LOCUS Exported 8816 bp ds-DNA circular SYN 17-NOV-2021 DEFINITION Transient mammalian expression of Luc2 (firefly luciferase) along with SARS-CoV-2 sequence 20080-21171 (PS9) within the 3'UTR. Resulting transcript is packaged into SARS-CoV-2 virus-like particles.. ACCESSION . VERSION . KEYWORDS Luc-PS9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8816) AUTHORS Syed AM, Taha TY, Tabata T, Chen IP, Ciling A, Khalid MM, Sreekumar B, Chen PY, Hayashi JM, Soczek KM, Ott M, Doudna JA TITLE Rapid assessment of SARS-CoV-2 evolved variants using virus-like particles. JOURNAL Science. 2021 Nov 4:eabl6184. doi: 10.1126/science.abl6184. PUBMED 34735219 REFERENCE 2 (bases 1 to 8816) AUTHORS . TITLE Direct Submission JOURNAL Exported Nov 17, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8816 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 15..394 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 395..598 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 549..569 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 643..662 /label=T7 /note="T7 promoter, forward primer" promoter 643..661 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 681..690 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 687..2339 /codon_start=1 /product="firefly luciferase" /label=luciferase /note="synthetic luc2 version of the luciferase gene" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" misc_feature 3530..4118 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3583..3603) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4001..4012) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(4187..4204) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4193..4417 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4463..4891 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4550..4569) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4760..4781 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4900..4920) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4905..5234 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5085..5220 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5147..5166 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5301..6095 /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(5355..5374) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5965..5984 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6269..6390 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6306..6325) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6360..6379 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6439..6455) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6439..6455) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6452..6474) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6463..6479 /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(6487..6517) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6532..6553 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6670..6687) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6841..7429) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6921..6940) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7600..8460) /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 8223..8242 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8461..8565) /gene="bla" /label=AmpR promoter primer_bind complement(8640..8659) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc 61 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc 121 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt 181 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca 241 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg 301 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca 361 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa 421 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa 481 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc 541 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct 601 ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag 661 ggagacccaa gctggctagc gccgccatgg aagatgccaa aaacattaag aagggcccag 721 cgccattcta cccactcgaa gacgggaccg ccggcgagca gctgcacaaa gccatgaagc 781 gctacgccct ggtgcccggc accatcgcct ttaccgacgc acatatcgag gtggacatta 841 cctacgccga gtacttcgag atgagcgttc ggctggcaga agctatgaag cgctatgggc 901 tgaatacaaa ccatcggatc gtggtgtgca gcgagaatag cttgcagttc ttcatgcccg 961 tgttgggtgc cctgttcatc ggtgtggctg tggccccagc taacgacatc tacaacgagc 1021 gcgagctgct gaacagcatg ggcatcagcc agcccaccgt cgtattcgtg agcaagaaag 1081 ggctgcaaaa gatcctcaac gtgcaaaaga agctaccgat catacaaaag atcatcatca 1141 tggatagcaa gaccgactac cagggcttcc aaagcatgta caccttcgtg acttcccatt 1201 tgccacccgg cttcaacgag tacgacttcg tgcccgagag cttcgaccgg gacaaaacca 1261 tcgccctgat catgaacagt agtggcagta ccggattgcc caagggcgta gccctaccgc 1321 accgcaccgc ttgtgtccga ttcagtcatg cccgcgaccc catcttcggc aaccagatca 1381 tccccgacac cgctatcctc agcgtggtgc catttcacca cggcttcggc atgttcacca 1441 cgctgggcta cttgatctgc ggctttcggg tcgtgctcat gtaccgcttc gaggaggagc 1501 tattcttgcg cagcttgcaa gactataaga ttcaatctgc cctgctggtg cccacactat 1561 ttagcttctt cgctaagagc actctcatcg acaagtacga cctaagcaac ttgcacgaga 1621 tcgccagcgg cggggcgccg ctcagcaagg aggtaggtga ggccgtggcc aaacgcttcc 1681 acctaccagg catccgccag ggctacggcc tgacagaaac aaccagcgcc attctgatca 1741 cccccgaagg ggacgacaag cctggcgcag taggcaaggt ggtgcccttc ttcgaggcta 1801 aggtggtgga cttggacacc ggtaagacac tgggtgtgaa ccagcgcggc gagctgtgcg 1861 tccgtggccc catgatcatg agcggctacg ttaacaaccc cgaggctaca aacgctctca 1921 tcgacaagga cggctggctg cacagcggcg acatcgccta ctgggacgag gacgagcact 1981 tcttcatcgt ggaccggctg aagagcctga tcaaatacaa gggctaccag gtagccccag 2041 ccgaactgga gagcatcctg ctgcaacacc ccaacatctt cgacgccggg gtcgccggcc 2101 tgcccgacga cgatgccggc gagctgcccg ccgcagtcgt cgtgctggaa cacggtaaaa 2161 ccatgaccga gaaggagatc gtggactatg tggccagcca ggttacaacc gccaagaagc 2221 tgcgcggtgg tgttgtgttc gtggacgagg tgcctaaagg actgaccggc aagttggacg 2281 cccgcaagat ccgcgagatt ctcattaagg ccaagaaggg cggcaagatc gccgtgtaag 2341 aattcgctta tgcggccgcc tcctttaaga aggagatata gatctctgta ggtcccaaac 2401 aagctagtct taatggagtc acattaattg gagaagccgt aaaaacacag ttcaattatt 2461 ataagaaagt tgatggtgtt gtccaacaat tacctgaaac ttactttact cagagtagaa 2521 atttacaaga atttaaaccc aggagtcaaa tggaaattga tttcttagaa ttagctatgg 2581 atgaattcat tgaacggtat aaattagaag gctatgcctt cgaacatatc gtttatggag 2641 attttagtca tagtcagtta ggtggtttac atctactgat tggactagct aaacgtttta 2701 aggaatcacc ttttgaatta gaagatttta ttcctatgga cagtacagtt aaaaactatt 2761 tcataacaga tgcgcaaaca ggttcatcta agtgtgtgtg ttctgttatt gatttattac 2821 ttgatgattt tgttgaaata ataaaatccc aagatttatc tgtagtttct aaggttgtca 2881 aagtgactat tgactataca gaaatttcat ttatgctttg gtgtaaagat ggccatgtag 2941 aaacatttta cccaaaatta caatctagtc aagcgtggca accgggtgtt gctatgccta 3001 atctttacaa aatgcaaaga atgctattag aaaagtgtga ccttcaaaat tatggtgata 3061 gtgcaacatt acctaaaggc ataatgatga atgtcgcaaa atatactcaa ctgtgtcaat 3121 atttaaacac attaacatta gctgtaccct ataatatgag agttatacat tttggtgctg 3181 gttctgataa aggagttgca ccaggtacag ctgttttaag acagtggttg cctacgggta 3241 cgctgcttgt cgattcagat cttaatgact ttgtctctga tgcagattca actttgattg 3301 gtgattgtgc aactgtacat acagctaata aatgggatct cattattagt gatatgtacg 3361 accctaagac taaaaatgtt acaaaagaaa atgactctaa agagggtttt ttcacttaca 3421 tttgtgggtt tatacaacaa aagctagctc ttggaggttc cgtggctata aagatagatc 3481 ccaactccat aagtgtgtcg acatataagg atccactcga cgactcgaca atcaacctct 3541 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct 3601 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat 3661 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt 3721 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat 3781 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc 3841 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga 3901 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc 3961 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga 4021 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcatcttc gccttcgccc 4081 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattaatt cgagctcggt 4141 accttgattc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct 4201 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt 4261 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg 4321 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac 4381 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc 4441 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg 4501 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct 4561 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg 4621 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag 4681 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg 4741 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc 4801 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat 4861 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt 4921 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 4981 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 5041 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc 5101 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 5161 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 5221 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga 5281 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg 5341 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg 5401 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt 5461 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg 5521 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat 5581 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat 5641 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg 5701 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg 5761 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc 5821 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc 5881 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg 5941 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg 6001 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca 6061 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac 6121 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga 6181 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga 6241 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa 6301 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 6361 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccctcca cctctagcta 6421 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6481 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6541 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 6601 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 6661 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 6721 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 6781 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 6841 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 6901 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 6961 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 7021 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 7081 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 7141 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7201 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7261 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac 7321 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7381 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7441 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7501 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7561 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7621 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7681 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 7741 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 7801 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 7861 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 7921 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 7981 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 8041 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 8101 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 8161 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 8221 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8281 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8341 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8401 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8461 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8521 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8581 agtgccacct gacctccacg gatcgggaga tctcccgatc ccctatggtg cactctcagt 8641 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag 8701 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat 8761 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggc //