LOCUS       sequence-296395-        4707 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. Does not contain stop codon. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4707)
  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 4707)
  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_nostop
FEATURES             Location/Qualifiers
     source          1..4707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     359..376
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(535..562)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(654..740)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     789..811
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     803..820
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     804..820
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    836..935
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2853..2952)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2969..2988)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2970..2988)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2993..3009)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2993..3009)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3440..4231
                     /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      join(4324..4707,1..205)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
       61 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
      121 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
      181 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
      241 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
      301 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
      361 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
      421 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
      481 gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg
      541 ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct
      601 gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc
      661 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac
      721 tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg
      781 gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccccaaat
      841 aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgatg agcaatgctt
      901 ttttataatg ccaactttgt acaaaaaagt tggcatggct tacacccgct acgtggacaa
      961 caacttctgc ggtcccgacg gctaccctct ggaatgcatc aaggacctgc tggctagagc
     1021 tggcaaggct agctgcaccc tgtccgagca gctggacttc atcgacacca agagaggagt
     1081 gtactgctgc cgtgagcacg aacacgagat cgcttggtac accgaaagat ccgagaagtc
     1141 ttacgagctg cagaccccat tcgaaatcaa gctggccaag aagttcgaca ccttcaacgg
     1201 cgagtgccca aacttcgtgt tccccctgaa cagcatcatc aagaccatcc agccccgtgt
     1261 ggagaagaag aagctggacg gattcatggg tcgcatccgc agcgtgtacc cagtggctag
     1321 ccccaacgaa tgcaaccaga tgtgcctgtc taccctgatg aagtgcgacc actgcggaga
     1381 gaccagctgg cagaccggtg acttcgtgaa ggctacctgc gaattctgcg gaaccgagaa
     1441 cctgaccaag gaaggagcta ccacctgtgg atacctgcca cagaacgccg tggtgaagat
     1501 ctactgcccc gcttgccaca actccgaagt gggccctgag cactctctgg ccgaatacca
     1561 caacgagtcc ggactgaaga ccatcctgcg caagggcgga aggaccatcg ctttcggtgg
     1621 ctgcgtgttc tcttacgtgg gctgccacaa caagtgtgct tactgggtgc ctagagcttc
     1681 cgctaacatc ggttgcaacc acaccggcgt ggtgggcgaa ggatctgagg gtctgaacga
     1741 caacctgctg gagatcctgc agaaggaaaa ggtgaacatc aacatcgtgg gcgacttcaa
     1801 gctgaacgag gaaatcgcca tcatcctggc tagcttctcc gccagcacca gcgccttcgt
     1861 ggaaaccgtg aagggtctgg actacaaggc tttcaagcag atcgtggagt cctgcggcaa
     1921 cttcaaggtg accaagggaa aggccaagaa gggtgcttgg aacatcggcg agcagaagag
     1981 catcctgtcc ccactgtacg ccttcgcttc tgaagccgct agagtggtgc gttctatctt
     2041 cagcagaacc ctggagaccg cccagaacag cgtgcgtgtg ctgcagaagg ccgctatcac
     2101 catcctggac ggaatctccc agtactctct gcgcctgatc gacgccatga tgttcaccag
     2161 cgacctggct accaacaacc tggtggtcat ggcctacatc accggaggtg tggtgcagct
     2221 gacctcccag tggctgacca acatcttcgg taccgtgtac gagaagctga agcctgtgct
     2281 ggactggctg gaggaaaagt tcaaggaagg cgtggagttc ctgagggacg gatgggagat
     2341 cgtgaagttc atctctacct gcgcttgcga aatcgtgggc ggacagatcg tgacctgcgc
     2401 caaggaaatc aaggagagcg tgcagacctt cttcaagctg gtgaacaagt tcctggccct
     2461 gtgcgctgac tccatcatca tcggtggcgc caagctgaag gctctgaacc tgggagagac
     2521 cttcgtgacc cacagcaagg gtctgtaccg caagtgcgtg aagtccaggg aggaaaccgg
     2581 actgctgatg cctctgaagg ccccaaagga gatcatcttc ctggaaggag agaccctgcc
     2641 taccgaagtg ctgaccgagg aagtggtgct gaagactggc gacctgcagc cactggagca
     2701 gcctacctcc gaagccgtgg aggctcctct ggtgggtacc ccagtgtgca tcaacggcct
     2761 gatgctgctg gagatcaagg acaccgaaaa gtactgcgcc ctggctccaa acatgatggt
     2821 gaccaacaac accttcaccc tgaagggagg ttacccaact ttcttgtaca aagttggcat
     2881 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     2941 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     3001 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     3061 aaatatatca tcatgcctcc tctagaccag ccaggacaga aatgcctcga cttcgctgct
     3121 acccaaggtt gccgggtgac gcacaccgtg gaaacggatg aaggcacgaa cccagtggac
     3181 ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac gcaactggtc
     3241 cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta
     3301 tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc
     3361 cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg gcagtcgccc
     3421 taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc
     3481 agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta
     3541 cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc tggttacggt
     3601 gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc
     3661 ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga
     3721 cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg
     3781 caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt
     3841 gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt
     3901 tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa
     3961 ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg
     4021 gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga
     4081 gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca
     4141 agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa
     4201 aggcgagatc accaaggtag tcggcaaata accctcgagc cacccatgac caaaatccct
     4261 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4321 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4381 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4441 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4501 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4561 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4621 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4681 gacctacacc gaactgagat acctaca
//
