LOCUS       sequence-296484-        4710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway-compatible Entry vector, with insert of NSP2 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 4710)
  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 4710)
  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 NSP2
FEATURES             Location/Qualifiers
     source          1..4710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(113..140)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(232..318)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     367..389
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     381..398
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     382..398
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    414..513
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2434..2533)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2550..2569)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2551..2569)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2574..2590)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2574..2590)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3021..3812
                     /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      3905..4493
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4394..4413
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4647..4664
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
       61 gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc
      121 atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc
      181 ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct
      241 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg
      301 agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga
      361 tgttttccca gtcacgacgt tgtaaaacga cggccagtct taagctcggg ccccaaataa
      421 tgattttatt ttgactgata gtgacctgtt cgttgcaaca aattgatgag caatgctttt
      481 ttataatgcc aactttgtac aaaaaagttg gcatggctta cacccgctac gtggacaaca
      541 acttctgcgg tcccgacggc taccctctgg aatgcatcaa ggacctgctg gctagagctg
      601 gcaaggctag ctgcaccctg tccgagcagc tggacttcat cgacaccaag agaggagtgt
      661 actgctgccg tgagcacgaa cacgagatcg cttggtacac cgaaagatcc gagaagtctt
      721 acgagctgca gaccccattc gaaatcaagc tggccaagaa gttcgacacc ttcaacggcg
      781 agtgcccaaa cttcgtgttc cccctgaaca gcatcatcaa gaccatccag ccccgtgtgg
      841 agaagaagaa gctggacgga ttcatgggtc gcatccgcag cgtgtaccca gtggctagcc
      901 ccaacgaatg caaccagatg tgcctgtcta ccctgatgaa gtgcgaccac tgcggagaga
      961 ccagctggca gaccggtgac ttcgtgaagg ctacctgcga attctgcgga accgagaacc
     1021 tgaccaagga aggagctacc acctgtggat acctgccaca gaacgccgtg gtgaagatct
     1081 actgccccgc ttgccacaac tccgaagtgg gccctgagca ctctctggcc gaataccaca
     1141 acgagtccgg actgaagacc atcctgcgca agggcggaag gaccatcgct ttcggtggct
     1201 gcgtgttctc ttacgtgggc tgccacaaca agtgtgctta ctgggtgcct agagcttccg
     1261 ctaacatcgg ttgcaaccac accggcgtgg tgggcgaagg atctgagggt ctgaacgaca
     1321 acctgctgga gatcctgcag aaggaaaagg tgaacatcaa catcgtgggc gacttcaagc
     1381 tgaacgagga aatcgccatc atcctggcta gcttctccgc cagcaccagc gccttcgtgg
     1441 aaaccgtgaa gggtctggac tacaaggctt tcaagcagat cgtggagtcc tgcggcaact
     1501 tcaaggtgac caagggaaag gccaagaagg gtgcttggaa catcggcgag cagaagagca
     1561 tcctgtcccc actgtacgcc ttcgcttctg aagccgctag agtggtgcgt tctatcttca
     1621 gcagaaccct ggagaccgcc cagaacagcg tgcgtgtgct gcagaaggcc gctatcacca
     1681 tcctggacgg aatctcccag tactctctgc gcctgatcga cgccatgatg ttcaccagcg
     1741 acctggctac caacaacctg gtggtcatgg cctacatcac cggaggtgtg gtgcagctga
     1801 cctcccagtg gctgaccaac atcttcggta ccgtgtacga gaagctgaag cctgtgctgg
     1861 actggctgga ggaaaagttc aaggaaggcg tggagttcct gagggacgga tgggagatcg
     1921 tgaagttcat ctctacctgc gcttgcgaaa tcgtgggcgg acagatcgtg acctgcgcca
     1981 aggaaatcaa ggagagcgtg cagaccttct tcaagctggt gaacaagttc ctggccctgt
     2041 gcgctgactc catcatcatc ggtggcgcca agctgaaggc tctgaacctg ggagagacct
     2101 tcgtgaccca cagcaagggt ctgtaccgca agtgcgtgaa gtccagggag gaaaccggac
     2161 tgctgatgcc tctgaaggcc ccaaaggaga tcatcttcct ggaaggagag accctgccta
     2221 ccgaagtgct gaccgaggaa gtggtgctga agactggcga cctgcagcca ctggagcagc
     2281 ctacctccga agccgtggag gctcctctgg tgggtacccc agtgtgcatc aacggcctga
     2341 tgctgctgga gatcaaggac accgaaaagt actgcgccct ggctccaaac atgatggtga
     2401 ccaacaacac cttcaccctg aagggaggtt aatacccaac tttcttgtac aaagttggca
     2461 ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
     2521 aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag
     2581 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     2641 aaaatatatc atcatgcctc ctctagacca gccaggacag aaatgcctcg acttcgctgc
     2701 tacccaaggt tgccgggtga cgcacaccgt ggaaacggat gaaggcacga acccagtgga
     2761 cataagcctg ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca cgcaactggt
     2821 ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt
     2881 atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg
     2941 ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc
     3001 ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc cgaagtatcg actcaactat
     3061 cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt
     3121 acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg
     3181 tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt
     3241 cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg
     3301 acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc
     3361 gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct
     3421 tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct
     3481 ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga
     3541 actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt
     3601 ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg
     3661 agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac
     3721 aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga
     3781 aaggcgagat caccaaggta gtcggcaaat aaccctcgag ccacccatga ccaaaatccc
     3841 ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3901 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3961 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4021 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4081 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4141 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4201 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4261 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4321 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4381 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4441 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4501 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4561 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4621 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     4681 caatacgcaa accgcctctc cccgcgcgtt
//
