LOCUS       sequence-295753-        6908 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 6908)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6908)
  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-m4133976
FEATURES             Location/Qualifiers
     source          1..6908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             341..361
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             362..379
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    448..569
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(459..478)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     513..532
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      684..730
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      760..896
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    941..1012
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1238..1826)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1318..1337)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2326..3535
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3372..3391
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3539..4567
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4594..4642
                     /note="synthetic polyadenylation signal"
     CDS             join(4908..4976,4977..5768)
                     /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(5126..5145)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5781..5917
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6062..6133
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6138..6184)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6202..6250
                     /note="synthetic polyadenylation signal"
     enhancer        6331..6634
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6636..6838
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6789..6809
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaat gagcttagtc ctgttgcact acgacagatg tcttgtgctg ccggtactac
       61 acaaactgct tgcactgatg acaatgcgtt agcttactac aacacaacaa agggaggtag
      121 gtttgtactt gcactgttat ccgatttaca ggatttgaaa tgggctagat tccctaagag
      181 tgatggaact ggtactatct atacagaact ggaaccacct tgtaggtttg ttacagacac
      241 acctaaaggt cctaaagtga agtatttata ctttattaaa ggattaaaca acctaaatag
      301 aggtatggta cttggtagtt tagctgccac agtacgtcta gaaaaccttt actttcagtc
      361 gcaccatcac catcaccatt aaggatccgc aggcctctgc tagcttgact gactgagata
      421 cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa
      481 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
      541 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
      601 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
      661 ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag
      721 gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg
      781 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg
      841 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc
      901 gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt
      961 ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt
     1021 ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga
     1081 cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct
     1141 attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa
     1201 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     1261 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     1321 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1381 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     1441 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1501 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1561 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1621 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     1681 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     1741 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     1801 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     1861 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     1921 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     1981 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     2041 tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa
     2101 atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag
     2161 tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact
     2221 ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc
     2281 ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggcct ccgcgccggg
     2341 ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag cgctgccacg tcagacgaag
     2401 ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg acagcggccc gctgctcata
     2461 agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg acttgggtga
     2521 ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt agtcccttct
     2581 cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat ataaggacgc
     2641 gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt
     2701 ggatcgctgt gatcgtcact tggtgagtag cgggctgctg ggctggccgg ggctttcgtg
     2761 gccgccgggc cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg
     2821 ggtccgcgag caaggttgcc ctgaactggg ggttgggggg agcgcagcaa aatggcggct
     2881 gttcccgagt cttgaatgga agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg
     2941 tggggggcat ggtgggcggc aagaacccaa ggtcttgagc ccttcgctaa tgcgggaaag
     3001 ctcttattcg ggtgagatgg gctgggcacc atctggggac cctgacgtga agtttgtcac
     3061 tgactggaga actcggtttg tcgtctgttg cgggggcggc agttatggcg gtgccgttgg
     3121 gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct
     3181 gttggcttat aatgcagggt ggggccacct gccggtaggt gtgcggtagg cttttctccg
     3241 tcgcaggacg cagggttcgg gcctagggta ggctctcctg aatcgacagg cgccggacct
     3301 ctggtgaggg gagggataag tgaggcgtca gtttctttgg tcggttttat gtacctatct
     3361 tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg gggttggcga gtgtgttttg
     3421 tgaagttttt taggcacctt ttgaaatgta atcatttggg tcaatatgta attttcagtg
     3481 ttagacttgt aaattgtccg ctaaattctg gccgtttttg gcttttttgt tagacaacat
     3541 gggtaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga
     3601 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
     3661 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
     3721 tcgttatgtt tatcggcact ttgcatccgc cgcgctcccg attccggaag tgcttgacat
     3781 tggggagttc agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt
     3841 gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggcaatgga
     3901 tgccatcgct gccgccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg
     3961 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta
     4021 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
     4081 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg
     4141 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
     4201 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc
     4261 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc
     4321 ccggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
     4381 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg
     4441 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gccgccgtct ggaccgatgg
     4501 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa
     4561 ggaataagct agtatgtaag cctagtctta gataataaaa tcgctatcca tcgaagatgg
     4621 atgtgtgttg gttttttgtg tgtgtaacgc taggcgcgcc tggtgtaccg agaacgatcc
     4681 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag
     4741 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta
     4801 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag
     4861 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac
     4921 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4981 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca
     5041 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc
     5101 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5161 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac
     5221 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5281 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg
     5341 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5401 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5461 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt
     5521 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5581 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5641 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5701 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5761 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     5821 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     5881 tccatacgtg tcctccttac cagaaattta tccttaagat cgtttaaact cgactctggc
     5941 tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc
     6001 gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg
     6061 acatcttgga ccattagctc cacaggtatc ttcttccctc tagtggtcat aacagcagct
     6121 tcagctacct ctcaattcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat
     6181 gctatcaatc gttgcgttac acacacaaaa aaccaacaca catccatctt cgatggatag
     6241 cgattttatt atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc
     6301 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     6361 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     6421 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     6481 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     6541 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     6601 gtacatctac gtattagtca tcgctattac catgctgatg cggttttggc agtacatcaa
     6661 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     6721 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     6781 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
     6841 tttagtgaac cgtcagatca gatctttgtc gatcctacca tccactcgac acacccgcca
     6901 gcggccgc
//
