LOCUS       sequence-288065-        3714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway-compatible Entry vector, with insert of NSP5 gene's CDS from
            SARS-CoV-2 isolate Wuhan-Hu-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3714)
  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 3714)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "G3 
            (Bethesda). 2020 Aug 6. pii: g3.120.401554. doi: 
            10.1534/g3.120.401554."
COMMENT     Sequence Label: pDONR223 SARS-CoV-2 NSP5
FEATURES             Location/Qualifiers
     source          1..3714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(79..106)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(198..284)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     333..355
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     347..364
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     348..364
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    380..479
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1404..1503)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1520..1539)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1521..1539)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1544..1560)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1544..1560)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1991..2782
                     /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      2875..3463
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3364..3383
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3617..3634
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga
       61 agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc
      121 ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc
      181 aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
      241 aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc
      301 tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc
      361 cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga cctgttcgtt
      421 gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa aagttggcat
      481 gtccggattc cgcaagatgg ccttcccttc tggcaaggtg gagggctgca tggtgcaggt
      541 gacctgcgga accaccaccc tgaacggtct gtggctggac gacgtggtgt actgccccag
      601 gcacgtgatc tgcacctctg aggacatgct gaaccctaac tacgaagacc tgctgatcag
      661 aaagagcaac cacaacttcc tggtgcaggc cggtaacgtg cagctgcgtg tgatcggcca
      721 ctctatgcag aactgcgtgc tgaagctgaa ggtggacacc gctaacccca agacccctaa
      781 gtacaagttc gtgagaatcc agcctggtca gaccttctcc gtgctggcct gctacaacgg
      841 aagcccatcc ggcgtgtacc agtgcgctat gcgtcccaac ttcaccatca agggcagctt
      901 cctgaacggc tcttgcggaa gcgtgggttt caacatcgac tacgactgcg tgtccttctg
      961 ctacatgcac cacatggagc tgccaactgg agtgcacgct ggaactgacc tggagggtaa
     1021 cttctacggc cccttcgtgg acagacagac tgctcaggct gctggcaccg acaccaccat
     1081 caccgtgaac gtgctggctt ggctgtacgc cgctgtgatc aacggagacc gctggttcct
     1141 gaacaggttc accaccaccc tgaacgactt caacctggtg gctatgaagt acaactacga
     1201 gcctctgacc caggaccacg tggatatcct gggtccactg agcgcccaga ctggaatcgc
     1261 tgtgctggac atgtgcgcct ccctgaagga actgctgcag aacggcatga acggacgcac
     1321 catcctggga tccgctctgc tggaggacga attcaccccc ttcgacgtgg tgaggcagtg
     1381 ctctggtgtg accttccagt aatacccaac tttcttgtac aaagttggca ttataagaaa
     1441 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     1501 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     1561 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     1621 atcatgcctc ctctagacca gccaggacag aaatgcctcg acttcgctgc tacccaaggt
     1681 tgccgggtga cgcacaccgt ggaaacggat gaaggcacga acccagtgga cataagcctg
     1741 ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca cgcaactggt ccagaacctt
     1801 gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt
     1861 ttttggggta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg
     1921 atgtttgatg ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa
     1981 gttaaacatt atgagggaag cggtgatcgc cgaagtatcg actcaactat cagaggtagt
     2041 tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt acggctccgc
     2101 agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg tgaccgtaag
     2161 gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt cggcttcccc
     2221 tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg acgacatcat
     2281 tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc gcaatgacat
     2341 tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct tgctgacaaa
     2401 agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct ttgatccggt
     2461 tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga actcgccgcc
     2521 cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc
     2581 agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc
     2641 ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac aagaagaaga
     2701 tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga aaggcgagat
     2761 caccaaggta gtcggcaaat aaccctcgag ccacccatga ccaaaatccc ttaacgtgag
     2821 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2881 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2941 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3001 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3061 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3121 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3181 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3241 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3301 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3361 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3421 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3481 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3541 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3601 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     3661 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtt
//
