LOCUS       sequence-295480-        6938 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 6938)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6938)
  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-m4133890
FEATURES             Location/Qualifiers
     source          1..6938
                     /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..409
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    478..599
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(489..508)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     543..562
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      714..760
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      790..926
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    971..1042
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1268..1856)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1348..1367)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2356..3565
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3402..3421
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3569..4597
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4624..4672
                     /note="synthetic polyadenylation signal"
     CDS             join(4938..5006,5007..5798)
                     /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(5156..5175)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5811..5947
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6092..6163
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6168..6214)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6232..6280
                     /note="synthetic polyadenylation signal"
     enhancer        6361..6664
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6666..6868
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6819..6839
                     /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 tcagtcgggg aaacctatac ccaatccact tctcggctta gattctacct aaggatccgc
      421 aggcctctgc tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa
      481 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
      541 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
      601 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
      661 aaagcaagta aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata
      721 accccttggg gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc
      781 tatctaccgg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc
      841 cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt
      901 cctccttacc agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt
      961 tcgatcaaga cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc
     1021 atcaaacttt cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa
     1081 ccagtcaagt cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt
     1141 gcgtcagtca agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca
     1201 tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1261 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1321 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1381 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1441 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1501 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1561 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1621 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1681 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1741 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1801 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1861 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1921 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     1981 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     2041 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat
     2101 ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg
     2161 caggctacct ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt
     2221 gcttggcagc gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac
     2281 ccgagagaag ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga
     2341 aatcggggcg cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcg
     2401 tcacggcgag cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg
     2461 gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca
     2521 gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag
     2581 cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg
     2641 cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca
     2701 gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag
     2761 cgggctgctg ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg
     2821 tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg
     2881 ggttgggggg agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt
     2941 gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa
     3001 ggtcttgagc ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctgggcacc
     3061 atctggggac cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg
     3121 cgggggcggc agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg
     3181 ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacct
     3241 gccggtaggt gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta
     3301 ggctctcctg aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca
     3361 gtttctttgg tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac
     3421 tatgcgctcg gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta
     3481 atcatttggg tcaatatgta attttcagtg ttagacttgt aaattgtccg ctaaattctg
     3541 gccgtttttg gcttttttgt tagacaacat gggtaaaaag cctgaactca ccgcgacgtc
     3601 tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga
     3661 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt
     3721 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatccgc
     3781 cgcgctcccg attccggaag tgcttgacat tggggagttc agcgagagcc tgacctattg
     3841 catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
     3901 tgttctgcag ccggtcgcgg aggcaatgga tgccatcgct gccgccgatc ttagccagac
     3961 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
     4021 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
     4081 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
     4141 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
     4201 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
     4261 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
     4321 gcggaggcat ccggagcttg caggatcgcc ccggctccgg gcgtatatgc tccgcattgg
     4381 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
     4441 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
     4501 ccgcagaagc gccgccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
     4561 ccgacgcccc agcactcgtc cgagggcaaa ggaataagct agtatgtaag cctagtctta
     4621 gataataaaa tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc
     4681 taggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg
     4741 ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat
     4801 attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt
     4861 ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat
     4921 cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4981 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5041 aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5101 ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat
     5161 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5221 attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca
     5281 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5341 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5401 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5461 agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg
     5521 aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg
     5581 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5641 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5701 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5761 tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc
     5821 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     5881 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     5941 tccttaagat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac
     6001 gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc
     6061 ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc
     6121 ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc
     6181 aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa
     6241 aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg
     6301 tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     6361 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     6421 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     6481 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     6541 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     6601 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     6661 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     6721 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     6781 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     6841 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
     6901 gatcctacca tccactcgac acacccgcca gcggccgc
//
