LOCUS       sequence-295441-        4848 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 4848)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4848)
  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-m4134002
FEATURES             Location/Qualifiers
     source          1..4848
                     /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..664
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    717..838
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(728..747)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     782..801
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      953..999
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1029..1165
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1210..1281
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1507..2095)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1587..1606)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2841..2909,2910..3701)
                     /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(3059..3078)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3714..3850
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4002..4073
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4078..4124)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4142..4190
                     /note="synthetic polyadenylation signal"
     enhancer        4271..4574
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4576..4778
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4729..4749
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4847..4848,1..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 cggggaaacc tatacccaat ccacttctcg gcttagattc
      661 tacctaagct agcttgactg actgagatac agcgtacctt cagctcacag acatgataag
      721 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
      781 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
      841 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
      901 aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat cgtagcataa
      961 ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg ccggattgct
     1021 atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
     1081 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
     1141 ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt
     1201 cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag aagtagttca
     1261 tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa cttaattaac
     1321 cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg ctcagaattg
     1381 cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta cgagtttcat
     1441 ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1501 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1561 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1621 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1681 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1741 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1801 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1861 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1921 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1981 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2041 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2101 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2161 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2221 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2281 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg catttaaatt
     2341 tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc
     2401 aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc ttggctattg
     2461 cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat cgggatcacc
     2521 cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag gaatatcgaa
     2581 atcggggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata
     2641 tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca
     2701 gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat
     2761 agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag
     2821 gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt
     2881 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2941 ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3001 tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc
     3061 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3121 actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg
     3181 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3241 acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3301 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3361 acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg
     3421 gcgaactact tactctagct tcccggcaac agttgataga ctggatggag gcggataaag
     3481 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3541 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3601 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3661 agatcgctga gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg
     3721 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     3781 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     3841 ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt
     3901 tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc
     3961 tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc
     4021 cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa
     4081 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac
     4141 acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc
     4201 tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata
     4261 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     4321 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     4381 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     4441 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     4501 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     4561 tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     4621 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     4681 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     4741 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca
     4801 gatctttgtc gatcctacca tccactcgac acacccgcca gcggccgc
//
