LOCUS Exported 4377 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway-compatible Entry vector, with insert of NSP14 gene's CDS from SARS-CoV-2 isolate Wuhan-Hu-1. ACCESSION . VERSION . KEYWORDS pDONR223 SARS-CoV-2 NSP14 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4377) 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 4377) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4377 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 83..110 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 202..288 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 337..359 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 351..368 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 352..368 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 384..483 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(2071..2170) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(2187..2206) /label=T7 /note="T7 promoter, forward primer" promoter complement(2188..2206) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2211..2227) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2211..2227) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 2658..3449 /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 3542..4130 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4031..4050 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4284..4301 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc 61 aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg 121 atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac 181 gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca 241 gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt 301 tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt tgtaaaacga 361 cggccagtct taagctcggg ccccaaataa tgattttatt ttgactgata gtgacctgtt 421 cgttgcaaca aattgatgag caatgctttt ttataatgcc aactttgtac aaaaaagttg 481 gcatggctga gaacgtgacc ggtctgttca aggactgctc taaggtcatc actggtctgc 541 acccaactca ggctccaacc cacctgtctg tggacaccaa gttcaagacc gaaggactgt 601 gcgtggacat ccccggtatc cctaaggaca tgacctaccg caggctgatc tccatgatgg 661 gcttcaagat gaactaccag gtgaacggat accctaacat gttcatcacc agagaggaag 721 ctatccgcca cgtgagggcc tggatcggtt tcgacgtgga aggttgccac gctaccagag 781 aagctgtggg taccaacctg ccactgcagc tgggcttctc taccggagtg aacctggtgg 841 ctgtgcctac cggctacgtg gacaccccaa acaacaccga cttctctaga gtgagcgcca 901 agccccctcc aggagaccag ttcaagcacc tgatccccct gatgtacaag ggtctgcctt 961 ggaacgtggt gcgcatcaag atcgtgcaga tgctgtccga caccctgaag aacctgtctg 1021 acagggtggt gttcgtgctg tgggctcacg gcttcgagct gaccagcatg aagtacttcg 1081 tgaagatcgg acccgaacgt acctgctgcc tgtgcgacag acgtgccacc tgcttctcca 1141 ccgcctctga cacctacgct tgctggcacc actccatcgg attcgactac gtgtacaacc 1201 ctttcatgat cgacgtgcag cagtggggat tcaccggtaa cctgcagtcc aaccacgacc 1261 tgtactgcca ggtgcacggt aacgcccacg tggcttcttg cgacgctatc atgaccagat 1321 gcctggccgt gcacgagtgc ttcgtgaagc gtgtggactg gaccatcgag taccctatca 1381 tcggcgacga actgaagatc aacgccgctt gcagaaaggt gcagcacatg gtggtgaagg 1441 ccgctctgct ggctgacaag ttcccagtgc tgcacgacat cggaaaccca aaggctatca 1501 agtgcgtgcc ccaggccgac gtggagtgga agttctacga cgctcagccc tgctccgaca 1561 aggcctacaa gatcgaggaa ctgttctact cttacgctac ccacagcgac aagttcaccg 1621 acggcgtgtg cctgttctgg aactgcaacg tggacaggta cccagccaac tctatcgtgt 1681 gcagattcga cacccgtgtg ctgagcaacc tgaacctgcc cggatgcgac ggcggatccc 1741 tgtacgtgaa caagcacgcc ttccacaccc ctgctttcga caagtctgcc ttcgtgaacc 1801 tgaagcagct gcctttcttc tactacagcg actccccatg cgagagccac ggcaagcagg 1861 tggtgtccga catcgactac gtgcccctga agagcgccac ctgcatcacc cgctgcaacc 1921 tgggcggcgc cgtgtgcaga caccacgcca acgaatacag gctgtacctg gacgcttaca 1981 acatgatgat cagcgccggc ttctccctgt gggtgtacaa gcagttcgac acctacaacc 2041 tgtggaacac cttcaccagg ctgcagtaat acccaacttt cttgtacaaa gttggcatta 2101 taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa 2161 tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg 2221 tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa 2281 atatatcatc atgcctcctc tagaccagcc aggacagaaa tgcctcgact tcgctgctac 2341 ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat 2401 aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca 2461 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg 2521 actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg 2581 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta 2641 aaacaaagtt aaacattatg agggaagcgg tgatcgccga agtatcgact caactatcag 2701 aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg 2761 gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga 2821 ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg 2881 cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg 2941 acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca 3001 atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc 3061 tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg 3121 atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact 3181 cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt 3241 acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc 3301 gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag 3361 aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag 3421 gcgagatcac caaggtagtc ggcaaataac cctcgagcca cccatgacca aaatccctta 3481 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt 3541 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 3601 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 3661 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact 3721 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 3781 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 3841 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 3901 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 3961 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 4021 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 4081 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 4141 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg 4201 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc 4261 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa 4321 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgaca //