LOCUS       sequence-295357-        5361 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 5361)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5361)
  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-m4134437
FEATURES             Location/Qualifiers
     source          1..5361
                     /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             656..733
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             761..781
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1230..1351
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1241..1260)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1295..1314
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1466..1512
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1542..1678
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1723..1794
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2020..2608)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2100..2119)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3354..3422,3423..4214)
                     /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(3572..3591)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4227..4363
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4515..4586
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4591..4637)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4655..4703
                     /note="synthetic polyadenylation signal"
     enhancer        4784..5087
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5089..5291
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5242..5262
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5360..5361,1..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 cgggacggcg aatcccgggg ctgataaatc cctggaaaag
      661 gagatggaag aagaacttta ttgcagtgtc tgcggctaac cggttcaaaa agatcagttc
      721 aagcggcgcc ctagattacg acattcctac gaccgcctcc gagaatctct atttccaggg
      781 tgaatttggc ttggcacaac atgacgaggc cgtcgataac aagtttaata aagaacagca
      841 gaacgctttc tatgagatcc ttcacttacc aaatctgaac gaagagcaac gcaacgcttt
      901 cattcagagc ctgaaagacg atccctcgca gtctgcaaat ctcctggccg aagccaagaa
      961 gctgaatgac gcacaggctc caaaagttga taacaagttt aacaaggagc agcagaatgc
     1021 tttctacgag atattgcacc ttcctaatct gaatgaggaa caaagaaatg catttatcca
     1081 gtccctaaaa gacgatccat cacagagtgc caacctgctt gcggaagcca aaaagctcaa
     1141 tgatgcccag gccccgaaag tagacgcgaa ccatcaataa gctagcttga ctgactgaga
     1201 tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac
     1261 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     1321 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     1381 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     1441 tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg
     1501 aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa
     1561 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     1621 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg
     1681 tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt
     1741 ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca
     1801 ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc
     1861 gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg
     1921 ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca
     1981 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2041 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2101 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2161 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2221 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2281 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2341 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2401 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2461 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2521 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2581 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2641 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2701 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2761 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2821 tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga
     2881 aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca
     2941 agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta
     3001 ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa
     3061 tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa
     3121 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     3181 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     3241 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     3301 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     3361 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3421 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     3481 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3541 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3601 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3661 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3721 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     3781 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3841 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3901 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     3961 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4021 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4081 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4141 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4201 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     4261 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     4321 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa
     4381 actcgactct ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc
     4441 tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg
     4501 atcgcggctc ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt
     4561 cataacagca gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc
     4621 cccaaggggt tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat
     4681 cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat
     4741 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     4801 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     4861 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     4921 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     4981 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     5041 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt
     5101 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     5161 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     5221 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     5281 taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc
     5341 gacacacccg ccagcggccg c
//
