LOCUS       sequence-354355-        6740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6740)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6740)
  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-m4134491
FEATURES             Location/Qualifiers
     source          1..6740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1180..1200
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1201..1227
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1280..1401
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1291..1310)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1345..1364
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1516..1562
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1592..1728
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1773..1844
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2070..2658)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2150..2169)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3158..4367
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4204..4223
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4371..4970
                     /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(4371..4390)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4867..4887
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4997..5045
                     /note="synthetic polyadenylation signal"
     CDS             join(5311..5379,5380..6171)
                     /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(5529..5548)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6184..6320
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6472..6543
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6548..6594)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6612..6660
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatggcaat tgcctcagaa
      601 tttagttccc tcccctcgta cgctgcgttc gctaccgccc aagaggcata tgagcaggcc
      661 gtagccaacg gggacagcga agtggttctt aagaaactga agaaatcttt gaatgtggca
      721 aagagcgagt ttgatcggga cgcggcaatg cagaggaagc tggaaaaaat ggctgatcag
      781 gctatgacac agatgtacaa acaagcccga tccgaggaca agagagctaa agtcacttct
      841 gccatgcaaa cgatgctatt caccatgtta cgtaagctgg ataacgacgc actcaataac
      901 ataatcaata acgcccgcga tggttgtgtg ccattgaata tcattcctct tactacagcg
      961 gccaagctga tggtcgttat cccggactat aacacataca aaaatacctg cgacggcacc
     1021 actttcacgt atgctagcgc cctgtgggaa atacagcagg tagtggatgc cgattcaaag
     1081 attgtgcagc ttagtgagat ctccatggac aacagtccca acttggcttg gcctttaatc
     1141 gtcacagctc tcagagccaa tagcgcagtt aaactgcaag aaaaccttta ctttcagtcg
     1201 tacccctatg acgtaccaga ttatgcgtaa gctagcttga ctgactgaga tacagcgtac
     1261 cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1321 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1381 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     1441 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca
     1501 tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     1561 tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc
     1621 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     1681 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta
     1741 tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac
     1801 cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct
     1861 tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg
     1921 ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc
     1981 cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg
     2041 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2101 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2161 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2221 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2281 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2341 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2401 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2461 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2521 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2581 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2641 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2701 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2761 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2821 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     2881 tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa
     2941 acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg
     3001 ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc
     3061 gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta
     3121 tccgagatcc gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg
     3181 cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca cgtcagacga agggcgcagg
     3241 agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg
     3301 ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg
     3361 cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt
     3421 ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg
     3481 tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct
     3541 gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg
     3601 gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg
     3661 agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga
     3721 gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc
     3781 atggtgggcg gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa agctcttatt
     3841 cgggtgagat gggctgggca ccatctgggg accctgacgt gaagtttgtc actgactgga
     3901 gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca
     3961 cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt
     4021 ataatgcagg gtggggccac ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga
     4081 cgcagggttc gggcctaggg taggctctcc tgaatcgaca ggcgccggac ctctggtgag
     4141 gggagggata agtgaggcgt cagtttcttt ggtcggtttt atgtacctat cttcttaagt
     4201 agctgaagct ccggttttga actatgcgct cggggttggc gagtgtgttt tgtgaagttt
     4261 tttaggcacc ttttgaaatg taatcatttg ggtcaatatg taattttcag tgttagactt
     4321 gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacaac atgaccgagt
     4381 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     4441 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     4501 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     4561 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     4621 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     4681 ccgcgcagca acagatggag ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     4741 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     4801 tgctccccgg agtggaggcc gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     4861 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     4921 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctag gctagtatgt
     4981 aagcctagtc ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt
     5041 gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg
     5101 acgatccata tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt
     5161 ccaatcgtca gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct
     5221 agatattgat agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat
     5281 caagagacag gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt
     5341 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5401 gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac
     5461 agcggtaaga tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt
     5521 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5581 cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat
     5641 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5701 actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg
     5761 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5821 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa
     5881 ctattaactg gcgaactact tactctagct tcccggcaac agttgataga ctggatggag
     5941 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6001 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6061 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6121 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta accgattcta
     6181 ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
     6241 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
     6301 taccagaaat ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat
     6361 ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa
     6421 tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga
     6481 ccattagctc cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct
     6541 ctcaattcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc
     6601 gttgcgttac acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt
     6661 atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat
     6721 agcccatata tggagttccg
//
