LOCUS       sequence-295115-        5355 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 5355)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5355)
  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-m4133137
FEATURES             Location/Qualifiers
     source          1..5355
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             596..616
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             650..727
                     /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             755..775
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1224..1345
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1235..1254)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1289..1308
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1460..1506
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1536..1672
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1717..1788
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2014..2602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2094..2113)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3348..3416,3417..4208)
                     /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(3566..3585)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4221..4357
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4509..4580
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4585..4631)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4649..4697
                     /note="synthetic polyadenylation signal"
     enhancer        4778..5081
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5083..5285
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5236..5256
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgata gcctcagagt ttagttccct tccatcatat gcagcttttg ctactgctca
       61 agaagcttat gagcaggctg ttgctaatgg tgattctgaa gttgttctta aaaagttgaa
      121 gaagtctttg aatgtggcta aatctgaatt tgaccgtgat gcagccatgc aacgtaagtt
      181 ggaaaagatg gctgatcaag ctatgaccca aatgtataaa caggctagat ctgaggacaa
      241 gagggcaaaa gttactagtg ctatgcagac aatgcttttc actatgctta gaaagttgga
      301 taatgatgca ctcaacaaca ttatcaacaa tgcaagagat ggttgtgttc ccttgaacat
      361 aatacctctt acaacagcag ccaaactaat ggttgtcata ccagactata acacatataa
      421 aaatacgtgt gatggtacaa catttactta tgcatcagca ttgtgggaaa tccaacaggt
      481 tgtagatgca gatagtaaaa ttgttcaact tagtgaaatt agtatggaca attcacctaa
      541 tttagcatgg cctcttattg taacagcttt aagggccaat tctgctgtca aattagaaaa
      601 cctttacttt cagtcgggac ggcgaatccc ggggctgata aatccctgga aaaggagatg
      661 gaagaagaac tttattgcag tgtctgcggc taaccggttc aaaaagatca gttcaagcgg
      721 cgccctagat tacgacattc ctacgaccgc ctccgagaat ctctatttcc agggtgaatt
      781 tggcttggca caacatgacg aggccgtcga taacaagttt aataaagaac agcagaacgc
      841 tttctatgag atccttcact taccaaatct gaacgaagag caacgcaacg ctttcattca
      901 gagcctgaaa gacgatccct cgcagtctgc aaatctcctg gccgaagcca agaagctgaa
      961 tgacgcacag gctccaaaag ttgataacaa gtttaacaag gagcagcaga atgctttcta
     1021 cgagatattg caccttccta atctgaatga ggaacaaaga aatgcattta tccagtccct
     1081 aaaagacgat ccatcacaga gtgccaacct gcttgcggaa gccaaaaagc tcaatgatgc
     1141 ccaggccccg aaagtagacg cgaaccatca ataagctagc ttgactgact gagatacagc
     1201 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1261 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1321 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1381 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg
     1441 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1501 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg
     1561 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     1621 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca
     1681 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg
     1741 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt
     1801 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg
     1861 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg
     1921 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc
     1981 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2041 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2101 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2161 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2221 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2281 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2341 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2401 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2461 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2521 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2581 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2641 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2701 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2761 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2821 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct
     2881 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc
     2941 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa
     3001 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga
     3061 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggtgtaccg agaacgatcc
     3121 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag
     3181 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta
     3241 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag
     3301 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac
     3361 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3421 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca
     3481 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc
     3541 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3601 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac
     3661 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3721 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg
     3781 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3841 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3901 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt
     3961 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4021 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4081 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4141 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4201 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     4261 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     4321 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga
     4381 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt
     4441 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg
     4501 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac
     4561 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag
     4621 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga
     4681 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta
     4741 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     4801 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     4861 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     4921 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     4981 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     5041 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt
     5101 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     5161 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     5221 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     5281 gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca
     5341 cccgccagcg gccgc
//
