LOCUS       sequence-300641-        6437 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 6437)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6437)
  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-m4252813
FEATURES             Location/Qualifiers
     source          1..6437
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    94..142
                     /note="synthetic polyadenylation signal"
     terminator      160..206
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    211..282
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(434..570)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(583..1374,1375..1443))
                     /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     1206..1225
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1709..1757
                     /note="synthetic polyadenylation signal"
     CDS             complement(1784..2383)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1867..1887)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2364..2383
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2387..3596)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2531..2550)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4096..4684
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4585..4604
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4910..4981
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5026..5162)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5192..5238)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5353..5474
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5390..5409)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5444..5463
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5527..5592)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(5596..5616)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(5943..6145)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5972..5992)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(6147..6437,1..13)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
       61 ctagatctgg ctacactcga tcagcagtta gataataaaa tcgctatcca tcgaagatgg
      121 atgtgtgttg gttttttgtg tgtgtaacgc aacgattgat agcataaccc cttggggcct
      181 ctaaacgggt cttgaggggt tttttgaatt gagaggtagc tgaagctgct gttatgacca
      241 ctagagggaa gaagatacct gtggagctaa tggtccaaga tgtcgggagc cgcgatcgct
      301 gccaaaaagg taagctgacg atagctgacg agattcgatg cccgacgagc aaatgagcgc
      361 cacggctgtt cgcgtaagct cgaaacgacg gagattcgat agagccagag tcgagtttaa
      421 acgattcggg atcttaagga taaatttctg gtaaggagga cacgtatgga agtgggcaag
      481 ttggggaagc cgtatccgtt gctgaatctg gcatatgtgg gagtataaga cgcgcagcgt
      541 cgcatcaggc atttttttct gcgccaatgc acctagaatc ggttaccaat gcttaatcag
      601 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
      661 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
      721 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
      781 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatca actgttgccg
      841 ggaagctaga gtaagtagtt cgccagttaa tagtttacgc aaggttgttg ccattgctac
      901 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
      961 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1021 tccaatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1081 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgaatactc
     1141 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1201 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaagcgttc
     1261 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1321 tcgcgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1381 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1441 catgcgaaag cctcctgctg atcctgtctc ttgatagatg tttgcaaaag ctaggactcg
     1501 attatacgtt gaccgatcag actatcaata tctaggttta aacagatcgg tacgctgccg
     1561 tgaccaggct tgagtacaat atctgacgat tggacttcct gggtcccaag ctggattccg
     1621 actggctgac tgccaagcaa cgatatggat cgtcgagact cgcactgaga ggatcgttct
     1681 cggtacacca ggcgcgccta gcgttacaca cacaaaaaac caacacacat ccatcttcga
     1741 tggatagcga ttttattatc taagactagg cttacatact agcctaggca ccgggcttgc
     1801 gggtcatgca ccaggtgcgc ggtccttcgg gcacctcgac gtcggcggtg acggtgaagc
     1861 cgagccgctc gtagaagggg aggttgcggg gcgcggaggt ctccaggaag gcgggcaccc
     1921 cggcgcgctc ggcggcctcc actccgggga gcacgacggc gctgcccaga cccttgccct
     1981 ggtggtcggg cgagacgccg acggtggcca ggaaccacgc gggctccttg ggccggtgcg
     2041 gcgccaggag gccctccatc tgttgctgcg cggccagccg ggaaccgctc aactcggcca
     2101 tgcgcgggcc gatctcggcg aacaccgccc ccgcttcgac gctctccggc gtggtccaga
     2161 ccgccaccgc ggcgccgtcg tccgcgaccc acaccttgcc gatgtcgagc ccgacgcgcg
     2221 tgaggaagag ttcttgcagc tcggtgaccc gctcgatgtg gcggtccgga tcgacggtgt
     2281 ggcgcgtggc ggggtagtcg gcgaacgcgg cggcgagggt gcgtacggcc ctggggacgt
     2341 cgtcgcgggt ggcgaggcgc accgtgggct tgtactcggt catgttgtct aacaaaaaag
     2401 ccaaaaacgg ccagaattta gcggacaatt tacaagtcta acactgaaaa ttacatattg
     2461 acccaaatga ttacatttca aaaggtgcct aaaaaacttc acaaaacaca ctcgccaacc
     2521 ccgagcgcat agttcaaaac cggagcttca gctacttaag aagataggta cataaaaccg
     2581 accaaagaaa ctgacgcctc acttatccct cccctcacca gaggtccggc gcctgtcgat
     2641 tcaggagagc ctaccctagg cccgaaccct gcgtcctgcg acggagaaaa gcctaccgca
     2701 cacctaccgg ctggtggccc caccctgcat tataagccaa cagaacgggt gacgtcacga
     2761 cacgacgagg gcgcgcgctc ccaaaggtac gggtgcactg cccaacggca ccgccataac
     2821 tgccgccccc gcaacagacg acaaaccgag ttctccagtc agtgacaaac ttcacgtcag
     2881 ggtccccaga tggtgcccag cccatctcac ccgaataaga gctttcccgc attagcgaag
     2941 ggctcaagac cttgggttct tgccgcccac catgcccccc accttgtttc aacgacctca
     3001 cagcccgcct cacaagcgtc ttccattcaa gactcgggaa cagccgccat tttgctgcgc
     3061 tccccccaac ccccagttca gggcaacctt gctcgcggac ccagactaca gcccttggcg
     3121 gtctctccac acgcttccgt cccaccgagc ggcccggcgg ccacgaaagc cccggccagc
     3181 ccagcagccc gctactcacc aagtgacgat cacagcgatc cacaaacaag aaccgcgacc
     3241 caaatcccgg ctgcgacgga actagctgtg ccacacccgg cgcgtcctta tataatcatc
     3301 ggcgttcacc gccccacgga gatccctccg cagaatcgcc gagaagggac tacttttcct
     3361 cgcctgttcc gctctctgga aagaaaacca gtgccctaga gtcacccaag tcccgtccta
     3421 aaatgtcctt ctgctgatac tggggttcta aggccgagtc ttatgagcag cgggccgctg
     3481 tcctgagcgt ccgggcggaa ggatcaggac gctcctgcgc ccttcgtctg acgtggcagc
     3541 gctcgccgtg acgagggggg cgcccgcggg aggcgccaaa acccggcgcg gaggccaggc
     3601 gcgccccgat ttcgatattc ctcggatctc ggatagatcg ggattgagga tgtaggttga
     3661 acttctctcg ggtgatcccg atctcggata ttcgggattg gagtaatacc tggcgatagg
     3721 cgctgccaag caatagccaa ggcgcaaggc cggccaagag acttgcgata gttcgttcga
     3781 gaggtagcct gcaagatgga tcggacgaaa ggtttgaaga ttttccgacg agggccttgg
     3841 agagttcgga aatttaaatg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3901 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3961 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4021 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4081 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4141 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4201 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4261 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4321 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4381 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4441 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4501 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4561 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4621 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4681 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4741 catgatttaa atgaaactcg taatcggaga acgggtgcaa tagcaaggac cagatcgaac
     4801 ttgactgacg caattctgag cgtagtcgaa acccagacaa gtcgatctcg ccaagtagct
     4861 gacttgactg gttaattaag tttcgatagc ccaaggtaac caatgagatt agggagggaa
     4921 gaaagtttga tgaactactt ctgaccccta gtggtgtcca gaaaagacca ttaaaggaat
     4981 gtcttgatcg aaatcgagag atcgcctgca ggatagctga cgaccttaag gataaatttc
     5041 tggtaaggag gacacgtatg gaagtgggca agttggggaa gccgtatccg ttgctgaatc
     5101 tggcatatgt gggagtataa gacgcgcagc gtcgcatcag gcattttttt ctgcgccaat
     5161 gccggtagat agcaatccgg cccgaggggc acaaaaaacc cctcaagacc cgtttagagg
     5221 ccccaagggg ttatgctacg ataccgatag agatgggcca ataccacatt tgtagaggtt
     5281 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     5341 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5401 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5461 atcaatgtat cttatcatgt ctgtgagctg aaggtacgct gtatctcagt cagtcaagct
     5521 agcttatttg tcatcgtcgt ccttgtaatc aatgtcatga tctttgtagt ctccatcgtg
     5581 gtccttataa tccatcgact gaaagtaaag gttttcttgc agggtggctc ggttgtctaa
     5641 catctcttcg caaagtttgt tgatgtctac tgctccttgc atgctcagta gaacagatag
     5701 caagctgacc atcttctcga aagcttccgt agtatctttc gcgagcaata tatcattgtg
     5761 cagctgcacg cactgggccc agagctttga cgaactctcc acccgcagct gttgaagcac
     5821 tgataacaga actacggatg tacacttgac gtcagacatt ttgctcatgg tggcggccgc
     5881 tggcgggtgt gtcgagtgga tggtaggatc gacaaagatc tgatctgacg gttcactaaa
     5941 ccagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg
     6001 gcggagttgt tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca
     6061 ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca
     6121 ttgatgtact gccaaaaccg catcagcatg gtaatagcga tgactaatac gtagatgtac
     6181 tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta
     6241 ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag
     6301 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
     6361 actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca
     6421 ggcgggccat ttaccgt
//
