LOCUS       sequence-295257-        4824 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 4824)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4824)
  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-m4134147
FEATURES             Location/Qualifiers
     source          1..4824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             602..622
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             623..640
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    693..814
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(704..723)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     758..777
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      929..975
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1005..1141
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1186..1257
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1483..2071)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1563..1582)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2817..2885,2886..3677)
                     /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(3035..3054)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3690..3826
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3978..4049
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4054..4100)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4118..4166
                     /note="synthetic polyadenylation signal"
     enhancer        4247..4550
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4552..4754
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4705..4725
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4823..4824,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggca attgcctcag aatttagttc cctcccctcg tacgctgcgt tcgctaccgc
       61 ccaagaggca tatgagcagg ccgtagccaa cggggacagc gaagtggttc ttaagaaact
      121 gaagaaatct ttgaatgtgg caaagagcga gtttgatcgg gacgcggcaa tgcagaggaa
      181 gctggaaaaa atggctgatc aggctatgac acagatgtac aaacaagccc gatccgagga
      241 caagagagct aaagtcactt ctgccatgca aacgatgcta ttcaccatgt tacgtaagct
      301 ggataacgac gcactcaata acataatcaa taacgcccgc gatggttgtg tgccattgaa
      361 tatcattcct cttactacag cggccaagct gatggtcgtt atcccggact ataacacata
      421 caaaaatacc tgcgacggca ccactttcac gtatgctagc gccctgtggg aaatacagca
      481 ggtagtggat gccgattcaa agattgtgca gcttagtgag atctccatgg acaacagtcc
      541 caacttggct tggcctttaa tcgtcacagc tctcagagcc aatagcgcag ttaaactgca
      601 agaaaacctt tactttcagt cgcaccatca ccatcaccat taagctagct tgactgactg
      661 agatacagcg taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac
      721 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
      781 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
      841 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
      901 tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc
      961 ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa
     1021 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     1081 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     1141 aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt
     1201 cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc
     1261 tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag
     1321 atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc
     1381 ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca
     1441 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1501 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1561 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1621 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1681 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1741 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1801 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1861 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1921 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1981 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2041 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2101 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2161 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2221 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2281 ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc
     2341 ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc
     2401 gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta
     2461 ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct
     2521 caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggtgtaccga
     2581 gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc
     2641 ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac
     2701 ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc
     2761 gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga
     2821 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2881 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag
     2941 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3001 aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3061 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3121 agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3181 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgattggag
     3241 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3301 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3361 tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc
     3421 ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3481 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3541 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3601 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3661 tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga
     3721 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
     3781 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt
     3841 taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg
     3901 cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca
     3961 gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt
     4021 ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga
     4081 ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc
     4141 catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt
     4201 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     4261 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     4321 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     4381 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     4441 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     4501 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt
     4561 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     4621 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     4681 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     4741 atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca
     4801 ctcgacacac ccgccagcgg ccgc
//
