LOCUS Exported 9215 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Express SARS-CoV spike protein with C9 tag at C-terminal in mammalian cells. ACCESSION . VERSION . KEYWORDS pcDNA3.1-SARS-Spike SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9215) AUTHORS Shang J, Ye G, Shi K, Wan Y, Luo C, Aihara H, Geng Q, Auerbach A, Li F TITLE Structural basis of receptor recognition by SARS-CoV-2. JOURNAL Nature. 2020 Mar 30. pii: 10.1038/s41586-020-2179-y. doi: 10.1038/s41586-020-2179-y. PUBMED 32225175 REFERENCE 2 (bases 1 to 9215) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9215 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 149..528 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 529..732 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 683..703 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 777..796 /label=T7 /note="T7 promoter, forward primer" promoter 777..795 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4722..4739) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4728..4952 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4998..5426 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5085..5104) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5295..5316 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5435..5455) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5440..5769 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5620..5755 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5682..5701 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(5890..5909) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6498..6517 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6802..6923 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6839..6858) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6893..6912 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6972..6988) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6972..6988) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6985..7007) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6996..7012 /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(7020..7050) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7065..7086 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7203..7220) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7374..7962) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7454..7473) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8133..8993) /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 8756..8775 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8994..9098) /gene="bla" /label=AmpR promoter primer_bind complement(9173..9192) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg 61 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt 121 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa 181 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg 241 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg 301 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta 361 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt 421 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac 481 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt 541 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac 601 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt 661 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat 721 ataagcagag ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat 781 acgactcact atagggagac ccaagctggc tagcgtttaa acgggccctc tagaaaccat 841 gcccatgggg tctctgcaac cgctggccac cttgtacctg ctggggatgc tggtcgcttc 901 cgtgctagcc agcggcagcg acctggaccg ctgcaccacg ttcgatgacg tgcaggcgcc 961 caactacacc cagcacacct ccagcatgcg cggcgtgtac tatcccgacg agatcttccg 1021 cagcgacacc ctgtacctca cccaggacct gttcctgccc ttctactcca acgtgaccgg 1081 cttccacacc atcaaccaca ccttcggcaa ccccgtgatc ccgttcaagg acggcatcta 1141 cttcgccgcg accgagaaga gcaacgtggt ccgcggctgg gtgttcggga gcaccatgaa 1201 taacaagtcc cagagcgtga tcattatcaa taactccacc aacgtcgtga tccgcgcctg 1261 caacttcgag ctgtgtgaca acccgttctt tgccgtgagc aagcccatgg gcacccagac 1321 gcacaccatg atcttcgaca acgccttcaa ctgcaccttc gagtacatca gcgacgcctt 1381 ctccctggac gtgagcgaga agtccggaaa cttcaagcac ctgcgcgagt tcgtgttcaa 1441 gaacaaagac ggcttcctgt acgtgtacaa gggctaccag cccatcgacg tcgtgcgcga 1501 cctgcccagc ggcttcaaca ccctgaagcc catcttcaag ctgcccctcg gcattaacat 1561 caccaacttc cgcgccatcc tgaccgcctt cagccctgcc caggacatct ggggcaccag 1621 cgccgcggcc tacttcgtcg gctacctgaa gcccacgacc ttcatgctga agtacgacga 1681 gaacggcacg atcaccgatg ccgtggactg cagccagaac ccgctcgccg agctgaagtg 1741 cagcgtgaag tccttcgaga tcgacaaggg catctaccag accagcaact tccgcgtggt 1801 acccagcggc gacgtggtcc gcttcccgaa catcaccaac ctgtgtccct tcggcgaggt 1861 gttcaacgcc accaagttcc cgagcgtgta cgcctgggag cgcaagaaga tcagcaactg 1921 cgtggccgac tacagcgtgc tgtacaactc caccttcttt agcaccttca agtgctacgg 1981 cgtgagcgcc accaagctca acgacctgtg cttcagcaac gtgtacgccg acagcttcgt 2041 ggtcaagggc gacgatgtgc gccagatcgc gcccggccag accggcgtga tcgccgacta 2101 caactacaag ctgcccgatg acttcatggg ctgcgtgctg gcctggaaca cccgcaacat 2161 cgacgccacg agcaccggca actacaacta caagtatcgc tacctgcgcc acggcaagct 2221 gcggcccttc gagcgcgaca tctccaacgt gcccttcagc ccggacggca agccctgcac 2281 gccgcccgcc ctgaactgct actggccgct gaacgactat ggcttctaca ctacgaccgg 2341 tatcggctac cagccctacc gcgtggtcgt gctgagcttc gagctcctga acgcgcccgc 2401 caccgtgtgc ggcccgaagc tgagcaccga cctgatcaag aaccagtgcg tgaacttcaa 2461 tttcaacggc ctgaccggca ccggcgtgct gacgcccagc tccaagcgct tccagccctt 2521 ccagcagttc ggccgcgacg tcagcgactt caccgacagc gtgcgcgacc cgaagaccag 2581 cgagatcctg gacatctctc cctgctcctt cggcggtgtc agcgtgatca cgcccggcac 2641 caacgccagc tccgaggtgg ccgtcctgta ccaggacgtg aactgcaccg acgtcagcac 2701 cgccatccac gcggaccagc tgacgcccgc ctggcgcatc tacagcaccg gcaacaacgt 2761 gttccagacc caggccggct gcctgatcgg cgccgagcac gtcgacacca gctacgagtg 2821 cgacatcccg attggcgccg ggatctgcgc cagctaccac accgtgagcc tgctccgctc 2881 caccagccag aagtccatcg tggcctacac catgagcctg ggcgccgaca gcagcatcgc 2941 ctacagcaac aacaccatcg ccatcccgac caacttctcc atcagcatca cgaccgaggt 3001 gatgcccgtg agcatggcca agaccagcgt ggactgcaac atgtacatct gcggcgacag 3061 caccgagtgc gccaacctgc tcctgcagta cggctccttc tgcacccagc tgaaccgcgc 3121 cctgagcggc atcgccgccg agcaggaccg caacacccgc gaggtgttcg cccaggtgaa 3181 gcagatgtac aagacgccca ccctgaagta cttcggcggc ttcaacttca gccagatcct 3241 gcccgacccg ctgaagccca ccaagcgcag cttcatcgag gacctgctct tcaacaaggt 3301 gacgctggcc gacgccggct tcatgaagca gtacggcgag tgcctgggcg acatcaacgc 3361 ccgcgacctg atctgcgccc agaagttcaa cggcctgacc gtgctcccgc cgctgctgac 3421 cgacgacatg atcgccgcgt acaccgccgc gctggtgagc ggcaccgcca cggccggctg 3481 gaccttcggc gccggcgcgg ccctgcagat cccgttcgcc atgcagatgg cctaccgctt 3541 caacggcatc ggcgtgaccc agaacgtgct gtacgagaac cagaagcaga tcgccaacca 3601 gttcaacaag gccatcagcc agatccagga gagcctgacc acgaccagca cggccctggg 3661 caagctgcag gacgtggtca accagaacgc ccaggccctg aacaccctgg tgaagcagct 3721 gtccagcaac ttcggcgcca tcagctccgt gctgaacgat atcctgagcc gcctggacaa 3781 ggtggaggcc gaggtgcaga tcgatcgcct gatcaccggc cgcctgcaga gcctgcagac 3841 ctacgtgacc cagcagctga tccgcgccgc ggagatccgc gccagcgcga acctggccgc 3901 caccaagatg tccgagtgcg tgctgggcca gagcaagcgc gtggacttct gcggcaaggg 3961 ctaccacctg atgagcttcc cgcaggccgc gccgcacggc gtggtcttcc tgcacgtgac 4021 ctacgtgccc agccaggagc gcaacttcac cacggcgccc gccatctgcc acgagggcaa 4081 ggcctacttc ccgcgcgagg gcgtgttcgt gttcaacggc accagctggt tcatcaccca 4141 gcgcaacttc ttcagcccgc aaatcatcac cacggacaac accttcgtga gcggcaactg 4201 cgacgtcgtg atcggcatca tcaacaacac cgtgtacgac ccgctgcagc ccgagctgga 4261 ctccttcaag gaggagctgg acaagtactt caagaaccac accagcccgg acgtggacct 4321 gggcgacatc agcggcatca acgccagcgt cgtgaacatc cagaaggaga tcgaccgcct 4381 gaacgaggtg gccaagaacc tgaacgagag cctgatcgac ctccaggagc tgggcaagta 4441 cgagcagtac atcaagtggc cttggtatgt gtggctgggc ttcatcgccg ggctgatcgc 4501 catcgtgatg gtgaccatcc tgctctgctg catgaccagc tgctgctcct gcctgaaggg 4561 cgcctgcagc tgcggcagct gctgcaagtt cgacgaggac gacagcgagc ccgtgctgaa 4621 gggcgtgaag ctgcactaca cgggcggcac cgagaccagc caggtggcgc ccgcctagga 4681 tccgagctcg gtaccaagct taagtttaaa ccgctgatca gcctcgactg tgccttctag 4741 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac 4801 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca 4861 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag 4921 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg 4981 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt 5041 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt 5101 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc 5161 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga 5221 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc 5281 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt 5341 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct 5401 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga 5461 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 5521 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 5581 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc 5641 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 5701 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc 5761 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga 5821 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg 5881 gtggagaggc tattcggcta tgactgggca acagacaatc ggctgctctg atgccgccgt 5941 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc 6001 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc 6061 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga 6121 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat 6181 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca 6241 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga 6301 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc 6361 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat 6421 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga 6481 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg 6541 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt 6601 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa 6661 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg 6721 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 6781 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc 6841 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 6901 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg 6961 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 7021 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 7081 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 7141 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 7201 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 7261 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 7321 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 7381 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 7441 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 7501 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 7561 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 7621 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 7681 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 7741 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 7801 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 7861 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 7921 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 7981 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 8041 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 8101 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 8161 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 8221 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 8281 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 8341 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 8401 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 8461 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 8521 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 8581 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 8641 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 8701 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 8761 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 8821 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 8881 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 8941 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 9001 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 9061 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 9121 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct 9181 gctctgatgc cgcatagtta agccagtatc tgctc //