LOCUS       sequence-300228-        6734 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 6734)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6734)
  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-m4133908
FEATURES             Location/Qualifiers
     source          1..6734
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        114..417
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        419..621
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     572..592
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1287..1307
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1308..1334
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1387..1508
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1398..1417)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     terminator      1623..1669
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1699..1835
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1880..1951
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2177..2765)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2257..2276)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3265..4474
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4311..4330
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4478..5077
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4478..4497)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4974..4994
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5104..5152
                     /note="synthetic polyadenylation signal"
     CDS             join(5418..5486,5487..6278)
                     /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(5636..5655)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6291..6427
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6579..6650
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6655..6701)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    join(6719..6734,1..33)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 acacatccat cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag
       61 atctagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      121 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      181 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      241 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      301 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      361 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg
      421 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      481 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      541 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      601 gaggtctata taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta
      661 ccatccactc gacacacccg ccagcggccg ccaccatgat agcctcagag tttagttccc
      721 ttccatcata tgcagctttt gctactgctc aagaagctta tgagcaggct gttgctaatg
      781 gtgattctga agttgttctt aaaaagttga agaagtcttt gaatgtggct aaatctgaat
      841 ttgaccgtga tgcagccatg caacgtaagt tggaaaagat ggctgatcaa gctatgaccc
      901 aaatgtataa acaggctaga tctgaggaca agagggcaaa agttactagt gctatgcaga
      961 caatgctttt cactatgctt agaaagttgg ataatgatgc actcaacaac attatcaaca
     1021 atgcaagaga tggttgtgtt cccttgaaca taatacctct tacaacagca gccaaactaa
     1081 tggttgtcat accagactat aacacatata aaaatacgtg tgatggtaca acatttactt
     1141 atgcatcagc attgtgggaa atccaacagg ttgtagatgc agatagtaaa attgttcaac
     1201 ttagtgaaat tagtatggac aattcaccta atttagcatg gcctcttatt gtaacagctt
     1261 taagggccaa ttctgctgtc aaattagaaa acctttactt tcagtcgtac ccctatgacg
     1321 taccagatta tgcgtaagct agcttgactg actgagatac agcgtacctt cagctcacag
     1381 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     1441 gctttatttg tgaaatttgt gatgctattg atttatttgt aaccattata agctgcaata
     1501 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     1561 aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat
     1621 cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg
     1681 ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc
     1741 ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc
     1801 catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat
     1861 ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag
     1921 aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa
     1981 cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg
     2041 ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta
     2101 cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2161 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2221 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2281 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2341 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2401 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2461 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2521 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2581 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2641 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2701 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2761 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2821 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2881 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     2941 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3001 catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga
     3061 tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc
     3121 ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat
     3181 cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag
     3241 gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg
     3301 cccccctcgt cacggcgagc gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc
     3361 ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct tagaacccca
     3421 gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac tggttttctt
     3481 tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg cggagggatc
     3541 tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt
     3601 tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt
     3661 ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc gctcggtggg
     3721 acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc
     3781 tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa
     3841 gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca
     3901 agaacccaag gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg gtgagatggg
     3961 ctgggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt
     4021 cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg
     4081 gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg
     4141 gggccacctg ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg
     4201 cctagggtag gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt
     4261 gaggcgtcag tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg
     4321 gttttgaact atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt
     4381 tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagacttgta aattgtccgc
     4441 taaattctgg ccgtttttgg cttttttgtt agacaacatg accgagtaca agcccacggt
     4501 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     4561 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     4621 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     4681 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     4741 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     4801 gatggagggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     4861 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     4921 ggaggccgcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     4981 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     5041 gcgcacctgg tgcatgaccc gcaagcccgg tgcctaggct agtatgtaag cctagtctta
     5101 gataataaaa tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc
     5161 taggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg
     5221 ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat
     5281 attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt
     5341 ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat
     5401 cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5461 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5521 aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5581 ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat
     5641 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5701 attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca
     5761 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5821 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5881 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5941 agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg
     6001 aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg
     6061 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6121 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6181 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6241 tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc
     6301 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     6361 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     6421 tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg
     6481 agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat
     6541 cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac
     6601 aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa
     6661 acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca
     6721 cacaaaaaac caac
//
