LOCUS Exported 9206 bp ds-DNA circular SYN 08-JUL-2021 DEFINITION expressing SARS-CoV-2 B.1.617.1 spike protein for pseudovirus production. ACCESSION . VERSION . KEYWORDS pcDNA3.3-SARS2-B.1.617.1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9206) TITLE SARS-CoV-2 (variant) pseudovirus plasmid JOURNAL Unpublished REFERENCE 2 (bases 1 to 9206) AUTHORS . TITLE Direct Submission JOURNAL Exported Jul 8, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9206 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 8..387 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 388..591 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 542..562 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 588..612 /label=LNCX /note="Human CMV promoter, forward primer" CDS complement(2030..2047) /codon_start=1 /product="tetracysteine peptide that binds biarsenical labeling reagents" /label=tetracysteine tag /translation="CCPGCC" primer_bind complement(4550..4569) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4594..4642 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4844..5272 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4931..4950) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5141..5162 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5281..5301) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5286..5615 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5466..5601 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5528..5547 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5682..6476 /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(5736..5755) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6346..6365 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6652..6773 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6689..6708) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6743..6762 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6822..6838) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6822..6838) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6835..6857) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6846..6862 /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(6870..6900) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6915..6936 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7053..7070) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7224..7812) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7304..7323) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7983..8843) /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 8606..8625 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8844..8948) /gene="bla" /label=AmpR promoter primer_bind complement(9023..9042) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt 61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga 121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca 181 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca 241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg 301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc 361 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt 421 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt 481 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg 541 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg 601 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg 661 gaccgatcca gcctccggac tgagctcctt aaggttaact ctagagaatt cggatccatg 721 tttgtgttcc tggtgctgct gccactggtg tccagccagt gtgtgaacct gaccaccagg 781 acccaacttc ctcctgccta caccaactcc ttcaccaggg gagtctacta ccctgacaag 841 gtgttcaggt cctctgtgct gcacagcacc caggacctgt tcctgccatt cttcagcaat 901 gtgacctggt tccatgccat ccatgtgtct ggcaccaatg gcaccaagag gtttgacaac 961 cctgtgctgc cattcaatga tggagtctac tttgccagca cagagaagag caacatcatc 1021 aggggctgga tttttggcac caccctggac agcaagaccc agtccctgct gattgtgaac 1081 aatgccacca atgtggtgat taaggtgtgt gagttccagt tctgtaatga cccattcctg 1141 gacgtctact accacaagaa caacaagtcc tggatgaagt ctgagttcag ggtctactcc 1201 tctgccaaca actgtacctt tgaatatgtg agccaaccat tcctgatgga cttggagggc 1261 aagcagggca acttcaagaa cctgagggag tttgtgttca agaacattga tggctacttc 1321 aagatttaca gcaaacacac accaatcaac ctggtgaggg acctgccaca gggcttctct 1381 gccttggaac cactggtgga cctgccaatt ggcatcaaca tcaccaggtt ccagaccctg 1441 ctggctctgc acaggtccta cctgacacct ggagactcct cctctggctg gacagcagga 1501 gcagcagcct actatgtggg ctacctccaa ccaaggacct tcctgctgaa atacaatgag 1561 aatggcacca tcacagatgc tgtggactgt gccctggacc cactgtctga gaccaagtgt 1621 accctgaaat ccttcacagt ggagaagggc atctaccaga ccagcaactt cagggtccaa 1681 ccaacagaga gcattgtgag gtttccaaac atcaccaacc tgtgtccatt tggagaggtg 1741 ttcaatgcca ccaggtttgc ctctgtctat gcctggaaca ggaagaggat tagcaactgt 1801 gtggctgact actctgtgct ctacaactct gcctccttca gcaccttcaa gtgttatgga 1861 gtgagcccaa ccaaactgaa tgacctgtgt ttcaccaatg tctatgctga ctcctttgtg 1921 attaggggag atgaggtgag acagattgcc cctggacaaa caggcaagat tgctgactac 1981 aactacaaac tgcctgatga cttcacaggc tgtgtgattg cctggaacag caacaacctg 2041 gacagcaagg tgggaggcaa ctacaactac aggtacagac tgttcaggaa gagcaacctg 2101 aaaccatttg agagggacat cagcacagag atttaccagg ctggcagcac accatgtaat 2161 ggagtgcagg gcttcaactg ttactttcca ctccaatcct atggcttcca accaaccaat 2221 ggagtgggct accaaccata cagggtggtg gtgctgtcct ttgaactgct ccatgcccct 2281 gccacagtgt gtggaccaaa gaagagcacc aacctggtga agaacaagtg tgtgaacttc 2341 aacttcaatg gactgacagg cacaggagtg ctgacagaga gcaacaagaa gttcctgcca 2401 ttccaacagt ttggcaggga cattgctgac accacagatg ctgtgaggga cccacagacc 2461 ttggagattc tggacatcac accatgttcc tttggaggag tgtctgtgat tacacctggc 2521 accaacacca gcaaccaggt ggctgtgctc taccagggtg tgaactgtac tgaggtgcct 2581 gtggctatcc atgctgacca acttacacca acctggaggg tctacagcac aggcagcaat 2641 gtgttccaga ccagggctgg ctgtctgatt ggagcagagc atgtgaacaa ctcctatgag 2701 tgtgacatcc caattggagc aggcatctgt gcctcctacc agacccagac caacagcaga 2761 aggagggcaa ggtctgtggc aagccagagc atcattgcct acacaatgag tctgggagca 2821 gagaactctg tggcttacag caacaacagc attgccatcc caaccaactt caccatctct 2881 gtgaccacag agattctgcc tgtgagtatg accaagacct ctgtggactg tacaatgtat 2941 atctgtggag acagcacaga gtgtagcaac ctgctgctcc aatatggctc cttctgtacc 3001 caacttaaca gggctctgac aggcattgct gtggaacagg acaagaacac ccaggaggtg 3061 tttgcccagg tgaagcagat ttacaagaca cctccaatca aggactttgg aggcttcaac 3121 ttcagccaga ttctgcctga cccaagcaag ccaagcaaga ggtccttcat tgaggacctg 3181 ctgttcaaca aggtgaccct ggctgatgct ggcttcatca agcaatatgg agactgtctg 3241 ggagacattg ctgccaggga cctgatttgt gcccagaagt tcaatggact gacagtgctg 3301 cctccactgc tgacagatga gatgattgcc caatacacct ctgccctgct ggctggcacc 3361 atcacctctg gctggacctt tggagcagga gcagccctcc aaatcccatt tgctatgcag 3421 atggcttaca ggttcaatgg cattggagtg acccagaatg tgctctatga gaaccagaaa 3481 ctgattgcca accagttcaa ctctgccatt ggcaagattc aggactccct gtccagcaca 3541 gcctctgccc tgggcaaact ccaagatgtg gtgaaccaga atgcccaggc tctgaacacc 3601 ctggtgaagc aactttccag caactttgga gccatctcct ctgtgctgaa tgacatcctg 3661 agcagactgg acaaggtgga ggctgaggtc cagattgaca gactgattac aggcagactc 3721 caatccctcc aaacctatgt gacccaacaa cttatcaggg ctgctgagat tagggcatct 3781 gccaacctgg ctgccaccaa gatgagtgag tgtgtgctgg gacaaagcaa gagggtggac 3841 ttctgtggca agggctacca cctgatgagt tttccacagt ctgcccctca tggagtggtg 3901 ttcctgcatg tgacctatgt gcctgcccat gagaagaact tcaccacagc ccctgccatc 3961 tgccatgatg gcaaggctca ctttccaagg gagggagtgt ttgtgagcaa tggcaccgac 4021 tggtttgtga cccagaggaa cttctatgaa ccacagatta tcaccacaga caacaccttt 4081 gtgtctggca actgtgatgt ggtgattggc attgtgaaca acacagtcta tgacccactc 4141 caacctgaac tggactcctt caaggaggaa ctggacaaat acttcaagaa ccacaccagc 4201 cctgatgtgg acctgggaga catctctggc atcaatgcct ctgtggtgaa catccagaag 4261 gagattgaca gactgaatga ggtggctaag aacctgaatg agtccctgat tgacctccaa 4321 gaactgggca aatatgaaca atacatcaag tggccatggt acatctggct gggcttcatt 4381 gctggactga ttgccattgt gatggtgacc ataatgctgt gttgtatgac ctcctgttgt 4441 tcctgtctga aaggctgttg ttcctgtggc tcctgttgta agtgactcga gccatggaag 4501 cttgatatct aactgactga accggtggta ccaaacgggg gaggctaact gaaacacgga 4561 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc 4621 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc 4681 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc 4741 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct 4801 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc 4861 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 4921 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 4981 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc 5041 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 5101 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 5161 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt 5221 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt 5281 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 5341 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag 5401 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc 5461 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 5521 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt 5581 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca 5641 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat 5701 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac 5761 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc 5821 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc 5881 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag 5941 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc 6001 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg 6061 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc 6121 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc 6181 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga 6241 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca 6301 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 6361 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 6421 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg 6481 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc 6541 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc 6601 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt 6661 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 6721 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc 6781 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg 6841 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 6901 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 6961 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 7021 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 7081 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 7141 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 7201 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 7261 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 7321 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 7381 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 7441 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 7501 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 7561 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 7621 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 7681 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 7741 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 7801 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 7861 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 7921 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 7981 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 8041 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 8101 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 8161 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 8221 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 8281 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 8341 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 8401 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 8461 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 8521 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 8581 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 8641 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 8701 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 8761 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 8821 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 8881 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 8941 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg 9001 atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc 9061 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa 9121 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc 9181 tgcttcgcga tgtacgggcc agatat //