LOCUS Exported 3480 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway-compatible Entry vector. ACCESSION . VERSION . KEYWORDS pDONR223 SARS-CoV2 TEV-M SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3480) 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 3480) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3480 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 118..137 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 371..388 /label=L4440 /note="L4440 vector, forward primer" terminator 547..574 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 666..752 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 801..823 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 815..832 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 816..832 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 848..947 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 947..967 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" protein_bind complement(1638..1737) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1754..1773) /label=T7 /note="T7 promoter, forward primer" promoter complement(1755..1773) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1778..1794) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1778..1794) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 2225..3016 /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 3109..217 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 61 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 241 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga 301 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac 361 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc 421 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa 481 agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga 541 aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat 601 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc 661 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc 721 cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt 781 aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct 841 cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga 901 tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gttggcgaaa acctgtactt 961 ccaaggcatg gctgactcta acggtaccat caccgtggag gaactgaaga agctgctgga 1021 gcagtggaac ctggtcatcg gcttcctgtt cctgacctgg atctgcctgc tgcagttcgc 1081 ctacgctaac cgcaacaggt tcctgtacat catcaagctg atcttcctgt ggctgctgtg 1141 gcctgtgacc ctggcttgct tcgtgctggc tgccgtgtac cgcatcaact ggatcaccgg 1201 cggaatcgcc atcgctatgg cctgcctggt gggcctgatg tggctgtctt acttcatcgc 1261 tagcttcagg ctgttcgcca gaacccgttc catgtggtct ttcaaccccg agaccaacat 1321 cctgctgaac gtgcctctgc acggaaccat cctgaccaga ccactgctgg agagcgaact 1381 ggtcatcggc gctgtgatcc tgagaggaca cctgcgtatc gccggacacc acctgggtcg 1441 ttgcgacatc aaggacctgc ccaaggaaat caccgtggct accagccgca ccctgtccta 1501 ctacaagctg ggagcttctc agagagtggc tggtgactct ggtttcgctg cttactctcg 1561 ctacaggatc ggtaactaca agctgaacac cgaccacagc tcctctagcg acaacatcgc 1621 cctgctggtg cagtaatacc caactttctt gtacaaagtt ggcattataa gaaagcattg 1681 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca 1741 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct 1801 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg 1861 cctcctctag accagccagg acagaaatgc ctcgacttcg ctgctaccca aggttgccgg 1921 gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag tggacataag cctgttcggt 1981 tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga 2041 acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg 2101 ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt 2161 gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa 2221 cattatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt 2281 catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga 2341 tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga 2401 tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga 2461 gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg 2521 gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc 2581 aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag 2641 agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga 2701 acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg 2761 ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac 2821 cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca 2881 gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt 2941 ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa 3001 ggtagtcggc aaataaccct cgagccaccc atgaccaaaa tcccttaacg tgagttacgc 3061 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 3121 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 3181 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 3241 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt 3301 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 3361 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 3421 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact //