LOCUS Exported 3234 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway-compatible Entry vector. ACCESSION . VERSION . KEYWORDS pDONR223 SARS-CoV2 TEV-NSP10 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3234) AUTHORS Kim DK, Knapp JJ, Kuang D, Chawla A, Cassonnet P, Lee H, Sheykhkarimli D, Samavarchi-Tehrani P, Abdouni H, Rayhan A, Li R, Pogoutse O, Coyaud E, van der Werf S, Demeret C, Gingras AC, Taipale M, Raught B, Jacob Y, Roth FP TITLE A Comprehensive, Flexible Collection of SARS-CoV-2 Coding Regions. JOURNAL G3 (Bethesda). 2020 Aug 6. pii: g3.120.401554. doi: 10.1534/g3.120.401554. PUBMED 32763951 REFERENCE 2 (bases 1 to 3234) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3234 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 82..109 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 201..287 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 336..358 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 350..367 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 351..367 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 383..482 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 482..502 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" protein_bind complement(927..1026) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1043..1062) /label=T7 /note="T7 promoter, forward primer" promoter complement(1044..1062) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1067..1083) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1067..1083) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 1514..2305 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 2398..2986 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2887..2906 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3140..3157 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca 61 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga 121 tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg 181 ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag 241 ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt 301 ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac 361 ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag tgacctgttc 421 gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca aaaaagttgg 481 cgaaaacctg tacttccaag gcatggctgg taacgctact gaggtgcctg ctaacagcac 541 cgtgctgtcc ttctgcgcct tcgctgtgga cgccgctaag gcctacaagg actacctggc 601 ttccggcgga cagcctatca ccaactgcgt gaagatgctg tgcacccaca ccggaaccgg 661 tcaggccatc accgtgaccc cagaggctaa catggaccag gaatctttcg gtggcgccag 721 ctgctgcctg tactgcagat gccacatcga ccaccccaac cctaagggtt tctgcgacct 781 gaagggcaag tacgtgcaga tcccaaccac ctgcgctaac gaccccgtgg gattcaccct 841 gaagaacacc gtgtgcaccg tgtgcggaat gtggaagggc tacggatgct cttgcgacca 901 gctgagggaa cctatgctgc agtaataccc aactttcttg tacaaagttg gcattataag 961 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat 1021 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc 1081 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat 1141 atcatcatgc ctcctctaga ccagccagga cagaaatgcc tcgacttcgc tgctacccaa 1201 ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt ggacataagc 1261 ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac 1321 cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg 1381 tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg 1441 tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac 1501 aaagttaaac attatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt 1561 agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc 1621 cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt 1681 aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc 1741 ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat 1801 cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga 1861 cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac 1921 aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc 1981 ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc 2041 gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag 2101 cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct 2161 gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga 2221 agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga 2281 gatcaccaag gtagtcggca aataaccctc gagccaccca tgaccaaaat cccttaacgt 2341 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg 2401 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc 2461 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag 2521 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa 2581 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc 2641 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc 2701 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta 2761 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag 2821 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct 2881 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga 2941 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc 3001 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt 3061 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg 3121 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg 3181 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acag //