LOCUS       sequence-295363-        5196 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 5196)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5196)
  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-m4134453
FEATURES             Location/Qualifiers
     source          1..5196
                     /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             971..988
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1065..1186
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1076..1095)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1130..1149
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1301..1347
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1377..1513
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1558..1629
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1855..2443)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1935..1954)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3189..3257,3258..4049)
                     /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(3407..3426)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4062..4198
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4350..4421
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4426..4472)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4490..4538
                     /note="synthetic polyadenylation signal"
     enhancer        4619..4922
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4924..5126
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5077..5097
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5195..5196,1..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 cgatgagtga caaaatcatt catctaaccg atgactcctt
      661 tgatacagac gtgttgaagg ccgatggtgc gatactggta gatttctggg ctgaatggtg
      721 tggcccgtgc aagatgatcg cacccattct ggacgagatc gccgacgagt atcaaggaaa
      781 attaactgtc gctaagctta atatagatca gaacccaggg acggcaccta aatacggcat
      841 caggggaata cctaccctcc tgctctttaa gaatggggaa gttgccgcga caaaagtggg
      901 agccctgtct aagggccagc taaaagagtt cttggacgct aacctggcag gttcaggctc
      961 ggggcacatg caccatcacc atcaccatag cagcgggctc gtgccacgcg gctaagctag
     1021 cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat acattgatga
     1081 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     1141 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1201 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     1261 cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc
     1321 tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat
     1381 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     1441 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     1501 aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat
     1561 tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt
     1621 ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag
     1681 ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt
     1741 tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg
     1801 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1861 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1921 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1981 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2041 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2101 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2161 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2221 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2281 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2341 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2401 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2461 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2521 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2581 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2641 tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc
     2701 aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc
     2761 gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc
     2821 tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc
     2881 aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc
     2941 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     3001 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     3061 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     3121 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     3181 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3241 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3301 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3361 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3421 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3481 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3541 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3601 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3661 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3721 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     3781 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     3841 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3901 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3961 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4021 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     4081 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     4141 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     4201 tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc
     4261 gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt
     4321 acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt
     4381 cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa
     4441 gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa
     4501 ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta
     4561 gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     4621 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     4681 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     4741 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     4801 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     4861 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     4921 tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     4981 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     5041 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     5101 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga
     5161 tcctaccatc cactcgacac acccgccagc ggccgc
//
