LOCUS       Exported                3390 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Gateway-compatible Entry vector, with insert of NSP8 gene's CDS from
            SARS-CoV-2 isolate Wuhan-Hu-1.
ACCESSION   .
VERSION     .
KEYWORDS    pDONR223 SARS-CoV-2 NSP8
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3390)
  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 3390)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3390
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     158..174
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     158..174
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     179..198
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        179..197
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    215..314
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(915..1014)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1030..1047)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1030..1046)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1039..1061)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      1110..1196
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1288..1315
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1474..1491)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1645..2233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1725..1744)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2326..3117)
                     /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"
ORIGIN
        1 tgccttcatc cgtttccacg gtgtgcgtca cccggcaacc ttgggtagca gcgaagtcga
       61 ggcatttctg tcctggctgg tctagaggag gcatgatgat atatttttat cttgtgcaat
      121 gtaacatcag agattttgag acacgggcca gagctgccag gaaacagcta tgaccatgta
      181 atacgactca ctatagggga tatcagctgg atggcaaata atgattttat tttgactgat
      241 agtgacctgt tcgttgcaac aaattgataa gcaatgcttt cttataatgc caactttgta
      301 caagaaagtt gggtattact gcagcttcac ggcagagtta gcgcgcaggg cggtcacgat
      361 cagtggccaa gccaggtttg ggctgttgtc catgctgatt tcagacagct gcacgatctt
      421 agagtcagcg tccaccacct gctggatctc ccacagagcg gaggcgtagg tgaaggtggt
      481 tccgtcgcag gtgttcttgt aggtgttgta gtcggggatg accaccatca gcttagcggc
      541 ggtggtcaga gggatgatgt tcaggggcac gcagccgtct ctggcgttgt tgatgatgtt
      601 gttcagagcg tcgttgtcca gcttgcgcag catggtgaac agcatggtct gcatggcgct
      661 ggtcacctta gccctcttgt cctcagagcg agcctgcttg tacatctggg tcatagcctg
      721 gtcggccatc ttttccagct tacgctgcat agcggcgtct ctgtcgaact cagacttggc
      781 cacgttcagg gacttcttca gcttcttcag caccacttcg gagtcgccgt tagccacggc
      841 ctgttcgtaa gcctcctgag cggtggcgaa agcggcgtag gaaggcaggg agctgaactc
      901 gctagcgatg gccatgccaa cttttttgta caaagttggc attataaaaa agcattgctc
      961 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttggggccc
     1021 gagcttaaga ctggccgtcg ttttacaacg tcgtgactgg gaaaacatcc atgctagcgt
     1081 taacgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcggaagact
     1141 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     1201 cgggagcgga tttgaacgtt gtgaagcaac ggcccggagg gtggcgggca ggacgcccgc
     1261 cataaactgc caggcatcaa actaagcaga aggccatcct gacggatggc ctttttgcgt
     1321 ttctacaaac tcttcctggc tagcggtacg cgtattaatt gcgttgcgct cactgcccgc
     1381 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     1441 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     1501 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     1561 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1621 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1681 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1741 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1801 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1861 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1921 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1981 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2041 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2101 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2161 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2221 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2281 cgcgtaactc acgttaaggg attttggtca tgggtggctc gagggttatt tgccgactac
     2341 cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg cgcgcgaggc
     2401 caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata
     2461 ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc
     2521 ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc
     2581 ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg
     2641 ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc
     2701 tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc
     2761 tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg
     2821 ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct
     2881 ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc
     2941 atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc
     3001 atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat
     3061 gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcataat
     3121 gtttaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tccataacat
     3181 caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac
     3241 cccaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac caccgctgcg
     3301 ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt acagcttacg
     3361 aaccgaacag gcttatgtcc actgggttcg
//
