcacc attaggctag cggccgcgca gttaacgccg 901 cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg 961 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg 1021 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg 1081 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca 1141 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct 1201 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca 1261 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa 1321 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggta 1381 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg 1441 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaagatct gccaccatgg 1501 ttcgaccatt gaactgcatc gtcgccgtgt cccaaaatat ggggattggc aagaacggag 1561 acctaccctg gcctccgctc aggaacgagt tcaagtactt ccaaagaatg accacaacct 1621 cttcagtgga aggtaaacag aatctggtga ttatgggtag gaaaacctgg ttctccattc 1681 ctgagaagaa tcgacctcta aaggacagaa ttaatatagt tctcagtaga gaactcaaag 1741 aaccaccacg aggagctcat tttcttgcca aaagtttgga tgatgcctta agacttattg 1801 aacaaccgga attggcaagt aaagtagaca tggtttggat agtcggaggc agttctgttt 1861 accaggaagc catgaatcaa ccaggccacc tcagactctt tgtgacaagg atcatgcagg 1921 aatttgaaag tgacacgttt ttcccagaaa ttgatttggg gaaatataaa cttctcccag 1981 aatacccagg cgtcctctct gaggtccagg aggaaaaagg catcaagtat aagtttgaag 2041 tctacgagaa gaaagactaa tcatattacc cctaacacct gccaccccag tcttaatcag 2101 tggtggaaga acggtctcag aactgtttgt ctcaattggc catttaagtt taatagtgaa 2161 agactggtta atgataacaa tgcatcggaa aaccttcagg aggaaaggag aatgttttgt 2221 ggaacatttt tgtgtgtgtg gcagttttaa gttattagtt ttcaaaatca gtacttttta 2281 atggaaacaa cttgaccaaa aatctgtcac agaattttga gacccattaa aatacaagtt 2341 taatgagaag tctgtctctg ttaatgctga agtcattact aagtgcttag cttagcaagg 2401 tatgtggatg cccatttgtg ttccaaggga ttggactgtt catcaggacc cagagctgag 2461 tttcaagggc tcaagagatg gcttattacc tgtgggtgtc ttgaaggttc tggttgggac 2521 aaattaggaa tgtttttggc agacatggtg actacccttc atctgggtga gttcagttga 2581 tttgtcttga gcctttgggg tttacacaag taaatgacat catacagtta gtgtattgtt 2641 agtgaatatt aatatatgag gcaggctttg ctctagctag acatatgtcg acgtccggag 2701 ctcaggcgcg ccatggctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 2761 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 2821 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 2881 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 2941 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 3001 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 3061 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 3121 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 3181 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 3241 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 3301 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg 3361 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 3421 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 3481 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 3541 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 3601 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc 3661 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 3721 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg 3781 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 3841 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta 3901 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 3961 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 4021 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 4081 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg 4141 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga 4201 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 4261 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 4321 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt 4381 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 4441 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 4501 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt 4561 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc 4621 gcacatttcc ccgaaaagtg ccacctgcat ctcatcacgg tcacgcatgg ctgcgcccgc 4681 gcagcgcccc ccgacccccc cccgatcccc ggaacgccag ccgccatcca ccgcccgcag 4741 cccccggagc ccgcggaccc cgaccccccg ccgctcgccc gccgttgcgc cgctctctgc 4801 gggggggcta gggggccgcg gggggaaggc cacgccccct ccactttttc caggaatgcg 4861 cggcatgacc cgcccccccc atgacccgcc aacacccccc cgatgccccc cgggggtctt 4921 tcctgggggg cctttgtcag ggttgcctgt gtcacccacg ggccccttgg ccacgcttgc 4981 ctgggtcacc ccggagcccc gggtcacgtt tgccggggtc agccctgggg cgcttcttca 5041 cgctccattg ccatgcagaa ttcccacatg tggacaccgg acagtaggtc ctcaaatgct 5101 ccttattagg ttggttcaat aatatcaatt gtttgttact aggcagtgat gttgtacatc 5161 tggaggagat ctcttgagcc cataatcagg ttattaggaa taaatactct aaggctaaaa 5221 atgtagctta gtgataagag tgcttgcctg gtgtgctgag accctcggtt ccatctccac 5281 aaccccatat tccattacaa aatacctttt caccgtccct agcattaaga aacaaaacaa 5341 caaagaagtt tttctttctt ctgagatcct gcccggagag gcatttaaaa ctggccaggg 5401 ccaaaaaaaa aaaaaaaaaa agaaaaaaaa gaaaagaaaa caggctaggg ccggcatggt 5461 ggcgcacgcc tttaatccca gcacgcagga ggcagaggca gggcggatct ctgtgagttt 5521 gaggtcagcc tggtctacct agtgagtttc agggcaccca gggctaaaga gactgtctca 5581 aaaacaaaac agccacacaa tcagaaccac agcaaaacgc agttatgatc cttggaactg 5641 taggaatgac aagcatttaa ataataggac gagccatttt tgagaagctc tgatttcaca 5701 agtgtcaggg atgggctctg ggcgagtaag attgctaatg ctggcctcta aatgagacca 5761 cgtggagttg attagattct tttcatgttc ctcgtgctct atcaaataac tgtacccaaa 5821 tacacacaca cacacacaca cacaatgcgc gcacacacaa aatccttttt tagcttaaga 5881 agcccagaat cagaagtaaa gctaactgtg ggacttaagt attattctga acggaactcc 5941 cagggcgtga agcgcgcttc aggcttccag agaagcagct ggcgctggat ggaatgaacc 6001 aagaggccag cacaggggca gatccgtcga gctctcggcc accgagctga gcccttaggt 6061 tctggggctg ggaagggtcc ctaggattgt gcacctctcc cgcgggggac aagcaggggg 6121 atggcggggc tgacgtcggg aggtggcctc cacgggaagg gacacccgga tctcgacaca 6181 gccttggcag tggagtcagg aagggtaggg acagattctg gacgccctct tggccagctc 6241 ctcaccgccc cacccccgac tggagccgag agtaattcat acaaaaggag ggatcgcctt 6301 cgcccctggg aatcccaggg accgtcgcta aattctggcc ggcctcccag cccggaaccg 6361 ctgtgcccgc ccagcgcggc gggaggagcc tgcgcctagg gcggatcgcg ggtcggcggg 6421 agagcacaag cccacagtcc ccggcggtgg gggaggggcg cgctgagcgg gggcccggga 6481 gccagcgcgg ggcaaactgg gaaagtggtg tcgtgtgctg gctccgccct cttcccgagg 6541 gtgggggaga acggtataaa agtgcggtag tcgcgttgga cgttcttttt cgcaacgggt 6601 ttgccgtcag aacgcaggtg agtggcgggt gtggcctccg cgggcccggg ctccctcctt 6661 tgagcggggt cggaccgccg tgcgggtgtc gtcggccggg cttctctgcg agcgttcccg 6721 ccctggatgg cgggctgtgc gggagggcga gggggggagg cctggcggcg gccccggagc 6781 ctcgcctcgt gtcgggcgtg aggcctagcg tggcttccgc cccgccgcgt gccaccgcgg 6841 ccgcgctttg ctgtctgccc ggctgccctc gattgcctgc ccgcggcccg ggccaacaaa 6901 gggagggcgt ggagctggct ggtagggagc cccgtagtcc gcatgtcggg cagggagagc 6961 ggcagcagtc ggggggggga ccgggcccgc ccgtcccgca gcacatgtcc gacgccgcct 7021 ggacgggtag cggcctgtgt cctgataagg cggccgggcg gtgggtttta gatgccgggt 7081 tcaggtggcc ccgggtcccg gcccggtctg gccagtaccc cgtagtggct tagctccgag 7141 gagggcgagg cccgcccgcc cggcaccagt tgcgtgcgcg gaaagatggc cgctcccggg 7201 ccctgtagca aggagctcaa aatggaggac gcggcagccc ggcggagcgg ggcgggtgag 7261 tcacccacac aaaggaagag ggccttgccc ctcgccggcc gctgcttcct gtgaccccgt 7321 ggtgtaccgg ccgcacttca gtcaccccgg gcgctctttc ggagcaccgc tggcctccgc 7381 tgggggaggg gatctgtcta atggcgttgg agtttgctca catttggtgg gtggagactg 7441 tagccaggcc agcctggcca tgaaagtaat tcttggaatt tgcccatttt gagtttggag 7501 cgaagctgat tgacaaagct gcttagccgt tcaaaggtat tcttcgaact ttttttttaa 7561 ggtgttgtga aaacc //