LOCUS Exported 3714 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway-compatible Entry vector, with insert of NSP5 gene's CDS from SARS-CoV-2 isolate Wuhan-Hu-1. ACCESSION . VERSION . KEYWORDS pDONR223 SARS-CoV-2 NSP5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3714) 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 3714) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3714 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 79..106 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 198..284 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 333..355 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 347..364 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 348..364 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 380..479 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(1404..1503) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1520..1539) /label=T7 /note="T7 promoter, forward primer" promoter complement(1521..1539) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1544..1560) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1544..1560) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 1991..2782 /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 2875..3463 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3364..3383 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3617..3634 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga 61 agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc 121 ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc 181 aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata 241 aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc 301 tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc 361 cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga cctgttcgtt 421 gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa aagttggcat 481 gtccggattc cgcaagatgg ccttcccttc tggcaaggtg gagggctgca tggtgcaggt 541 gacctgcgga accaccaccc tgaacggtct gtggctggac gacgtggtgt actgccccag 601 gcacgtgatc tgcacctctg aggacatgct gaaccctaac tacgaagacc tgctgatcag 661 aaagagcaac cacaacttcc tggtgcaggc cggtaacgtg cagctgcgtg tgatcggcca 721 ctctatgcag aactgcgtgc tgaagctgaa ggtggacacc gctaacccca agacccctaa 781 gtacaagttc gtgagaatcc agcctggtca gaccttctcc gtgctggcct gctacaacgg 841 aagcccatcc ggcgtgtacc agtgcgctat gcgtcccaac ttcaccatca agggcagctt 901 cctgaacggc tcttgcggaa gcgtgggttt caacatcgac tacgactgcg tgtccttctg 961 ctacatgcac cacatggagc tgccaactgg agtgcacgct ggaactgacc tggagggtaa 1021 cttctacggc cccttcgtgg acagacagac tgctcaggct gctggcaccg acaccaccat 1081 caccgtgaac gtgctggctt ggctgtacgc cgctgtgatc aacggagacc gctggttcct 1141 gaacaggttc accaccaccc tgaacgactt caacctggtg gctatgaagt acaactacga 1201 gcctctgacc caggaccacg tggatatcct gggtccactg agcgcccaga ctggaatcgc 1261 tgtgctggac atgtgcgcct ccctgaagga actgctgcag aacggcatga acggacgcac 1321 catcctggga tccgctctgc tggaggacga attcaccccc ttcgacgtgg tgaggcagtg 1381 ctctggtgtg accttccagt aatacccaac tttcttgtac aaagttggca ttataagaaa 1441 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat 1501 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg 1561 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc 1621 atcatgcctc ctctagacca gccaggacag aaatgcctcg acttcgctgc tacccaaggt 1681 tgccgggtga cgcacaccgt ggaaacggat gaaggcacga acccagtgga cataagcctg 1741 ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca cgcaactggt ccagaacctt 1801 gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt 1861 ttttggggta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg 1921 atgtttgatg ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa 1981 gttaaacatt atgagggaag cggtgatcgc cgaagtatcg actcaactat cagaggtagt 2041 tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt acggctccgc 2101 agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg tgaccgtaag 2161 gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt cggcttcccc 2221 tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg acgacatcat 2281 tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc gcaatgacat 2341 tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct tgctgacaaa 2401 agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct ttgatccggt 2461 tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga actcgccgcc 2521 cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc 2581 agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc 2641 ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac aagaagaaga 2701 tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga aaggcgagat 2761 caccaaggta gtcggcaaat aaccctcgag ccacccatga ccaaaatccc ttaacgtgag 2821 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga 2881 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt 2941 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag 3001 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa 3061 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag 3121 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca 3181 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac 3241 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa 3301 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc 3361 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg 3421 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc 3481 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc 3541 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag 3601 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa 3661 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtt //