LOCUS Exported 3132 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway-compatible Entry vector, with insert of NSP9 gene's CDS from SARS-CoV-2 isolate Wuhan-Hu-1. Does not contain stop codon. . ACCESSION . VERSION . KEYWORDS pDONR223 SARS-CoV-2 NSP9_nostop SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3132) 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 3132) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3132 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 138..165 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 257..343 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 392..414 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 406..423 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 407..423 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 439..538 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(881..980) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(997..1016) /label=T7 /note="T7 promoter, forward primer" promoter complement(998..1016) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1021..1037) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1021..1037) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 1468..2259 /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 2352..2940 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2841..2860 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3094..3111 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt 61 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa 121 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc 181 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca 241 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa 301 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct 361 actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc 421 agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac ctgttcgttg 481 caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa agttggcatg 541 aacaacgagc tgagccctgt ggccctgaga cagatgtctt gtgctgctgg taccacccag 601 accgcttgca ccgacgacaa cgccctggct tactacaaca ccaccaaggg cggaagattc 661 gtgctggccc tgctgtctga cctgcaggac ctgaagtggg ctcgtttccc caagagcgac 721 ggtaccggca ccatctacac cgagctggaa cccccttgca gattcgtgac cgacacccca 781 aagggcccca aggtgaagta cctgtacttc atcaagggac tgaacaacct gaacagagga 841 atggtgctgg gtagcctggc tgctaccgtg aggctgcagt acccaacttt cttgtacaaa 901 gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt 961 caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg 1021 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca 1081 caagataaaa atatatcatc atgcctcctc tagaccagcc aggacagaaa tgcctcgact 1141 tcgctgctac ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc 1201 cagtggacat aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc 1261 aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg 1321 gcttgttatg actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg 1381 cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc 1441 agtcgcccta aaacaaagtt aaacattatg agggaagcgg tgatcgccga agtatcgact 1501 caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta 1561 catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg 1621 gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg 1681 gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt 1741 gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa 1801 tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg 1861 gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag 1921 gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg 1981 ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc 2041 cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg 2101 gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat 2161 cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac 2221 tacgtgaaag gcgagatcac caaggtagtc ggcaaataac cctcgagcca cccatgacca 2281 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc 2341 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 2401 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 2461 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta 2521 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 2581 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 2641 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 2701 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 2761 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 2821 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 2881 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 2941 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 3001 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct 3061 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa 3121 gagcgcccaa ta //