LOCUS       sequence-307274-        4578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4578)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4578)
  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"
COMMENT     Sequence Label: pGBW-m4134442
FEATURES             Location/Qualifiers
     source          1..4578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             347..367
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             368..394
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    447..568
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(458..477)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     512..531
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      683..729
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      759..895
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    940..1011
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1237..1825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1317..1336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2571..2639,2640..3431)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2789..2808)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3444..3580
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3732..3803
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3808..3854)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3872..3920
                     /note="synthetic polyadenylation signal"
     enhancer        4001..4304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4306..4508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4459..4479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaat aacgagttaa gcccagtagc gcttcgccag atgagttgcg ccgcaggtac
       61 aacccaaact gcttgtacgg atgacaacgc cctcgcttac tataatacca caaagggagg
      121 ccgattcgtt ctggccctgt tgtcggatct gcaagacttg aaatgggcac gttttcccaa
      181 gtccgacggg actggcacca tttacacaga actagagccg ccatgcaggt tcgtgactga
      241 tacccctaaa ggacccaagg tcaaatatct gtattttatc aaggggctta acaatctcaa
      301 tagaggcatg gtgttaggat ctctagccgc aacggtgcgg ctgcaagaaa acctttactt
      361 tcagtcgtac ccctatgacg taccagatta tgcgtaagct agcttgactg actgagatac
      421 agcgtacctt cagctcacag acatgataag atacattgat gagtttggac aaaccacaac
      481 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
      541 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
      601 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
      661 tggcccatct ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg
      721 ggttttttgt gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc
      781 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
      841 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg
      901 tcagctatcc tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc
      961 tggacaccac taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg
     1021 gttaccttgg gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac
     1081 ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta
     1141 ttgcacccgt tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa
     1201 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1261 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1321 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1381 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1441 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1501 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1561 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1621 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1681 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1741 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1801 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1861 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1921 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1981 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2041 atttcgttca tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa
     2101 tcttcaaacc tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt
     2161 ctcttggccg gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc
     2221 caatcccgaa tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc
     2281 cgatctatcc gagatccgag gaatatcgaa atcggggcgc gcctggtgta ccgagaacga
     2341 tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc
     2401 cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc
     2461 gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc
     2521 tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc
     2581 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2641 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt
     2701 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct
     2761 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     2821 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt
     2881 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2941 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga
     3001 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3061 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3121 tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3181 agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3241 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3301 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3361 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3421 agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc
     3481 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
     3541 acttccatac gtgtcctcct taccagaaat ttatccttaa gatcccgaat cgtttaaact
     3601 cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca
     3661 tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc
     3721 gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc tagtggtcat
     3781 aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt tagaggcccc
     3841 aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca catccatctt
     3901 cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc tagtaatcaa
     3961 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     4021 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     4081 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     4141 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     4201 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     4261 ctacttggca gtacatctac gtattagtca tcgctattac catgctgatg cggttttggc
     4321 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     4381 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     4441 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     4501 gcagagctgg tttagtgaac cgtcagatca gatctttgtc gatcctacca tccactcgac
     4561 acacccgcca gcggccgc
//
