LOCUS       sequence-295447-        4845 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 4845)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4845)
  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-m4133147
FEATURES             Location/Qualifiers
     source          1..4845
                     /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             617..661
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    714..835
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(725..744)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     779..798
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      950..996
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1026..1162
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1207..1278
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1504..2092)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1584..1603)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2838..2906,2907..3698)
                     /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(3056..3075)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3711..3847
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3999..4070
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4075..4121)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4139..4187
                     /note="synthetic polyadenylation signal"
     enhancer        4268..4571
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4573..4775
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4726..4746
                     /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 cagtcgaaag aaaccgcggc agctaagttc gagaggcagc acatggacag
      661 ttaagctagc ttgactgact gagatacagc gtaccttcag ctcacagaca tgataagata
      721 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
      781 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
      841 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
      901 caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt agcataaccc
      961 cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg gattgctatc
     1021 taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     1081 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     1141 cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc tcgatttcga
     1201 tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag tagttcatca
     1261 aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt aattaaccag
     1321 tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt
     1381 cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta
     1441 aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1501 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1561 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1621 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1681 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1741 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1801 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1861 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1921 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     1981 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2041 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2101 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2161 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2221 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2281 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc
     2341 gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg
     2401 ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt
     2461 ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga
     2521 gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc
     2581 ggggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg
     2641 ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat
     2701 attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt
     2761 ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat
     2821 cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2881 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2941 aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3001 ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat
     3061 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3121 attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca
     3181 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3241 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3301 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3361 agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg
     3421 aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg
     3481 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3541 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3601 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3661 tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc
     3721 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     3781 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     3841 tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg
     3901 agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat
     3961 cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac
     4021 aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa
     4081 acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca
     4141 cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc taactgctga
     4201 tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg
     4261 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     4321 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     4381 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     4441 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     4501 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     4561 ctattaccat gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     4621 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     4681 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     4741 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat
     4801 ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgc
//
