LOCUS Exported 8963 bp ds-DNA circular SYN 15-JUN-2021 DEFINITION Expresses TMPRSS2 in mammalian cells. ACCESSION . VERSION . KEYWORDS pscALPSblasti-TMPRSS2 Blasti SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8963) 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 8963) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 15, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8963 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 162..279 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 416..823 /label=SFFV promoter /note="spleen focus-forming virus long terminal repeat (LTR) promoter" regulatory 832..841 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 2811..2820 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3001..3012 /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /translation="WELQ" misc_feature 3225..3813 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3278..3298) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3696..3707) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 4331..4511 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(4582..5170) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4662..4681) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5341..6201) /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 5964..5983 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6202..6306) /gene="bla" /label=AmpR promoter primer_bind complement(6381..6400) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(6574..6594) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 6579..6908 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6759..6894 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6821..6840 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" enhancer 6920..7299 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 7301..7499 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 7454..7474 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 7517..7697 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 7744..7869 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 8366..8599 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 8783..8827 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" ORIGIN 1 tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag 61 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tcggcactgc 121 gtgcgccaat tctgcagaca aatggcagta ttcatccaca attttaaaag aaaagggggg 181 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact 241 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc 301 agagatccag tttggttaat taactgcagc cccgataaaa taaaagattt tatttagtct 361 ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag ctgcagtaac 421 gccattttgc aaggcatgga aaaataccaa accaagaata gagaagttca gatcaagggc 481 gggtacatga aaatagctaa cgttgggcca aacaggatat ctgcggtgag cagtttcggc 541 cccggcccgg ggccaagaac agatggtcac cgcagtttcg gccccggccc gaggccaaga 601 acagatggtc cccagatatg gcccaaccct cagcagtttc ttaagaccca tcagatgttt 661 ccaggctccc ccaaggacct gaaatgaccc tgcgccttat ttgaattaac caatcagcct 721 gcttctcgct tctgttcgcg cgcttctgct tcccgagctc tataaaagag ctcacaaccc 781 ctcactcggc gcgccagtcc tccgacagac tgagtcgccc gggggtctag agccaccatg 841 gctttgaact cagggtcacc accagctatt ggaccttact atgaaaacca tggataccaa 901 ccggaaaacc cctatcccgc acagcccact gtggtcccca ctgtctacga ggtgcatccg 961 gctcagtact acccgtcccc cgtgccccag tacgccccga gggtcctgac gcaggcttcc 1021 aaccccgtcg tctgcacgca gcccaaatcc ccatccggga cagtgtgcac ctcaaagact 1081 aagaaagcac tgtgcatcac cttgaccctg gggaccttcc tcgtgggagc tgcgctggcc 1141 gctggcctac tctggaagtt catgggcagc aagtgctcca actctgggat agagtgcgac 1201 tcctcaggta cctgcatcaa cccctctaac tggtgtgatg gcgtgtcaca ctgccccggc 1261 ggggaggacg agaatcggtg tgttcgcctc tacggaccaa acttcatcct tcaggtgtac 1321 tcatctcaga ggaagtcctg gcaccctgtg tgccaagacg actggaacga gaactacggg 1381 cgggcggcct gcagggacat gggctataag aataattttt actctagcca aggaatagtg 1441 gatgacagcg gatccaccag ctttatgaaa ctgaacacaa gtgccggcaa tgtcgatatc 1501 tataaaaaac tgtaccacag tgatgcctgt tcttcaaaag cagtggtttc tttacgctgt 1561 atagcctgcg gggtcaactt gaactcaagc cgccagagca ggatcgtggg cggcgagagc 1621 gcgctcccgg gggcctggcc ctggcaggtc agcctgcacg tccagaacgt ccacgtgtgc 1681 ggaggctcca tcatcacccc cgagtggatc gtgacagccg cccactgcgt ggaaaaacct 1741 cttaacaatc catggcattg gacggcattt gcggggattt tgagacaatc tttcatgttc 1801 tatggagccg gataccaagt agaaaaagtg atttctcatc caaattatga ctccaagacc 1861 aagaacaatg acattgcgct gatgaagctg cagaagcctc tgactttcaa cgacctagtg 1921 aaaccagtgt gtctgcccaa cccaggcatg atgctgcagc cagaacagct ctgctggatt 1981 tccgggtggg gggccaccga ggagaaaggg aagacctcag aagtgctgaa cgctgccaag 2041 gtgcttctca ttgagacaca gagatgcaac agcagatatg tctatgacaa cctgatcaca 2101 ccagccatga tctgtgccgg cttcctgcag gggaacgtcg attcttgcca gggtgacagt 2161 ggagggcctc tggtcacttc gaagaacaat atctggtggc tgatagggga tacaagctgg 2221 ggttctggct gtgccaaagc ttacagacca ggagtgtacg ggaatgtgat ggtattcacg 2281 gactggattt atcgacaaat gagggcagac ggctaggaat tctcgacctt gactggcggc 2341 gcgaccttga ggcctgcgtt cgcctcagtt gccccctctg tgcaatgggg agacgcgcct 2401 catcgcttga caacggccga agagccgccg cgcttccgtc tcccgcgtgc gcgcgccatg 2461 ctgcccaccc ccgttccgca ctgaccctcc cccgtgcccc gcgtcccgta ctgccgcccc 2521 gccccgagtc ccatgccgca gccaccgcga cggagcccgc aggcgggaac ctgcctccgc 2581 gcgttagcgc gcacgcgcgc ctcatgtgtc gtccccatca gcgccggctt ccgtctatag 2641 gccagatgca ctgtcactct ggcgaagtcg cagacccgat tggccgggac ggaggcgcga 2701 gaccgggttg cgggcggggc cgaacgtggt ataaaagggg cgggaggcca ggctcgtgcc 2761 gttttgcaga cgccaccgcc gaggaaaacc gtgtactatt agccacgcgt gccgccatgg 2821 ctaaaccact gtcccaggaa gaatccacac tgatcgaacg agcaaccgct actatcaact 2881 caatcccaat ctccgaagac tactctgtgg cctccgccgc cctgagcagc gacggacgaa 2941 tcttcaccgg cgtgaacgtc taccacttta ccggcggccc ctgcgctgag ctggtggtcc 3001 tgggaactgc agccgctgca gccgctggaa atctgacctg tatcgtggct atcgggaacg 3061 aaaatagggg aatcctgtct ccatgcggcc ggtgtagaca ggtcctgctg gatctgcatc 3121 ccgggatcaa ggcaattgtg aaagactcag atggacagcc cacagccgtc ggcattcgcg 3181 agctgctgcc tagcggctat gtgtgggaag ggtgagcgat cgctaatcaa cctctggatt 3241 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg 3301 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct 3361 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc 3421 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca 3481 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac 3541 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt 3601 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct 3661 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc 3721 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga 3781 cgagtcggat ctccctttgg gccgcctccc cgcttaatcg cgtcgagacc tagaaaaaca 3841 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc 3901 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac 3961 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 4021 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta 4081 cttccctgat tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg 4141 atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga 4201 gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt 4261 attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc 4321 ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact 4381 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc 4441 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa 4501 aatctctagc agggcccgtt tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 4561 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 4621 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 4681 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 4741 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 4801 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 4861 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 4921 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 4981 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 5041 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 5101 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 5161 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 5221 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 5281 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 5341 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 5401 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 5461 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 5521 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 5581 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 5641 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 5701 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 5761 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 5821 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 5881 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 5941 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 6001 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 6061 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 6121 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 6181 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 6241 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 6301 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat 6361 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg 6421 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca 6481 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg 6541 cttcgcgatg tacgggccag atatacgcgc tgtggaatgt gtgtcagtta gggtgtggaa 6601 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 6661 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 6721 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 6781 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 6841 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 6901 tttgcaaaaa gctcgcgttg acattgatta ttgactagtt attaatagta atcaattacg 6961 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc 7021 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc 7081 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact 7141 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat 7201 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact 7261 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac 7321 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac 7381 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac 7441 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcagc 7501 gcgttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg 7561 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt 7621 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt 7681 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag 7741 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg 7801 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc 7861 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg 7921 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa 7981 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga 8041 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta 8101 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac 8161 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cggccgctga 8221 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata 8281 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc 8341 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag 8401 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg 8461 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc 8521 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc 8581 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg 8641 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga 8701 cctggatgga gtgggacaga gaattaacaa ttacacaagc ttaatacact ccttaattga 8761 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc 8821 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat 8881 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag 8941 agttaggcag ggatattcac cat //