LOCUS       sequence-297354-        6825 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6825)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6825)
  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-m4252958
FEATURES             Location/Qualifiers
     source          1..6825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             251..271
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             272..289
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    358..479
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(369..388)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     423..442
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      594..640
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      670..806
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    851..922
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1148..1736)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1228..1247)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2236..3445
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3282..3301
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3449..4477
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4504..4552
                     /note="synthetic polyadenylation signal"
     CDS             join(4818..4886,4887..5678)
                     /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(5036..5055)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5691..5827
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5979..6050
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6055..6101)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6119..6167
                     /note="synthetic polyadenylation signal"
     enhancer        6248..6551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6553..6755
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6706..6726
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaaa atgtcagatg taaagtgcac atcagtagtc ttactctcag ttttgcaaca
       61 actcagagta gaatcatcat ctaaattgtg ggctcaatgt gtccagttac acaatgacat
      121 tctcttagct aaagatacta ctgaagcctt tgaaaaaatg gtttcactac tttctgtttt
      181 gctttccatg cagggtgctg tagacataaa caagctttgt gaagaaatgc tggacaacag
      241 ggcaacctta gaaaaccttt actttcagtc gcaccatcac catcaccatt aaggatccgc
      301 aggcctctgc tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa
      361 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
      421 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
      481 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
      541 aaagcaagta aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata
      601 accccttggg gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc
      661 tatctaccgg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc
      721 cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt
      781 cctccttacc agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt
      841 tcgatcaaga cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc
      901 atcaaacttt cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa
      961 ccagtcaagt cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt
     1021 gcgtcagtca agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca
     1081 tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1141 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1201 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1261 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1321 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1381 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1441 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1501 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1561 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1621 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1681 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1741 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1801 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     1861 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     1921 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat
     1981 ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg
     2041 caggctacct ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt
     2101 gcttggcagc gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac
     2161 ccgagagaag ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga
     2221 aatcggggcg cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcg
     2281 tcacggcgag cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg
     2341 gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca
     2401 gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag
     2461 cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg
     2521 cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca
     2581 gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag
     2641 cgggctgctg ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg
     2701 tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg
     2761 ggttgggggg agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt
     2821 gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa
     2881 ggtcttgagc ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctgggcacc
     2941 atctggggac cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg
     3001 cgggggcggc agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg
     3061 ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacca
     3121 gccggtaggt gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta
     3181 ggctctcctg aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca
     3241 gtttctttgg tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac
     3301 tatgcgctcg gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta
     3361 atcatttggg tcaatatgta attttcagtg ttagacttgt aaattgtccg ctaaattctg
     3421 gccgtttttg gcttttttgt tagacaacat gggtaaaaag cctgaactca ccgcgacgtc
     3481 tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga
     3541 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt
     3601 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatccgc
     3661 cgcgctcccg attccggaag tgcttgacat tggggagttc agcgagagcc tgacctattg
     3721 catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
     3781 tgttctgcag ccggtcgcgg aggcaatgga tgccatcgct gccgccgatc ttagccagac
     3841 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
     3901 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
     3961 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
     4021 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
     4081 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
     4141 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
     4201 gcggaggcat ccggagcttg caggatcgcc ccggctccgg gcgtatatgc tccgcattgg
     4261 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
     4321 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
     4381 ccgcagaagc gccgccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
     4441 ccgacgcccc agcactcgtc cgagggcaaa ggaataagct agtatgtaag cctagtctta
     4501 gataataaaa tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc
     4561 taggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg
     4621 ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat
     4681 attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt
     4741 ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat
     4801 cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4861 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4921 aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4981 ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat
     5041 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5101 attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca
     5161 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5221 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5281 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5341 agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg
     5401 aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg
     5461 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5521 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5581 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5641 tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc
     5701 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     5761 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     5821 tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg
     5881 agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat
     5941 cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac
     6001 aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa
     6061 acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca
     6121 cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc taactgctga
     6181 tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg
     6241 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     6301 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     6361 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     6421 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     6481 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     6541 ctattaccat gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     6601 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     6661 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     6721 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat
     6781 ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgc
//
