LOCUS       Exported                4599 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Gateway-compatible Entry vector, with insert of NSP13 gene's CDS 
            from SARS-CoV-2 isolate Wuhan-Hu-1.
ACCESSION   .
VERSION     .
KEYWORDS    pDONR223 SARS-CoV-2 NSP13
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4599)
  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 4599)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4599
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     68..84
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     68..84
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     89..108
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        89..107
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    125..224
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2034..2133)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2149..2166)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2149..2165)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2158..2180)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      2229..2315
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2407..2434
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2593..2610)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2764..3352)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2844..2863)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3445..4236)
                     /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 gcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacgggcca
       61 gagctgccag gaaacagcta tgaccatgta atacgactca ctatagggga tatcagctgg
      121 atggcaaata atgattttat tttgactgat agtgacctgt tcgttgcaac aaattgataa
      181 gcaatgcttt cttataatgc caactttgta caagaaagtt gggtattact gcagggtagc
      241 cacgttcctg cgggggatct ccagggaggt gaactgcagc ttgtcgtaca ggtctctgtc
      301 gctcatgatg cacaggattc ccactttagc tctggtgatg gccacgttga acctgttcac
      361 gttgcagctg tgggcggtct cggtggtctg ggtgaagatc acgtagtcgt attcagagcc
      421 ctgagaggag tccacggtct gggtaggcag acccaggatc ttggaagcca cggcgttctg
      481 agagttgtaa ggggagatga acacggcctt tctccaggcg gggttcctgg tcaggaactc
      541 gcgcaccaca ccgatctggg gcctgttgat ggcgctagac acgtcgtggg tgatcactcc
      601 cttgtagaac atcttgaagc actgggcgct cttgtccttg tgggccttca gcttgttgtc
      661 gtacaccaga gcagacacgg tgtccacgat ttcagcaggg cagcggcggc aggtacccag
      721 gaacatgtct ggaccgatgg tcttcatcag gcggcacacg ctgttgaagt attctggctc
      781 cagggtgccc ttggtcagca gggtacgagg agcaggcagc tgagctgggt caccgatgta
      841 cacgtagtgc ttagccctca gtctagcgtt caccacagac aggtcgtagt tggtggccat
      901 gctgatctcg tcgaacacca cgatgtcagc ggtggtttca ggcagggcgt tcacggtgca
      961 gaacacgtac tgctccaggg tagagttcac cttgaacttg tcgaagcatt ccacacgagc
     1021 tctggctggg atgatgcggg agcacttgtc gataggcagg tacttcagag ccttctcgca
     1081 cagagcatcc acagcagcgt gagaacaagc agtgtacacg atacgggcag aggggtagta
     1141 cagagccaga ccgatggcga agtgagactt gccggtacct ggaggtccct gcagggtgga
     1201 gtacttctgc atgcccactt tctggtagtt ggccacgttg gagctgaact cgtcggagat
     1261 gttcagggtt gggtacagtc cggtgatgcg cacgtagtgt tcctgaggca ccagggtggg
     1321 agcagacagt ggcatcacgg tgtgggaggt cagcacgaag tagtcaccca cgttcagctt
     1381 gtaggtggtg gttcctctgt acaccacggc gtcaccgtag tctccctttt cgaaggtgta
     1441 ctctccgatc tgcaccttgc tgttcttggt caccctgtat ccggtgaaca cgtagttacg
     1501 gttcaggggt ggtcttggct tgcccacttc ccaagacagg tgcagttcgc ggtcagacag
     1561 cacctcacgc acggtagcga taccgtaaga cagcttgaag gtttcctcgg tggccttcag
     1621 ggtctcagcg gcgaacagct tcagcctttc ggtgcaggtg ttagccagga tgtagtctcc
     1681 ggcgttggtc cagtcgcagg tagcgatggc gttgaagtcg gtcacgttgt cggagcccac
     1741 gcaggtgttc ttgtacagtc cgaacacctg gccgttagcg cacagtggga agctgatagg
     1801 gggcttgtgg gacttgcagt agtagctcat tccgcccagg tacagctggg tcacgtcagt
     1861 cacgtcacaa ccaggagcgt tgcacacgta tgggttcacg gacagcacca gcttgtggga
     1921 ggtgctgatc acgtggtcgt agcagcactt gcagcacagg aaaggcctgc ggatgcaagc
     1981 tccacatctc aggctggtct gagagttgca cagcacgcaa gcgcccacgg ccatgccaac
     2041 ttttttgtac aaagttggca ttataaaaaa gcattgctca tcaatttgtt gcaacgaaca
     2101 ggtcactatc agtcaaaata aaatcattat ttggggcccg agcttaagac tggccgtcgt
     2161 tttacaacgt cgtgactggg aaaacatcca tgctagcgtt aacgcgagag tagggaactg
     2221 ccaggcatca aataaaacga aaggctcagt cggaagactg ggcctttcgt tttatctgtt
     2281 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg
     2341 tgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa
     2401 ctaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact cttcctggct
     2461 agcggtacgc gtattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2521 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2581 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2641 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2701 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2761 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2821 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2881 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2941 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3001 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3061 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3121 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3181 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3241 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3301 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3361 tttgatcttt tctacggggt ctgacgctca gtggaacgac gcgtaactca cgttaaggga
     3421 ttttggtcat gggtggctcg agggttattt gccgactacc ttggtgatct cgcctttcac
     3481 gtagtggaca aattcttcca actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc
     3541 aagataagcc tgtctagctt caagtatgac gggctgatac tgggccggca ggcgctccat
     3601 tgcccagtcg gcagcgacat ccttcggcgc gattttgccg gttactgcgc tgtaccaaat
     3661 gcgggacaac gtaagcacta catttcgctc atcgccagcc cagtcgggcg gcgagttcca
     3721 tagcgttaag gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag
     3781 ttcctccgcc gctggaccta ccaaggcaac gctatgttct cttgcttttg tcagcaagat
     3841 agccagatca atgtcgatcg tggctggctc gaagatacct gcaagaatgt cattgcgctg
     3901 ccattctcca aattgcagtt cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg
     3961 cacaacaatg gtgacttcta cagcgcggag aatctcgctc tctccagggg aagccgaagt
     4021 ttccaaaagg tcgttgatca aagctcgccg cgttgtttca tcaagcctta cggtcaccgt
     4081 aaccagcaaa tcaatatcac tgtgtggctt caggccgcca tccactgcgg agccgtacaa
     4141 atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg acgccaacta cctctgatag
     4201 ttgagtcgat acttcggcga tcaccgcttc cctcataatg tttaactttg ttttagggcg
     4261 actgccctgc tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta
     4321 acgcgcttgc tgcttggatg cccgaggcat agactgtacc ccaaaaaaac agtcataaca
     4381 agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca
     4441 gttgcgtgag cgcatacgct acttgcatta cagcttacga accgaacagg cttatgtcca
     4501 ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct tgggtagcag
     4561 cgaagtcgag gcatttctgt cctggctggt ctagaggag
//
