LOCUS Exported 8316 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression vector for expressing SARS-2 Spike with 21 aa del at C-terminus and pt mutation D to G at site 614. ACCESSION . VERSION . KEYWORDS HDM_SARS2_Spike_del21_D614G SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8316) AUTHORS Crawford KHD, Eguia R, Dingens AS, Loes AN, Malone KD, Wolf CR, Chu HY, Tortorici MA, Veesler D, Murphy M, Pettie D, King NP, Balazs AB, Bloom JD TITLE Protocol and Reagents for Pseudotyping Lentiviral Particles with SARS-CoV-2 Spike Protein for Neutralization Assays. JOURNAL Viruses. 2020 May 6;12(5). pii: v12050513. doi: 10.3390/v12050513. PUBMED 32384820 REFERENCE 2 (bases 1 to 8316) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8316 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 82..461 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 462..665 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 616..636 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 662..686 /label=LNCX /note="Human CMV promoter, forward primer" intron 779..1254 /label=-beta-globin intron /note="internally truncated intron from human beta-globin" primer_bind complement(5086..5102) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5087..5103) /label=pBluescriptKS /note="For pBluescript vector" polyA_signal 5196..5590 /label=-beta-globin poly(A) /note="human beta-globin polyadenylation signal" primer_bind complement(5924..5942) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6010..6114 /gene="bla" /label=AmpR promoter CDS 6115..6975 /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(6333..6352) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7146..7734 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7635..7654 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind complement(8277..8296) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" ORIGIN 1 agcttggccc attgcatacg ttgtatccat atcataatat gtacatttat attggctcat 61 gtccaacatt accgccatgt tgacattgat tattgactag ttattaatag taatcaatta 121 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 181 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 241 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 301 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 361 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 421 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 481 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 541 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 601 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 661 gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc 721 atagaagaca ccgggaccga tccagcctcc cctcgaagct gatcctgaga acttcagggt 781 gagtctatgg gacccttgat gttttctttc cccttctttt ctatggttaa gttcatgtca 841 taggaagggg agaagtaaca gggtacacat attgaccaaa tcagggtaat tttgcatttg 901 taattttaaa aaatgctttc ttcttttaat atactttttt gtttatctta tttctaatac 961 tttccctaat ctctttcttt cagggcaata atgatacaat gtatcatgcc tctttgcacc 1021 attctaaaga ataacagtga taatttctgg gttaaggcaa tagcaatatt tctgcatata 1081 aatatttctg catataaatt gtaactgatg taagaggttt catattgcta atagcagcta 1141 caatccagct accattctgc ttttatttta tggttgggat aaggctggat tattctgagt 1201 ccaagctagg cccttttgct aatcatgttc atacctctta tcttcctccc acagctcctg 1261 ggcaacgtgc tggtctgtgt gctggcccat cactttggca aagaattccg ccaccatgtt 1321 cgtgttcctg gtactccttc ctttggtgtc ttcccagtgt gtaaatctta ccactcggac 1381 ccagcttcca cccgcctaca ccaacagttt cacacgcggc gtctactatc ctgacaaggt 1441 gtttaggagt tcagtcttgc actcaactca agacttgttc ctccctttct ttagcaatgt 1501 gacgtggttt catgccattc atgtctccgg cacaaacgga acgaagcgct ttgataatcc 1561 tgtgctcccg ttcaacgatg gagtgtactt cgcgtccaca gagaagagca atatcattcg 1621 aggttggatc ttcggaacga cactcgactc aaagacgcag tcccttctca tcgtcaataa 1681 tgccacgaac gtggtcatca aagtgtgcga gtttcaattc tgtaatgatc ccttcctggg 1741 cgtctattat cacaagaaca acaaatcctg gatggagtcc gaatttagag tctactccag 1801 cgccaacaac tgcactttcg aatacgtatc acagccattc ttgatggacc ttgaaggaaa 1861 gcagggtaat ttcaagaact tgagggagtt cgtattcaag aatatcgacg ggtactttaa 1921 gatttatagc aaacacacac ccattaattt ggtgcgggat cttcctcagg gatttagtgc 1981 tcttgagcct ctcgttgacc tccctattgg cattaacatc acccgctttc aaaccctgtt 2041 ggccctgcat cggtcctacc tgacaccggg cgactcaagt tccggatgga ccgcaggtgc 2101 cgccgcatac tatgtgggct accttcagcc aagaacattt ctgctgaaat ataatgagaa 2161 cgggaccatt acagatgcgg tggattgtgc actcgaccct ctgtctgaga cgaaatgcac 2221 ccttaagagc ttcacggtgg agaaaggcat ttatcagact tctaacttca gagttcaacc 2281 caccgagtcc attgtgcgat tcccaaatat tacgaatttg tgcccatttg gtgaggtctt 2341 caatgctact cgattcgcct cagtttatgc atggaaccga aagagaattt ccaattgtgt 2401 ggcggactac tcagtattgt ataatagtgc aagctttagc acattcaaat gttacggcgt 2461 gtctccaacg aagctgaacg atctctgttt cacaaacgtt tatgcggatt ccttcgtgat 2521 tcgcggcgat gaggtccgac agattgcgcc tgggcaaacg ggtaagatcg ctgattacaa 2581 ctataagttg ccggacgatt tcacaggatg tgtcatagct tggaatagca ataatttgga 2641 cagtaaggtt ggcggaaact acaattattt gtacaggttg tttcgcaagt caaatttgaa 2701 accatttgag agagatatat ctacggagat atatcaagcc ggctctacac catgtaatgg 2761 tgtggagggc tttaactgct actttccact ccagtcatat ggtttccaac ctacaaatgg 2821 agtagggtat caaccgtaca gagttgtggt cttgagtttc gaattgctcc acgctccagc 2881 aacggtatgc ggtcctaaga aatccacaaa tcttgtgaag aacaagtgcg taaatttcaa 2941 cttcaatggg ctgactggaa caggcgtgct gactgagagt aacaagaagt tcttgccttt 3001 ccaacaattc gggcgggaca tagctgatac cactgacgcc gtccgcgacc ctcagaccct 3061 ggagattctg gacataactc cttgttcttt cggtggcgtc agtgttatca ctcccgggac 3121 caacacctcc aaccaagtcg cggtcctcta tcaaggcgtc aactgtacgg aagtaccggt 3181 agccatccat gcggaccaac ttacaccgac ttggagggtt tactctacag gaagcaatgt 3241 ctttcaaaca cgagccgggt gtctgatcgg agcagaacac gttaacaaca gctacgaatg 3301 tgacatacca ataggcgcag ggatttgtgc ttcatatcag acacagacca atagcccgcg 3361 gagagcgaga agcgtagcaa gccaaagcat catcgcgtac acgatgagcc tcggagcaga 3421 gaacagcgtc gcgtatagca ataattcaat agctatccca acaaatttca ctatttcggt 3481 caccactgag attctgccgg tctccatgac caagacatcc gtcgattgta ctatgtacat 3541 atgcggcgac agcacggagt gcagtaactt gctccttcag tacggttcct tctgtacgca 3601 gcttaaccgg gcactgacgg gtatcgcggt agaacaggac aagaacacac aggaggtctt 3661 cgcgcaggtc aaacaaatct acaagacacc acccataaag gacttcggcg ggttcaattt 3721 cagccaaatc ctgccggacc cttccaaacc tagtaagagg tcattcattg aggatcttct 3781 gtttaacaaa gttacgcttg cggacgcggg attcattaag cagtatggtg actgccttgg 3841 agatattgcc gccagggatt tgatatgtgc acagaaattt aacggcctca ccgttctgcc 3901 gcctctgctc accgatgaga tgatagcgca gtacacgagc gcactcctgg caggtacaat 3961 tacaagcgga tggacattcg gtgcaggagc agcgttgcag ataccctttg ctatgcagat 4021 ggcttatcga tttaacggga ttggcgtcac gcagaacgtc ctttatgaga atcagaaatt 4081 gattgcaaat cagttcaata gtgctatcgg taagattcag gacagcttga gcagtaccgc 4141 gtctgcactg ggaaagttgc aggacgtggt gaatcagaat gcacaagcac tgaatacctt 4201 ggttaagcaa ttgagtagca atttcggcgc catatcaagt gtactgaatg atatcctgtc 4261 acggttggac aaggtagaag ccgaagttca gattgaccgc ttgatcaccg ggcgcctcca 4321 aagtctgcag acctacgtca cacaacaatt gatcagagca gcagagataa gagcatctgc 4381 taacctggct gccactaaga tgtctgaatg tgtgcttggg cagtcaaaga gggtagattt 4441 ctgcggaaag ggctaccacc ttatgtcttt ccctcagagc gctccgcatg gtgtggtctt 4501 tctccatgtg acttatgtgc ctgctcaaga gaagaacttt acgacggcgc ccgctatatg 4561 ccatgatggt aaggcgcact ttccaaggga gggagtgttc gtgtccaacg gcactcactg 4621 gtttgtcacc caacgaaatt tctacgagcc tcaaattatt accaccgaca atacctttgt 4681 tagcggtaac tgtgacgtcg taattgggat tgttaataat acagtctacg atcctctgca 4741 gccggaactg gactccttta aagaggagct ggacaaatat ttcaagaacc acacatctcc 4801 tgacgtagat cttggagaca taagcggtat aaatgcaagt gttgttaaca ttcagaaaga 4861 aatagatagg ttgaacgaag ttgcgaagaa ccttaacgag tcactgatag acctccaaga 4921 gcttgggaag tacgagcaat atatcaagtg gccttggtat atttggctcg ggttcatagc 4981 aggacttatc gctatagtca tggtgactat aatgctgtgc tgcatgacaa gctgctgcag 5041 ctgtctcaaa ggctgttgct cttgcggctc ttaatgaaag cttatcgata ccgtcgacct 5101 cgagggccca gatctaattc accccaccag tgcaggctgc ctatcagaaa gtggtggctg 5161 gtgtggctaa tgccctggcc cacaagtatc actaagctcg ctttcttgct gtccaatttc 5221 tattaaaggt tcctttgttc cctaagtcca actactaaac tgggggatat tatgaagggc 5281 cttgagcatc tggattctgc ctaataaaaa acatttattt tcattgcaat gatgtattta 5341 aattatttct gaatatttta ctaaaaaggg aatgtgggag gtcagtgcat ttaaaacata 5401 aagaaatgaa gagctagttc aaaccttggg aaaatacact atatcttaaa ctccatgaaa 5461 gaaggtgagg ctgcaaacag ctaatgcaca ttggcaacag cccctgatgc ctatgcctta 5521 ttcatccctc agaaaaggat tcaagtagag gcttgatttg gaggttaaag ttttgctatg 5581 ctgtatttta cattacttat tgttttagct gtcctcatga atgtcttttc actacccatt 5641 tgcttatcct gcatctctca gccttgactc cactcagttc tcttgcttag agataccacc 5701 tttcccctga agtgttcctt ccatgtttta cggcgagatg gtttctcctc gcctggccac 5761 tcagccttag ttgtctctgt tgtcttatag aggtctactt gaagaaggaa aaacaggggg 5821 catggtttga ctgtcctgtg agcccttctt ccctgcctcc cccactcaca gtgacccgga 5881 atccctcgac atggcagtct agatcattct tgaagacgaa agggcctcgt gatacgccta 5941 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg 6001 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg 6061 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt 6121 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt 6181 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg 6241 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa 6301 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt 6361 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag 6421 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt 6481 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga 6541 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt 6601 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta 6661 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg 6721 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc 6781 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt 6841 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg 6901 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg 6961 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa 7021 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa 7081 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 7141 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 7201 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact 7261 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac 7321 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 7381 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 7441 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 7501 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc 7561 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 7621 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 7681 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc 7741 agcaacggag atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg 7801 ccaagggttg gtttgcgcat tcacagttct ccgcaagaat tgattggctc caattcttgg 7861 agtggtgaat ccgttagcga ggtgccgccg gcttccattc aggtcgaggt ggcccggctc 7921 catgcaccgc gacgcaacgc ggggaggcag acaaggtata gggcggcgcc tacaatccat 7981 gccaacccgt tccatgtgct cgccgaggcg gcataaatcg ccgtgacgat cagcggtcca 8041 atgatcgaag ttaggctggt aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg 8101 tcatctacct gcctggacag catggcctgc aacgcgggca tcccgatgcc gccggaagcg 8161 agaagaatca taatggggaa ggccatccag cctcgcgtcg gggagctttt tgcaaaagcc 8221 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc 8281 ggcctctgca taaataaaaa aaattagtca gccatg //