LOCUS Exported 8316 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Encodes SARS-CoV-2 Spike with 21 aa C-terminal deletions for lentiviral pseudotyping. ACCESSION . VERSION . KEYWORDS HDM-SARS2-Spike-delta21 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" primer_bind complement(144..163) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" enhancer 265..644 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 645..848 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 799..819 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 845..869 /label=LNCX /note="Human CMV promoter, forward primer" intron 962..1437 /label=-beta-globin intron /note="internally truncated intron from human beta-globin" primer_bind complement(5269..5285) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5270..5286) /label=pBluescriptKS /note="For pBluescript vector" polyA_signal 5379..5773 /label=-beta-globin poly(A) /note="human beta-globin polyadenylation signal" primer_bind complement(6107..6125) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6193..6297 /gene="bla" /label=AmpR promoter CDS 6298..7158 /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(6516..6535) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7329..7917 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7818..7837 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct 61 cgcgtcgggg agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg 121 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc 181 atgagcttgg cccattgcat acgttgtatc catatcataa tatgtacatt tatattggct 241 catgtccaac attaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa 301 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa 361 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 421 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 481 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 541 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 601 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 661 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 721 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 781 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 841 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 901 tccatagaag acaccgggac cgatccagcc tcccctcgaa gctgatcctg agaacttcag 961 ggtgagtcta tgggaccctt gatgttttct ttccccttct tttctatggt taagttcatg 1021 tcataggaag gggagaagta acagggtaca catattgacc aaatcagggt aattttgcat 1081 ttgtaatttt aaaaaatgct ttcttctttt aatatacttt tttgtttatc ttatttctaa 1141 tactttccct aatctctttc tttcagggca ataatgatac aatgtatcat gcctctttgc 1201 accattctaa agaataacag tgataatttc tgggttaagg caatagcaat atttctgcat 1261 ataaatattt ctgcatataa attgtaactg atgtaagagg tttcatattg ctaatagcag 1321 ctacaatcca gctaccattc tgcttttatt ttatggttgg gataaggctg gattattctg 1381 agtccaagct aggccctttt gctaatcatg ttcatacctc ttatcttcct cccacagctc 1441 ctgggcaacg tgctggtctg tgtgctggcc catcactttg gcaaagaatt ccgccaccat 1501 gttcgtgttc ctggtactcc ttcctttggt gtcttcccag tgtgtaaatc ttaccactcg 1561 gacccagctt ccacccgcct acaccaacag tttcacacgc ggcgtctact atcctgacaa 1621 ggtgtttagg agttcagtct tgcactcaac tcaagacttg ttcctccctt tctttagcaa 1681 tgtgacgtgg tttcatgcca ttcatgtctc cggcacaaac ggaacgaagc gctttgataa 1741 tcctgtgctc ccgttcaacg atggagtgta cttcgcgtcc acagagaaga gcaatatcat 1801 tcgaggttgg atcttcggaa cgacactcga ctcaaagacg cagtcccttc tcatcgtcaa 1861 taatgccacg aacgtggtca tcaaagtgtg cgagtttcaa ttctgtaatg atcccttcct 1921 gggcgtctat tatcacaaga acaacaaatc ctggatggag tccgaattta gagtctactc 1981 cagcgccaac aactgcactt tcgaatacgt atcacagcca ttcttgatgg accttgaagg 2041 aaagcagggt aatttcaaga acttgaggga gttcgtattc aagaatatcg acgggtactt 2101 taagatttat agcaaacaca cacccattaa tttggtgcgg gatcttcctc agggatttag 2161 tgctcttgag cctctcgttg acctccctat tggcattaac atcacccgct ttcaaaccct 2221 gttggccctg catcggtcct acctgacacc gggcgactca agttccggat ggaccgcagg 2281 tgccgccgca tactatgtgg gctaccttca gccaagaaca tttctgctga aatataatga 2341 gaacgggacc attacagatg cggtggattg tgcactcgac cctctgtctg agacgaaatg 2401 cacccttaag agcttcacgg tggagaaagg catttatcag acttctaact tcagagttca 2461 acccaccgag tccattgtgc gattcccaaa tattacgaat ttgtgcccat ttggtgaggt 2521 cttcaatgct actcgattcg cctcagttta tgcatggaac cgaaagagaa tttccaattg 2581 tgtggcggac tactcagtat tgtataatag tgcaagcttt agcacattca aatgttacgg 2641 cgtgtctcca acgaagctga acgatctctg tttcacaaac gtttatgcgg attccttcgt 2701 gattcgcggc gatgaggtcc gacagattgc gcctgggcaa acgggtaaga tcgctgatta 2761 caactataag ttgccggacg atttcacagg atgtgtcata gcttggaata gcaataattt 2821 ggacagtaag gttggcggaa actacaatta tttgtacagg ttgtttcgca agtcaaattt 2881 gaaaccattt gagagagata tatctacgga gatatatcaa gccggctcta caccatgtaa 2941 tggtgtggag ggctttaact gctactttcc actccagtca tatggtttcc aacctacaaa 3001 tggagtaggg tatcaaccgt acagagttgt ggtcttgagt ttcgaattgc tccacgctcc 3061 agcaacggta tgcggtccta agaaatccac aaatcttgtg aagaacaagt gcgtaaattt 3121 caacttcaat gggctgactg gaacaggcgt gctgactgag agtaacaaga agttcttgcc 3181 tttccaacaa ttcgggcggg acatagctga taccactgac gccgtccgcg accctcagac 3241 cctggagatt ctggacataa ctccttgttc tttcggtggc gtcagtgtta tcactcccgg 3301 gaccaacacc tccaaccaag tcgcggtcct ctatcaagac gtcaactgta cggaagtacc 3361 ggtagccatc catgcggacc aacttacacc gacttggagg gtttactcta caggaagcaa 3421 tgtctttcaa acacgagccg ggtgtctgat cggagcagaa cacgttaaca acagctacga 3481 atgtgacata ccaataggcg cagggatttg tgcttcatat cagacacaga ccaatagccc 3541 gcggagagcg agaagcgtag caagccaaag catcatcgcg tacacgatga gcctcggagc 3601 agagaacagc gtcgcgtata gcaataattc aatagctatc ccaacaaatt tcactatttc 3661 ggtcaccact gagattctgc cggtctccat gaccaagaca tccgtcgatt gtactatgta 3721 catatgcggc gacagcacgg agtgcagtaa cttgctcctt cagtacggtt ccttctgtac 3781 gcagcttaac cgggcactga cgggtatcgc ggtagaacag gacaagaaca cacaggaggt 3841 cttcgcgcag gtcaaacaaa tctacaagac accacccata aaggacttcg gcgggttcaa 3901 tttcagccaa atcctgccgg acccttccaa acctagtaag aggtcattca ttgaggatct 3961 tctgtttaac aaagttacgc ttgcggacgc gggattcatt aagcagtatg gtgactgcct 4021 tggagatatt gccgccaggg atttgatatg tgcacagaaa tttaacggcc tcaccgttct 4081 gccgcctctg ctcaccgatg agatgatagc gcagtacacg agcgcactcc tggcaggtac 4141 aattacaagc ggatggacat tcggtgcagg agcagcgttg cagataccct ttgctatgca 4201 gatggcttat cgatttaacg ggattggcgt cacgcagaac gtcctttatg agaatcagaa 4261 attgattgca aatcagttca atagtgctat cggtaagatt caggacagct tgagcagtac 4321 cgcgtctgca ctgggaaagt tgcaggacgt ggtgaatcag aatgcacaag cactgaatac 4381 cttggttaag caattgagta gcaatttcgg cgccatatca agtgtactga atgatatcct 4441 gtcacggttg gacaaggtag aagccgaagt tcagattgac cgcttgatca ccgggcgcct 4501 ccaaagtctg cagacctacg tcacacaaca attgatcaga gcagcagaga taagagcatc 4561 tgctaacctg gctgccacta agatgtctga atgtgtgctt gggcagtcaa agagggtaga 4621 tttctgcgga aagggctacc accttatgtc tttccctcag agcgctccgc atggtgtggt 4681 ctttctccat gtgacttatg tgcctgctca agagaagaac tttacgacgg cgcccgctat 4741 atgccatgat ggtaaggcgc actttccaag ggagggagtg ttcgtgtcca acggcactca 4801 ctggtttgtc acccaacgaa atttctacga gcctcaaatt attaccaccg acaatacctt 4861 tgttagcggt aactgtgacg tcgtaattgg gattgttaat aatacagtct acgatcctct 4921 gcagccggaa ctggactcct ttaaagagga gctggacaaa tatttcaaga accacacatc 4981 tcctgacgta gatcttggag acataagcgg tataaatgca agtgttgtta acattcagaa 5041 agaaatagat aggttgaacg aagttgcgaa gaaccttaac gagtcactga tagacctcca 5101 agagcttggg aagtacgagc aatatatcaa gtggccttgg tatatttggc tcgggttcat 5161 agcaggactt atcgctatag tcatggtgac tataatgctg tgctgcatga caagctgctg 5221 cagctgtctc aaaggctgtt gctcttgcgg ctcttaatga aagcttatcg ataccgtcga 5281 cctcgagggc ccagatctaa ttcaccccac cagtgcaggc tgcctatcag aaagtggtgg 5341 ctggtgtggc taatgccctg gcccacaagt atcactaagc tcgctttctt gctgtccaat 5401 ttctattaaa ggttcctttg ttccctaagt ccaactacta aactggggga tattatgaag 5461 ggccttgagc atctggattc tgcctaataa aaaacattta ttttcattgc aatgatgtat 5521 ttaaattatt tctgaatatt ttactaaaaa gggaatgtgg gaggtcagtg catttaaaac 5581 ataaagaaat gaagagctag ttcaaacctt gggaaaatac actatatctt aaactccatg 5641 aaagaaggtg aggctgcaaa cagctaatgc acattggcaa cagcccctga tgcctatgcc 5701 ttattcatcc ctcagaaaag gattcaagta gaggcttgat ttggaggtta aagttttgct 5761 atgctgtatt ttacattact tattgtttta gctgtcctca tgaatgtctt ttcactaccc 5821 atttgcttat cctgcatctc tcagccttga ctccactcag ttctcttgct tagagatacc 5881 acctttcccc tgaagtgttc cttccatgtt ttacggcgag atggtttctc ctcgcctggc 5941 cactcagcct tagttgtctc tgttgtctta tagaggtcta cttgaagaag gaaaaacagg 6001 gggcatggtt tgactgtcct gtgagccctt cttccctgcc tcccccactc acagtgaccc 6061 ggaatccctc gacatggcag tctagatcat tcttgaagac gaaagggcct cgtgatacgc 6121 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt 6181 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 6241 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg 6301 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt 6361 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga 6421 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa 6481 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt 6541 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt 6601 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc 6661 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga 6721 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat 6781 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct 6841 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc 6901 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg 6961 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc 7021 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg 7081 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca 7141 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta 7201 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 7261 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 7321 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 7381 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 7441 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 7501 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 7561 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 7621 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 7681 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 7741 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 7801 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 7861 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 7921 gccagcaacg gagatgcgcc gcgtgcggct gctggagatg gcggacgcga tggatatgtt 7981 ctgccaaggg ttggtttgcg cattcacagt tctccgcaag aattgattgg ctccaattct 8041 tggagtggtg aatccgttag cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg 8101 ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt atagggcggc gcctacaatc 8161 catgccaacc cgttccatgt gctcgccgag gcggcataaa tcgccgtgac gatcagcggt 8221 ccaatgatcg aagttaggct ggtaagagcc gcgagcgatc cttgaagctg tccctgatgg 8281 tcgtcatcta cctgcctgga cagcatggcc tgcaac //