LOCUS       sequence-295491-        4941 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 4941)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4941)
  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-m4133965
FEATURES             Location/Qualifiers
     source          1..4941
                     /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             716..733
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    810..931
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(821..840)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     875..894
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1046..1092
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1122..1258
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1303..1374
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1600..2188)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1680..1699)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2934..3002,3003..3794)
                     /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(3152..3171)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3807..3943
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4095..4166
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4171..4217)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4235..4283
                     /note="synthetic polyadenylation signal"
     enhancer        4364..4667
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4669..4871
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4822..4842
                     /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 tcagtcgatg agtgacaaaa tcattcatct aaccgatgac tcctttgata cagacgtgtt
      421 gaaggccgat ggtgcgatac tggtagattt ctgggctgaa tggtgtggcc cgtgcaagat
      481 gatcgcaccc attctggacg agatcgccga cgagtatcaa ggaaaattaa ctgtcgctaa
      541 gcttaatata gatcagaacc cagggacggc acctaaatac ggcatcaggg gaatacctac
      601 cctcctgctc tttaagaatg gggaagttgc cgcgacaaaa gtgggagccc tgtctaaggg
      661 ccagctaaaa gagttcttgg acgctaacct ggcaggttca ggctcggggc acatgcacca
      721 tcaccatcac catagcagcg ggctcgtgcc acgcggctaa gctagcttga ctgactgaga
      781 tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac
      841 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      901 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
      961 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     1021 tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg
     1081 aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa
     1141 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     1201 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg
     1261 tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt
     1321 ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca
     1381 ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc
     1441 gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg
     1501 ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca
     1561 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1621 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1681 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1741 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1801 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1861 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1921 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1981 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2041 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2101 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2161 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2221 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2281 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2341 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2401 tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga
     2461 aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca
     2521 agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta
     2581 ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa
     2641 tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa
     2701 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     2761 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     2821 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     2881 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     2941 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3001 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     3061 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3121 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3181 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3241 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3301 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     3361 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3421 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3481 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     3541 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3601 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3661 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3721 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3781 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     3841 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     3901 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa
     3961 actcgactct ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc
     4021 tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg
     4081 atcgcggctc ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt
     4141 cataacagca gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc
     4201 cccaaggggt tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat
     4261 cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat
     4321 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     4381 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     4441 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     4501 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     4561 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     4621 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt
     4681 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     4741 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     4801 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     4861 taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc
     4921 gacacacccg ccagcggccg c
//
