LOCUS       sequence-324313-        3894 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 
            K164A/H165A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3894)
  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 3894)
  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 Delta RC
FEATURES             Location/Qualifiers
     source          1..3894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(494..499,500..516,517..522))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      1179..1767
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1668..1687
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1902..1929)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2021..2107)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    2160..2259
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2803..2902)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3014..3033)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3092..3111)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(join(3685..3894,1..324))
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 gatatagatc tcactacgcg gctgctcaaa cttgggcaga acgtaagccg cgagagcgcc
       61 aacaaccgct tcttggtcga aggcagcaag cgcgatgaat gtcttactac ggagcaagtt
      121 cccgaggtaa tcggagtccg gctgatgttg ggagtaggtg gctacgtctc cgaactcacg
      181 accgaaaaga tcaagagcag cccgcatgga tttgacttgg tcagggccga gcctacatgt
      241 gcgaatgatg cccatacttg agccacctaa ctttgtttta gggcgactgc cctgctgcgt
      301 aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt
      361 ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga aaaccgccac
      421 tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatacg
      481 ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc gtgccttcat
      541 ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg aggcatttct
      601 gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg cattggcggc
      661 cttgctgttc ttctacggca aggtgctgtg cacggatctg ccctggcttc aggagatcgg
      721 aagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag tggttcgcat
      781 cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct atagttctag
      841 tggttggcta ctaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
      901 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
      961 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     1021 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     1081 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1141 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1201 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1261 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1321 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1381 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1441 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1501 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1561 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     1621 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     1681 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     1741 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     1801 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     1861 aaccgtatta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
     1921 tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
     1981 cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     2041 cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt
     2101 ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat ctcgggcccc
     2161 aaataatgat tttattttga ctgatagtga cctgttcgtt gcaacaaatt gatgagcaat
     2221 gcttttttat aatgccaact ttgtacaaaa aagttggcat ggagagcctg gtgcctggct
     2281 tcaacgaaaa gacccacgtg cagctgtctc tgccagtgct gcaggtgcgc gacgtgctgg
     2341 tgaggggatt cggtgactct gtggaggaag tgctgtctga agctagacag cacctgaagg
     2401 atggtacctg cggactggtg gaggtggaaa agggcgtgct gccccagctg gagcagcctt
     2461 acgtgttcat caagcgcagc gatgctagga ctgctcccca cggacacgtg atggtggaac
     2521 tggtggccga gctggaaggt atccagtacg gtcgctccgg agagaccctg ggtgtgctgg
     2581 tgccacacgt gggtgaaatc cccgtggctt acagaaaggt gctgctgcgt aagaacggaa
     2641 acaagggtgc cggcggacac tcctacggtg ctgacctgaa gtctttcgac ctgggcgacg
     2701 agctgggaac tgacccttac gaggacttcc aggaaaactg gaacaccgcg gccagctccg
     2761 gagtgaccag agagctgatg cgtgaactga acggtggcta atacccaact ttcttgtaca
     2821 aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca
     2881 gtcaaaataa aatcattatt tgccatccag ctgcagctct ggcccgtgtc tcaaaatctc
     2941 tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac
     3001 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga
     3061 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
     3121 caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg
     3181 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
     3241 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     3301 tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct
     3361 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     3421 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca
     3481 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
     3541 gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc
     3601 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aggccgggaa
     3661 gccgatctcg gcttgaacga attgttaggt ggcggtactt gggtcgatat caaagtgcat
     3721 cacttcttcc cgtatgccca actttgtata gagagccact gcgggatcgt caccgtaatc
     3781 tgcttgcacg tagatcacat aagcaccaag cgcgttggcc tcatgcttga ggagattgat
     3841 gagcgcggtg gcaatgccct gcctccggtg ctcgccggag actgcgagat cata
//
