LOCUS Exported 3711 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. Does not contain stop codon. . ACCESSION . VERSION . KEYWORDS pDONR223 SARS-CoV-2 NSP5_nostop SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3711) 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 3711) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3711 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 118..145 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 237..323 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 372..394 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 386..403 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 387..403 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 419..518 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(1440..1539) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1556..1575) /label=T7 /note="T7 promoter, forward primer" promoter complement(1557..1575) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1580..1596) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1580..1596) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 2027..2818 /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 2911..3499 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3400..3419 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3653..3670 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag 61 tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa 121 aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt 181 cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt 241 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc 301 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc 361 atggatgttt tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggcccca 421 aataatgatt ttattttgac tgatagtgac ctgttcgttg caacaaattg atgagcaatg 481 cttttttata atgccaactt tgtacaaaaa agttggcatg tccggattcc gcaagatggc 541 cttcccttct ggcaaggtgg agggctgcat ggtgcaggtg acctgcggaa ccaccaccct 601 gaacggtctg tggctggacg acgtggtgta ctgccccagg cacgtgatct gcacctctga 661 ggacatgctg aaccctaact acgaagacct gctgatcaga aagagcaacc acaacttcct 721 ggtgcaggcc ggtaacgtgc agctgcgtgt gatcggccac tctatgcaga actgcgtgct 781 gaagctgaag gtggacaccg ctaaccccaa gacccctaag tacaagttcg tgagaatcca 841 gcctggtcag accttctccg tgctggcctg ctacaacgga agcccatccg gcgtgtacca 901 gtgcgctatg cgtcccaact tcaccatcaa gggcagcttc ctgaacggct cttgcggaag 961 cgtgggtttc aacatcgact acgactgcgt gtccttctgc tacatgcacc acatggagct 1021 gccaactgga gtgcacgctg gaactgacct ggagggtaac ttctacggcc ccttcgtgga 1081 cagacagact gctcaggctg ctggcaccga caccaccatc accgtgaacg tgctggcttg 1141 gctgtacgcc gctgtgatca acggagaccg ctggttcctg aacaggttca ccaccaccct 1201 gaacgacttc aacctggtgg ctatgaagta caactacgag cctctgaccc aggaccacgt 1261 ggatatcctg ggtccactga gcgcccagac tggaatcgct gtgctggaca tgtgcgcctc 1321 cctgaaggaa ctgctgcaga acggcatgaa cggacgcacc atcctgggat ccgctctgct 1381 ggaggacgaa ttcaccccct tcgacgtggt gaggcagtgc tctggtgtga ccttccagta 1441 cccaactttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa 1501 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta 1561 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa 1621 tctctgatgt tacattgcac aagataaaaa tatatcatca tgcctcctct agaccagcca 1681 ggacagaaat gcctcgactt cgctgctacc caaggttgcc gggtgacgca caccgtggaa 1741 acggatgaag gcacgaaccc agtggacata agcctgttcg gttcgtaagc tgtaatgcaa 1801 gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg gtggtaacgg 1861 cgcagtggcg gttttcatgg cttgttatga ctgttttttt ggggtacagt ctatgcctcg 1921 ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa 1981 cgatgttacg cagcagggca gtcgccctaa aacaaagtta aacattatga gggaagcggt 2041 gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc gccatctcga 2101 accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc tgaagccaca 2161 cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc 2221 tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga ttctccgcgc 2281 tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc cagctaagcg 2341 cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct tcgagccagc 2401 cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata gcgttgcctt 2461 ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc tatttgaggc 2521 gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg atgagcgaaa 2581 tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa tcgcgccgaa 2641 ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc ccgtcatact 2701 tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc gcgcagatca 2761 gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg gcaaataacc 2821 ctcgagccac ccatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc 2881 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg 2941 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct 3001 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct 3061 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct 3121 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg 3181 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc 3241 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga 3301 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 3361 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta 3421 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 3481 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 3541 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat 3601 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc 3661 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg c //