LOCUS Exported 9225 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses SARS-CoV-2 spike protein. ACCESSION . VERSION . KEYWORDS pcDNA3.1 SARS-CoV-2 S D614 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9225) AUTHORS Yurkovetskiy L, Wang X, Pascal KE, Tomkins-Tinch C, Nyalile TP, Wang Y, Baum A, Diehl WE, Dauphin A, Carbone C, Veinotte K, Egri SB, Schaffner SF, Lemieux JE, Munro JB, Rafique A, Barve A, Sabeti PC, Kyratsous CA, Dudkina NV, Shen K, Luban J TITLE Structural and Functional Analysis of the D614G SARS-CoV-2 Spike Protein Variant. JOURNAL Cell. 2020 Sep 15. pii: S0092-8674(20)31229-0. doi: 10.1016/j.cell.2020.09.032. PUBMED 32991842 REFERENCE 2 (bases 1 to 9225) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9225 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4816..4833) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4822..5046 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5092..5520 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5179..5198) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5389..5410 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5529..5549) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5534..5863 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5714..5849 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5776..5795 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5930..6724 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5984..6003) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6594..6613 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6898..7019 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6935..6954) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6989..7008 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(7068..7084) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7068..7084) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7081..7103) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7092..7108 /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(7116..7146) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7161..7182 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7299..7316) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7470..8058) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7550..7569) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8229..9089) /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 8852..8871 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9090..9194) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaacgg gccctctaga gccaccatgt ttgtgttctt ggtcttgttg cctttggttt 961 caagtcagtg cgtgaatttg actacacgaa ctcagctgcc gcctgcctac accaattcct 1021 tcacgcgagg ggtctactac ccagataaag tttttcggag tagtgttctc cactctaccc 1081 aggacctttt cctccccttc tttagcaacg tcacatggtt ccatgcaata cacgttagcg 1141 ggacgaatgg gaccaaacgc ttcgataatc cggtactccc gttcaatgat ggggtctact 1201 tcgcttctac tgagaagtca aacattatcc gcggttggat attcggcaca acacttgatt 1261 ctaaaactca gtctctcctg attgtgaata atgctaccaa tgttgtgatt aaggtctgcg 1321 agttccaatt ttgcaatgac ccgttcctgg gcgtttatta tcataagaat aacaagagct 1381 ggatggagtc tgagttccgc gtatactctt ctgccaataa ttgtaccttc gaatacgtca 1441 gtcagccgtt cttgatggac ctcgaaggaa agcaaggcaa tttcaaaaac ctcagggagt 1501 tcgtttttaa gaatatcgat ggttacttta agatatattc aaaacataca cccataaacc 1561 ttgtccgaga tctgccgcaa ggatttagtg cacttgaacc gctggtcgat cttcctattg 1621 gaattaacat tacccggttt caaactctgt tggctttgca ccggagttat cttacgccag 1681 gcgacagctc cagtggatgg acggcagggg ctgctgcata ctatgtcggc taccttcaac 1741 cccggacttt tctcctcaag tacaacgaga atgggaccat aacagacgct gtagattgtg 1801 ctctggatcc ccttagcgaa acaaagtgca cgctcaagtc ctttacggtt gaaaaaggga 1861 tctatcagac cagtaatttc cgcgtacaac ccaccgaaag cattgtacgg tttccgaaca 1921 ttaccaatct gtgtcctttc ggtgaggttt tcaatgcgac ccggttcgca tcagtctacg 1981 catggaaccg caagcgcatt agcaattgtg tcgccgatta tagcgtgctt tacaattctg 2041 cctctttctc cacattcaag tgttatggtg tgtcacctac taagctcaat gacctgtgtt 2101 ttacaaatgt ttacgccgac agctttgtca ttaggggcga tgaggtccgc cagattgcgc 2161 ctgggcagac gggaaaaata gccgattata actataagct tcctgatgat tttaccggct 2221 gtgtgatagc ctggaactca aacaatcttg attctaaagt gggaggaaac tataactatc 2281 tctaccggct ctttcgcaag tccaacctca aaccgttcga gcgagacata agcacagaaa 2341 tctaccaagc aggtagtacg ccgtgcaacg gggtagaagg gtttaattgt tacttcccgt 2401 tgcagagtta cggatttcaa ccaaccaacg gagtagggta tcagccgtac cgcgttgtgg 2461 ttctctcatt cgaacttctg catgcacccg cgacagtatg tggtccaaag aagtctacga 2521 acctcgttaa gaataaatgc gtgaacttta actttaatgg cctgacaggg acgggggtcc 2581 ttaccgagag caacaagaaa tttttgccgt tccagcagtt tgggagggat attgctgaca 2641 ctaccgacgc agttcgcgac ccacagactc tggaaatact tgacatcaca ccttgctcat 2701 ttggaggggt ctccgtcata acccctggta ctaacacatc taatcaagtt gctgttctct 2761 atcaggatgt aaattgtaca gaggttcctg tagccatcca tgctgaccag ctgacaccca 2821 cctggagggt ttatagtacc ggaagcaacg ttttccaaac tagagcaggt tgtttgattg 2881 gtgctgagca cgtaaataac tcatacgaat gcgacatacc tattggagcg ggtatctgtg 2941 caagctacca aactcaaacc aattcaccga ggcgcgcacg atccgttgcg agccaatcca 3001 taattgccta cacgatgagc ctcggcgcag aaaactcagt tgcgtattcc aataacagca 3061 ttgcaatccc tacaaatttt acgatcagtg ttacgaccga aatcctgccg gtatcaatga 3121 cgaaaacaag cgtggactgc accatgtaca tctgtgggga tagtaccgag tgtagtaact 3181 tgctgctcca gtatgggtca ttttgcactc aactcaacag agctttgaca ggcatcgccg 3241 ttgaacaaga taaaaatacc caagaagtat ttgctcaagt caaacaaatt tacaagactc 3301 ctcccataaa agactttgga gggttcaact tttcccagat attgccggac ccatcaaagc 3361 caagcaagcg gtcatttatt gaagacttgc tcttcaacaa agttacactt gctgacgcgg 3421 gattcattaa acagtatggt gactgtttgg gagatattgc ggcgcgggac ctgatatgtg 3481 ctcaaaagtt caatgggctg acggtgctgc ccccgctgct tacggacgaa atgattgccc 3541 aatacacatc tgctctcctt gccggtacca ttacttcagg gtggacgttc ggcgcaggag 3601 ctgcccttca aatacctttt gcaatgcaaa tggcatatcg gttcaacgga ataggtgtca 3661 cgcagaacgt attgtacgag aaccaaaagc tcattgcaaa tcaatttaac tcagcgatcg 3721 gtaagataca ggactcactt agctctaccg cttccgccct ggggaagctg caagatgtcg 3781 taaaccagaa cgctcaagcg cttaatacat tggttaaaca attatctagc aattttggtg 3841 ctatcagttc cgttcttaac gacattttgt ctcggcttga caaggttgag gcggaagttc 3901 aaatcgacag actcataaca ggacggctgc agtccctgca aacgtatgtc actcaacagc 3961 tcattcgggc cgcggagatt cgggcaagcg caaaccttgc ggccacaaaa atgagtgaat 4021 gtgtacttgg ccagtccaag agagttgatt tctgtggaaa aggataccac ctgatgtcct 4081 tcccgcaatc tgcgccgcac ggcgttgttt ttttgcatgt aacttatgta cctgcgcaag 4141 aaaaaaattt cacgactgcg cctgcaatat gtcatgacgg gaaggctcac ttcccaaggg 4201 aaggcgtctt tgtatcaaat gggacgcatt ggttcgtcac gcagcgcaac ttttatgagc 4261 cccagattat aactaccgac aacacattcg tcagcggcaa ttgtgatgtg gtaatcggca 4321 tagtcaacaa taccgtatac gatcctttgc aacctgaact tgactcattc aaggaggaat 4381 tggataagta tttcaaaaat catacctctc cagacgtcga tctcggtgat atatcaggta 4441 taaatgcgtc agtcgtaaac atccagaagg agattgatcg cttgaacgag gttgctaaga 4501 atttgaatga aagcctcatc gacctccagg aattagggaa atatgaacaa tacattaagt 4561 ggccttggta catctggttg gggttcatcg caggactcat cgcgatcgtg atggttacca 4621 taatgctttg ttgcatgacg agctgttgtt cctgtctgaa aggttgctgc tcttgcggca 4681 gttgctgcaa gtttgatgag gacgattctg aaccagttct caaaggggta aaattgcatt 4741 acacataaga attccaccac actggactag tggatccgag ctcggtacca agcttaagtt 4801 taaaccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct 4861 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg 4921 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc 4981 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct 5041 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct 5101 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg 5161 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg 5221 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac 5281 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct 5341 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt 5401 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt 5461 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt 5521 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag 5581 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc 5641 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc 5701 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg 5761 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca 5821 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg 5881 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca 5941 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg 6001 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg 6061 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc 6121 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt 6181 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc 6241 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca 6301 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc 6361 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg 6421 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct 6481 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc 6541 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc 6601 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta 6661 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt 6721 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga 6781 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac 6841 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac 6901 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 6961 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 7021 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt 7081 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 7141 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 7201 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 7261 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 7321 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 7381 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 7441 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 7501 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 7561 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 7621 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 7681 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 7741 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 7801 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 7861 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 7921 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 7981 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 8041 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 8101 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 8161 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 8221 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 8281 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 8341 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 8401 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 8461 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 8521 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 8581 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 8641 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 8701 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 8761 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 8821 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 8881 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 8941 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 9001 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 9061 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 9121 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 9181 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc //