LOCUS       sequence-323651-        3913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway-compatible Entry vector, with insert of mutated NSP1 CDS 
            from SARS-CoV-2 isolate Wuhan-Hu-1 (amino acid substitutions 
            R124A/K125A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3913)
  AUTHORS   Banerjee AK, Blanco MR, Bruce EA, Honson DD, Chen LM, Chow A, Bhat 
            P, Ollikainen N, Quinodoz SA, Loney C, Thai J, Miller ZD, Lin AE, 
            Schmidt MM, Stewart DG, Goldfarb D, De Lorenzo G, Rihn SJ, Voorhees 
            RM, Botten JW, Majumdar D, Guttman M
  TITLE     SARS-CoV-2 Disrupts Splicing, Translation, and Protein Trafficking 
            to Suppress Host Defenses.
  JOURNAL   Cell. 2020 Nov 25;183(5):1325-1339.e21. doi: 
            10.1016/j.cell.2020.10.004. Epub 2020 Oct 8.
  PUBMED    33080218
REFERENCE   2  (bases 1 to 3913)
  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"; doi: 
            "10.1016/j.cell.2020.10.004"; journalName: "Cell"; date: 
            "2020-11-25- 25"; volume: "183"; issue: "5"; pages: "1325-1339.e21"
COMMENT     Sequence Label: pDONR207 SARS-CoV-2 NSP1 124A/125A
FEATURES             Location/Qualifiers
     source          1..3913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(517..522,523..539,540..545))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      1202..1790
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1691..1710
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1925..1952)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2044..2130)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    2183..2282
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2826..2925)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3037..3056)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3115..3134)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(join(3708..3913,1..328))
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 catagatata gatctcacta cgcggctgct caaacttggg cagaacgtaa gccgcgagag
       61 cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat gaatgtctta ctacggagca
      121 agttcccgag gtaatcggag tccggctgat gttgggagta ggtggctacg tctccgaact
      181 cacgaccgaa aagatcaaga gcagcccgca tggatttgac ttggtcaggg ccgagcctac
      241 atgtgcgaat gatgcccata cttgagccac ctaactttgt tttagggcga ctgccctgct
      301 gcgtaacatc gttgctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc
      361 acggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacaaa aaaacagtca
      421 taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt caaggttctg
      481 gaccagttgc gtgagcgcat acgctacttg cattacagtt tacgaaccga acaggcttat
      541 gtcaactggg ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc aaccttgggc
      601 agcagcgaag tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt ttcggtctcc
      661 acgcatcgtc aggcattggc ggccttgctg ttcttctacg gcaaggtgct gtgcacggat
      721 ctgccctggc ttcaggagat cggaagacct cggccgtcgc ggcgcttgcc ggtggtgctg
      781 accccggatg aagtggttcg catcctcggt tttctggaag gcgagcatcg tttgttcgcc
      841 caggactcta gctatagttc tagtggttgg ctactaatag gttgtattga tgttggacga
      901 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
      961 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     1021 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     1081 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     1141 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     1201 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     1261 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     1321 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     1381 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     1441 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     1501 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     1561 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     1621 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     1681 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     1741 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     1801 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     1861 cgttatcccc tgattctgtg gataaccgta ttaccgctag ccaggaagag tttgtagaaa
     1921 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
     1981 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
     2041 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     2101 gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa
     2161 cgctagcatg gatctcgggc cccaaataat gattttattt tgactgatag tgacctgttc
     2221 gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca aaaaagttgg
     2281 catggagagc ctggtgcctg gcttcaacga aaagacccac gtgcagctgt ctctgccagt
     2341 gctgcaggtg cgcgacgtgc tggtgagggg attcggtgac tctgtggagg aagtgctgtc
     2401 tgaagctaga cagcacctga aggatggtac ctgcggactg gtggaggtgg aaaagggcgt
     2461 gctgccccag ctggagcagc cttacgtgtt catcaagcgc agcgatgcta ggactgctcc
     2521 ccacggacac gtgatggtgg aactggtggc cgagctggaa ggtatccagt acggtcgctc
     2581 cggagagacc ctgggtgtgc tggtgccaca cgtgggtgaa atccccgtgg cttacagaaa
     2641 ggtgctgctg gctgccaacg gaaacaaggg tgccggcgga cactcctacg gtgctgacct
     2701 gaagtctttc gacctgggcg acgagctggg aactgaccct tacgaggact tccaggaaaa
     2761 ctggaacacc aagcacagct ccggagtgac cagagagctg atgcgtgaac tgaacggtgg
     2821 ctaataccca actttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     2881 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgcagc
     2941 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     3001 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     3061 aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     3121 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc
     3181 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     3241 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     3301 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat
     3361 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     3421 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     3481 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     3541 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc
     3601 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     3661 acgaggggaa attaggccgg gaagccgatc tcggcttgaa cgaattgtta ggtggcggta
     3721 cttgggtcga tatcaaagtg catcacttct tcccgtatgc ccaactttgt atagagagcc
     3781 actgcgggat cgtcaccgta atctgcttgc acgtagatca cataagcacc aagcgcgttg
     3841 gcctcatgct tgaggagatt gatgagcgcg gtggcaatgc cctgcctccg gtgctcgccg
     3901 gagactgcga gat
//
