LOCUS       sequence-295009-        4818 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 4818)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4818)
  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-m4133696
FEATURES             Location/Qualifiers
     source          1..4818
                     /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..634
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    687..808
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(698..717)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     752..771
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      923..969
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      999..1135
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1180..1251
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1477..2065)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1557..1576)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2811..2879,2880..3671)
                     /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(3029..3048)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3684..3820
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3972..4043
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4048..4094)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4112..4160
                     /note="synthetic polyadenylation signal"
     enhancer        4241..4544
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4546..4748
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4699..4719
                     /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 cagtcgcacc atcaccatca ccattaagct agcttgactg actgagatac
      661 agcgtacctt cagctcacag acatgataag atacattgat gagtttggac aaaccacaac
      721 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
      781 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
      841 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
      901 tggcccatct ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg
      961 ggttttttgt gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc
     1021 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     1081 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg
     1141 tcagctatcc tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc
     1201 tggacaccac taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg
     1261 gttaccttgg gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac
     1321 ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta
     1381 ttgcacccgt tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa
     1441 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1501 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1561 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1621 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1681 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1741 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1801 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1861 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1921 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1981 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2041 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2101 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2161 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2221 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2281 atttcgttca tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa
     2341 tcttcaaacc tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt
     2401 ctcttggccg gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc
     2461 caatcccgaa tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc
     2521 cgatctatcc gagatccgag gaatatcgaa atcggggcgc gcctggtgta ccgagaacga
     2581 tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc
     2641 cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc
     2701 gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc
     2761 tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc
     2821 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2881 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt
     2941 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct
     3001 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3061 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt
     3121 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3181 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga
     3241 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3301 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3361 tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3421 agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3481 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3541 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3601 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3661 agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc
     3721 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
     3781 acttccatac gtgtcctcct taccagaaat ttatccttaa gatcccgaat cgtttaaact
     3841 cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca
     3901 tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc
     3961 gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc tagtggtcat
     4021 aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt tagaggcccc
     4081 aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca catccatctt
     4141 cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc tagtaatcaa
     4201 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     4261 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     4321 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     4381 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     4441 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     4501 ctacttggca gtacatctac gtattagtca tcgctattac catgctgatg cggttttggc
     4561 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     4621 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     4681 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     4741 gcagagctgg tttagtgaac cgtcagatca gatctttgtc gatcctacca tccactcgac
     4801 acacccgcca gcggccgc
//
