LOCUS       sequence-301685-        7362 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 7362)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7362)
  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-m4252889
FEATURES             Location/Qualifiers
     source          1..7362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     33..52
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      204..250
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      280..416
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    461..532
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(758..1346)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(838..857)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1846..3055
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2892..2911
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3059..4087
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4114..4162
                     /note="synthetic polyadenylation signal"
     CDS             join(4428..4496,4497..5288)
                     /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(4646..4665)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5301..5437
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5589..5660
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5665..5711)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5729..5777
                     /note="synthetic polyadenylation signal"
     enhancer        5858..6161
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6163..6365
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6316..6336
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             6686..6706
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             6740..6817
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             6845..6865
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    join(7330..7362,1..89)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7341..7360)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
       61 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
      121 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
      181 ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag
      241 gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg
      301 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg
      361 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc
      421 gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt
      481 ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt
      541 ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga
      601 cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct
      661 attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa
      721 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
      781 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
      841 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
      901 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
      961 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1021 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1081 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1141 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     1201 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     1261 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     1321 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     1381 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     1441 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     1501 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     1561 tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa
     1621 atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag
     1681 tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact
     1741 ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc
     1801 ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggcct ccgcgccggg
     1861 ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag cgctgccacg tcagacgaag
     1921 ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg acagcggccc gctgctcata
     1981 agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg acttgggtga
     2041 ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt agtcccttct
     2101 cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat ataaggacgc
     2161 gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt
     2221 ggatcgctgt gatcgtcact tggtgagtag cgggctgctg ggctggccgg ggctttcgtg
     2281 gccgccgggc cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg
     2341 ggtccgcgag caaggttgcc ctgaactggg ggttgggggg agcgcagcaa aatggcggct
     2401 gttcccgagt cttgaatgga agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg
     2461 tggggggcat ggtgggcggc aagaacccaa ggtcttgagc ccttcgctaa tgcgggaaag
     2521 ctcttattcg ggtgagatgg gctgggcacc atctggggac cctgacgtga agtttgtcac
     2581 tgactggaga actcggtttg tcgtctgttg cgggggcggc agttatggcg gtgccgttgg
     2641 gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct
     2701 gttggcttat aatgcagggt ggggccacca gccggtaggt gtgcggtagg cttttctccg
     2761 tcgcaggacg cagggttcgg gcctagggta ggctctcctg aatcgacagg cgccggacct
     2821 ctggtgaggg gagggataag tgaggcgtca gtttctttgg tcggttttat gtacctatct
     2881 tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg gggttggcga gtgtgttttg
     2941 tgaagttttt taggcacctt ttgaaatgta atcatttggg tcaatatgta attttcagtg
     3001 ttagacttgt aaattgtccg ctaaattctg gccgtttttg gcttttttgt tagacaacat
     3061 gggtaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga
     3121 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
     3181 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
     3241 tcgttatgtt tatcggcact ttgcatccgc cgcgctcccg attccggaag tgcttgacat
     3301 tggggagttc agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt
     3361 gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggcaatgga
     3421 tgccatcgct gccgccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg
     3481 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta
     3541 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
     3601 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg
     3661 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
     3721 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc
     3781 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc
     3841 ccggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
     3901 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg
     3961 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gccgccgtct ggaccgatgg
     4021 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa
     4081 ggaataagct agtatgtaag cctagtctta gataataaaa tcgctatcca tcgaagatgg
     4141 atgtgtgttg gttttttgtg tgtgtaacgc taggcgcgcc tggtgtaccg agaacgatcc
     4201 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag
     4261 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta
     4321 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag
     4381 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac
     4441 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4501 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca
     4561 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc
     4621 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4681 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac
     4741 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4801 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg
     4861 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4921 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4981 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt
     5041 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5101 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5161 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5221 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5281 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     5341 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     5401 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga
     5461 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt
     5521 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg
     5581 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac
     5641 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag
     5701 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga
     5761 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta
     5821 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     5881 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     5941 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     6001 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     6061 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     6121 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt
     6181 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     6241 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     6301 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     6361 gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca
     6421 cccgccagcg gccgccacca tgaaaatgtc agatgtaaag tgcacatcag tagtcttact
     6481 ctcagttttg caacaactca gagtagaatc atcatctaaa ttgtgggctc aatgtgtcca
     6541 gttacacaat gacattctct tagctaaaga tactactgaa gcctttgaaa aaatggtttc
     6601 actactttct gttttgcttt ccatgcaggg tgctgtagac ataaacaagc tttgtgaaga
     6661 aatgctggac aacagggcaa ccttagaaaa cctttacttt cagtcgggac ggcgaatccc
     6721 ggggctgata aatccctgga aaaggagatg gaagaagaac tttattgcag tgtctgcggc
     6781 taaccggttc aaaaagatca gttcaagcgg cgccctagat tacgacattc ctacgaccgc
     6841 ctccgagaat ctctatttcc agggtgaatt tggcttggca caacatgacg aggccgtcga
     6901 taacaagttt aataaagaac agcagaacgc tttctatgag atccttcact taccaaatct
     6961 gaacgaagag caacgcaacg ctttcattca gagcctgaaa gacgatccct cgcagtctgc
     7021 aaatctcctg gccgaagcca agaagctgaa tgacgcacag gctccaaaag ttgataacaa
     7081 gtttaacaag gagcagcaga atgctttcta cgagatattg caccttccta atctgaatga
     7141 ggaacaaaga aatgcattta tccagtccct aaaagacgat ccatcacaga gtgccaacct
     7201 gcttgcggaa gccaaaaagc tcaatgatgc ccaggccccg aaagtagacg cgaaccatca
     7261 ataaggatcc gcaggcctct gctagcttga ctgactgaga tacagcgtac cttcagctca
     7321 cagacatgat aagatacatt gatgagtttg gacaaaccac aa
//
