LOCUS Exported 4596 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway-compatible Entry vector, with insert of NSP13 gene's CDS from SARS-CoV-2 isolate Wuhan-Hu-1. Does not contain stop codon. . ACCESSION . VERSION . KEYWORDS pDONR223 SARS-CoV-2 NSP13_nostop SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4596) 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 4596) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4596 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 129..156 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 248..334 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 383..405 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 397..414 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 398..414 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 430..529 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(2336..2435) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(2452..2471) /label=T7 /note="T7 promoter, forward primer" promoter complement(2453..2471) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2476..2492) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2476..2492) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 2923..3714 /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 3807..4395 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4296..4315 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4549..4566 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg 61 aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta 121 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt 181 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc 241 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg 301 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg 361 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag 421 ctcgggcccc aaataatgat tttattttga ctgatagtga cctgttcgtt gcaacaaatt 481 gatgagcaat gcttttttat aatgccaact ttgtacaaaa aagttggcat ggccgtgggc 541 gcttgcgtgc tgtgcaactc tcagaccagc ctgagatgtg gagcttgcat ccgcaggcct 601 ttcctgtgct gcaagtgctg ctacgaccac gtgatcagca cctcccacaa gctggtgctg 661 tccgtgaacc catacgtgtg caacgctcct ggttgtgacg tgactgacgt gacccagctg 721 tacctgggcg gaatgagcta ctactgcaag tcccacaagc cccctatcag cttcccactg 781 tgcgctaacg gccaggtgtt cggactgtac aagaacacct gcgtgggctc cgacaacgtg 841 accgacttca acgccatcgc tacctgcgac tggaccaacg ccggagacta catcctggct 901 aacacctgca ccgaaaggct gaagctgttc gccgctgaga ccctgaaggc caccgaggaa 961 accttcaagc tgtcttacgg tatcgctacc gtgcgtgagg tgctgtctga ccgcgaactg 1021 cacctgtctt gggaagtggg caagccaaga ccacccctga accgtaacta cgtgttcacc 1081 ggatacaggg tgaccaagaa cagcaaggtg cagatcggag agtacacctt cgaaaaggga 1141 gactacggtg acgccgtggt gtacagagga accaccacct acaagctgaa cgtgggtgac 1201 tacttcgtgc tgacctccca caccgtgatg ccactgtctg ctcccaccct ggtgcctcag 1261 gaacactacg tgcgcatcac cggactgtac ccaaccctga acatctccga cgagttcagc 1321 tccaacgtgg ccaactacca gaaagtgggc atgcagaagt actccaccct gcagggacct 1381 ccaggtaccg gcaagtctca cttcgccatc ggtctggctc tgtactaccc ctctgcccgt 1441 atcgtgtaca ctgcttgttc tcacgctgct gtggatgctc tgtgcgagaa ggctctgaag 1501 tacctgccta tcgacaagtg ctcccgcatc atcccagcca gagctcgtgt ggaatgcttc 1561 gacaagttca aggtgaactc taccctggag cagtacgtgt tctgcaccgt gaacgccctg 1621 cctgaaacca ccgctgacat cgtggtgttc gacgagatca gcatggccac caactacgac 1681 ctgtctgtgg tgaacgctag actgagggct aagcactacg tgtacatcgg tgacccagct 1741 cagctgcctg ctcctcgtac cctgctgacc aagggcaccc tggagccaga atacttcaac 1801 agcgtgtgcc gcctgatgaa gaccatcggt ccagacatgt tcctgggtac ctgccgccgc 1861 tgccctgctg aaatcgtgga caccgtgtct gctctggtgt acgacaacaa gctgaaggcc 1921 cacaaggaca agagcgccca gtgcttcaag atgttctaca agggagtgat cacccacgac 1981 gtgtctagcg ccatcaacag gccccagatc ggtgtggtgc gcgagttcct gaccaggaac 2041 cccgcctgga gaaaggccgt gttcatctcc ccttacaact ctcagaacgc cgtggcttcc 2101 aagatcctgg gtctgcctac ccagaccgtg gactcctctc agggctctga atacgactac 2161 gtgatcttca cccagaccac cgagaccgcc cacagctgca acgtgaacag gttcaacgtg 2221 gccatcacca gagctaaagt gggaatcctg tgcatcatga gcgacagaga cctgtacgac 2281 aagctgcagt tcacctccct ggagatcccc cgcaggaacg tggctaccct gcagtaccca 2341 actttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa 2401 caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt 2461 gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc 2521 tgatgttaca ttgcacaaga taaaaatata tcatcatgcc tcctctagac cagccaggac 2581 agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg 2641 atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag 2701 cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca 2761 gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca 2821 tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat 2881 gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc 2941 gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg 3001 acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt 3061 gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg 3121 atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta 3181 gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa 3241 ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg 3301 atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta 3361 ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta 3421 aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta 3481 gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat 3541 gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa 3601 gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg 3661 gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg 3721 agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac 3781 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 3841 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 3901 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta 3961 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 4021 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 4081 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 4141 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 4201 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 4261 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 4321 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 4381 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 4441 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc 4501 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg 4561 agcgaggaag cggaagagcg cccaatacgc aaaccg //