LOCUS       sequence-297429-        6389 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 6389)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6389)
  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-m4252824
FEATURES             Location/Qualifiers
     source          1..6389
                     /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    351..472
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(362..381)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     416..435
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      587..633
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      663..799
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    844..915
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1141..1729)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1221..1240)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2229..3438
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3275..3294
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3442..4041
                     /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(3442..3461)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3938..3958
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4068..4116
                     /note="synthetic polyadenylation signal"
     CDS             join(4382..4450,4451..5242)
                     /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(4600..4619)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5255..5391
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5543..5614
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5619..5665)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5683..5731
                     /note="synthetic polyadenylation signal"
     enhancer        5812..6115
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6117..6319
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6270..6290
                     /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 agctagcttg actgactgag atacagcgta ccttcagctc acagacatga taagatacat
      361 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
      421 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
      481 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
      541 gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc ataacccctt
      601 ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat tgctatctac
      661 cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
      721 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
      781 accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg atttcgatca
      841 agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag ttcatcaaac
      901 tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat taaccagtca
      961 agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga attgcgtcag
     1021 tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt tcatttaaat
     1081 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1141 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1201 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1261 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1321 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1381 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1441 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     1501 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     1561 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     1621 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     1681 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     1741 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     1801 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     1861 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     1921 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta aatttccgaa
     1981 ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc ttgcaggcta
     2041 cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct attgcttggc
     2101 agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat cacccgagag
     2161 aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat cgaaatcggg
     2221 gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc
     2281 gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc ccggacgctc
     2341 aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac
     2401 attttaggac gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca
     2461 ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa
     2521 cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga
     2581 tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg
     2641 ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag
     2701 agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact gggggttggg
     2761 gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg
     2821 gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc caaggtcttg
     2881 agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc accatctggg
     2941 gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg ttgcgggggc
     3001 ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt
     3061 cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca ccagccggta
     3121 ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc
     3181 ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg tcagtttctt
     3241 tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg aactatgcgc
     3301 tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt
     3361 gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt ctggccgttt
     3421 ttggcttttt tgttagacaa catgaccgag tacaagccca cggtgcgcct cgccacccgc
     3481 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     3541 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     3601 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     3661 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     3721 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga gggcctcctg
     3781 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     3841 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc cgccgagcgc
     3901 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     3961 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     4021 acccgcaagc ccggtgccta ggctagtatg taagcctagt cttagataat aaaatcgcta
     4081 tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt
     4141 accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc
     4201 cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg
     4261 gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta
     4321 taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg
     4381 catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4441 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4501 gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4561 cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4621 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4681 ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4741 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4801 tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4861 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4921 gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc
     4981 ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5041 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5101 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5161 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5221 ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga
     5281 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     5341 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa
     5401 tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc
     5461 cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt
     5521 tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct
     5581 ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt
     5641 ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac
     5701 acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat
     5761 ctagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     5821 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     5881 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     5941 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     6001 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     6061 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatgctgat
     6121 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     6181 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     6241 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     6301 ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt cgatcctacc
     6361 atccactcga cacacccgcc agcggccgc
//
