LOCUS       sequence-297406-        6834 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 6834)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6834)
  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-m4252924
FEATURES             Location/Qualifiers
     source          1..6834
                     /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..298
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    367..488
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(378..397)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     432..451
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      603..649
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      679..815
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    860..931
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1157..1745)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1237..1256)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2245..3454
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3291..3310
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3458..4486
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4513..4561
                     /note="synthetic polyadenylation signal"
     CDS             join(4827..4895,4896..5687)
                     /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(5045..5064)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5700..5836
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5988..6059
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6064..6110)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6128..6176
                     /note="synthetic polyadenylation signal"
     enhancer        6257..6560
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6562..6764
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6715..6735
                     /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 gtacccctat gacgtaccag attatgcgta
      301 aggatccgca ggcctctgct agcttgactg actgagatac agcgtacctt cagctcacag
      361 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
      421 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
      481 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
      541 aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat
      601 cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg
      661 ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc
      721 ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc
      781 catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat
      841 ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag
      901 aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa
      961 cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg
     1021 ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta
     1081 cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1141 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1201 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1261 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1321 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1381 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1441 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1501 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1561 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     1621 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1681 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1741 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1801 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1861 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     1921 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1981 catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga
     2041 tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc
     2101 ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat
     2161 cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag
     2221 gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg
     2281 cccccctcgt cacggcgagc gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc
     2341 ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct tagaacccca
     2401 gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac tggttttctt
     2461 tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg cggagggatc
     2521 tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt
     2581 tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt
     2641 ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc gctcggtggg
     2701 acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc
     2761 tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa
     2821 gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca
     2881 agaacccaag gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg gtgagatggg
     2941 ctgggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt
     3001 cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg
     3061 gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg
     3121 gggccaccag ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg
     3181 cctagggtag gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt
     3241 gaggcgtcag tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg
     3301 gttttgaact atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt
     3361 tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagacttgta aattgtccgc
     3421 taaattctgg ccgtttttgg cttttttgtt agacaacatg ggtaaaaagc ctgaactcac
     3481 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     3541 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
     3601 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
     3661 tgcatccgcc gcgctcccga ttccggaagt gcttgacatt ggggagttca gcgagagcct
     3721 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
     3781 actgcccgct gttctgcagc cggtcgcgga ggcaatggat gccatcgctg ccgccgatct
     3841 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     3901 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     3961 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     4021 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     4081 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     4141 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     4201 ctacttcgag cggaggcatc cggagcttgc aggatcgccc cggctccggg cgtatatgct
     4261 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     4321 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     4381 acaaatcgcc cgcagaagcg ccgccgtctg gaccgatggc tgtgtagaag tactcgccga
     4441 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaataagcta gtatgtaagc
     4501 ctagtcttag ataataaaat cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt
     4561 gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga
     4621 tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa
     4681 tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat
     4741 attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag
     4801 agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc
     4861 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4921 aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg
     4981 gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag
     5041 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     5101 gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta
     5161 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5221 cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca
     5281 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5341 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat
     5401 taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg
     5461 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5521 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     5581 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5641 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg
     5701 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     5761 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     5821 agaaatttat ccttaagatc ccgaatcgtt taaactcgac tctggctcta tcgaatctcc
     5881 gtcgtttcga gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc
     5941 gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat
     6001 tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca
     6061 attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg
     6121 cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct
     6181 aactgctgat cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc
     6241 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     6301 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     6361 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     6421 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     6481 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     6541 tagtcatcgc tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc
     6601 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     6661 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     6721 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     6781 agatcagatc tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgc
//
