LOCUS       sequence-297964-        4869 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 4869)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4869)
  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-m4133178
FEATURES             Location/Qualifiers
     source          1..4869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        40..343
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        345..547
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     498..518
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1213..1233
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1237..1302
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1355..1476
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1366..1385)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1420..1439
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1591..1637
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1667..1803
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1848..1919
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2145..2733)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2225..2244)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3479..3547,3548..4339)
                     /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(3697..3716)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4352..4488
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4640..4711
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4716..4762)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4780..4828
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
       61 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      121 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      181 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      241 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      301 tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca
      361 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      421 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      481 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      541 cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca
      601 cacccgccag cggccgccac catgatagcc tcagagttta gttcccttcc atcatatgca
      661 gcttttgcta ctgctcaaga agcttatgag caggctgttg ctaatggtga ttctgaagtt
      721 gttcttaaaa agttgaagaa gtctttgaat gtggctaaat ctgaatttga ccgtgatgca
      781 gccatgcaac gtaagttgga aaagatggct gatcaagcta tgacccaaat gtataaacag
      841 gctagatctg aggacaagag ggcaaaagtt actagtgcta tgcagacaat gcttttcact
      901 atgcttagaa agttggataa tgatgcactc aacaacatta tcaacaatgc aagagatggt
      961 tgtgttccct tgaacataat acctcttaca acagcagcca aactaatggt tgtcatacca
     1021 gactataaca catataaaaa tacgtgtgat ggtacaacat ttacttatgc atcagcattg
     1081 tgggaaatcc aacaggttgt agatgcagat agtaaaattg ttcaacttag tgaaattagt
     1141 atggacaatt cacctaattt agcatggcct cttattgtaa cagctttaag ggccaattct
     1201 gctgtcaaat tagaaaacct ttactttcag tcgatggatt ataaggacca cgatggagac
     1261 tacaaagatc atgacattga ttacaaggac gacgatgaca aataagctag cttgactgac
     1321 tgagatacag cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa
     1381 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     1441 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     1501 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     1561 tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg
     1621 tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa
     1681 aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg
     1741 gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct
     1801 taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg
     1861 gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa
     1921 tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg
     1981 agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt
     2041 ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc
     2101 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2161 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2221 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2281 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2341 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2401 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2461 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2521 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2581 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2641 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2701 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2761 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2821 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2881 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2941 atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg
     3001 tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta
     3061 tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg
     3121 tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc
     3181 ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc ctggtgtacc
     3241 gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag
     3301 tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc
     3361 acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa
     3421 tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat
     3481 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3541 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg
     3601 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3661 agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3721 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3781 tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3841 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg
     3901 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3961 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4021 tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc
     4081 ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4141 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4201 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4261 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4321 actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc
     4381 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     4441 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga tcccgaatcg
     4501 tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt
     4561 ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg
     4621 cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta
     4681 gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta
     4741 gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca
     4801 tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta
     4861 gtaatcaat
//
