LOCUS Exported 3081 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway-compatible Entry vector, with insert of ORF9B gene's CDS from SARS-CoV-2 isolate Wuhan-Hu-1. Does not contain stop codon. . ACCESSION . VERSION . KEYWORDS pDONR223 SARS-CoV-2 ORF9B_nostop SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3081) 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 3081) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3081 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 55..141 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 190..212 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 204..221 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 205..221 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 237..336 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(628..727) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(744..763) /label=T7 /note="T7 promoter, forward primer" promoter complement(745..763) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(768..784) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(768..784) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 1215..2006 /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 2099..2687 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2588..2607 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2841..2858 /label=L4440 /note="L4440 vector, forward primer" terminator 3017..3044 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" ORIGIN 1 tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt 61 cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga 121 ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcgtta acgctagcat 181 ggatgttttc ccagtcacga cgttgtaaaa cgacggccag tcttaagctc gggccccaaa 241 taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat gagcaatgct 301 tttttataat gccaactttg tacaaaaaag ttggcatgga cccaaagatc tctgaaatgc 361 accccgctct gagactggtg gaccctcaga tccagctggc cgtgacccgc atggaaaacg 421 ctgtgggcag ggaccagaac aacgtgggac ccaaggtgta ccctatcatc ctgagactgg 481 gcagccctct gtccctgaac atggcccgta agaccctgaa cagcctggag gacaaggcct 541 tccagctgac cccaatcgct gtgcagatga ccaagctggc caccaccgag gaactgcccg 601 acgaattcgt ggtggtgacc gtgaagtacc caactttctt gtacaaagtt ggcattataa 661 gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca 721 ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt 781 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata 841 tatcatcatg cctcctctag accagccagg acagaaatgc ctcgacttcg ctgctaccca 901 aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag tggacataag 961 cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa 1021 ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact 1081 gtttttttgg ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg 1141 gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa 1201 caaagttaaa cattatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg 1261 tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct 1321 ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg 1381 taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt 1441 cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca 1501 tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg 1561 acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga 1621 caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc 1681 cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc 1741 cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca 1801 gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc 1861 tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag 1921 aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg 1981 agatcaccaa ggtagtcggc aaataaccct cgagccaccc atgaccaaaa tcccttaacg 2041 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt 2101 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag 2161 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca 2221 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca 2281 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg 2341 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg 2401 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct 2461 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga 2521 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 2581 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg 2641 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 2701 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt 2761 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc 2821 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac 2881 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc 2941 ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag 3001 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct 3061 ggcagtttat ggcgggcgtc c //