LOCUS       sequence-314730-        7179 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 7179)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7179)
  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-m4133861
FEATURES             Location/Qualifiers
     source          1..7179
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1174..1194
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1195..1221
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1290..1411
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1301..1320)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1355..1374
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1526..1572
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1602..1738
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1783..1854
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2080..2668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2160..2179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3168..4377
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4214..4233
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4381..5409
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5436..5484
                     /note="synthetic polyadenylation signal"
     CDS             join(5750..5818,5819..6610)
                     /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(5968..5987)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6623..6759
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6911..6982
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6987..7033)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7051..7099
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgatagc ctcagagttt
      601 agttcccttc catcatatgc agcttttgct actgctcaag aagcttatga gcaggctgtt
      661 gctaatggtg attctgaagt tgttcttaaa aagttgaaga agtctttgaa tgtggctaaa
      721 tctgaatttg accgtgatgc agccatgcaa cgtaagttgg aaaagatggc tgatcaagct
      781 atgacccaaa tgtataaaca ggctagatct gaggacaaga gggcaaaagt tactagtgct
      841 atgcagacaa tgcttttcac tatgcttaga aagttggata atgatgcact caacaacatt
      901 atcaacaatg caagagatgg ttgtgttccc ttgaacataa tacctcttac aacagcagcc
      961 aaactaatgg ttgtcatacc agactataac acatataaaa atacgtgtga tggtacaaca
     1021 tttacttatg catcagcatt gtgggaaatc caacaggttg tagatgcaga tagtaaaatt
     1081 gttcaactta gtgaaattag tatggacaat tcacctaatt tagcatggcc tcttattgta
     1141 acagctttaa gggccaattc tgctgtcaaa ttagaaaacc tttactttca gtcgtacccc
     1201 tatgacgtac cagattatgc gtaaggatcc gcaggcctct gctagcttga ctgactgaga
     1261 tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac
     1321 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     1381 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     1441 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     1501 tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg
     1561 aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa
     1621 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     1681 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg
     1741 tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt
     1801 ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca
     1861 ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc
     1921 gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg
     1981 ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca
     2041 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2101 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2161 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2221 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2281 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2341 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2401 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2461 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2521 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2581 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2641 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2701 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2761 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2821 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2881 tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga
     2941 aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca
     3001 agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta
     3061 ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa
     3121 tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg
     3181 ggttttggcg cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca cgtcagacga
     3241 agggcgcagg agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca
     3301 taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt
     3361 gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt
     3421 ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac
     3481 gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt
     3541 gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg
     3601 tggccgccgg gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc
     3661 tgggtccgcg agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg
     3721 ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa
     3781 ggtggggggc atggtgggcg gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa
     3841 agctcttatt cgggtgagat gggctgggca ccatctgggg accctgacgt gaagtttgtc
     3901 actgactgga gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt
     3961 gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt
     4021 ctgttggctt ataatgcagg gtggggccac ctgccggtag gtgtgcggta ggcttttctc
     4081 cgtcgcagga cgcagggttc gggcctaggg taggctctcc tgaatcgaca ggcgccggac
     4141 ctctggtgag gggagggata agtgaggcgt cagtttcttt ggtcggtttt atgtacctat
     4201 cttcttaagt agctgaagct ccggttttga actatgcgct cggggttggc gagtgtgttt
     4261 tgtgaagttt tttaggcacc ttttgaaatg taatcatttg ggtcaatatg taattttcag
     4321 tgttagactt gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacaac
     4381 atgggtaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc
     4441 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc
     4501 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa
     4561 gatcgttatg tttatcggca ctttgcatcc gccgcgctcc cgattccgga agtgcttgac
     4621 attggggagt tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg
     4681 ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggcaatg
     4741 gatgccatcg ctgccgccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa
     4801 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg
     4861 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat
     4921 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc
     4981 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag
     5041 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg
     5101 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg
     5161 ccccggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt
     5221 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc
     5281 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgccgccgt ctggaccgat
     5341 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca
     5401 aaggaataag ctagtatgta agcctagtct tagataataa aatcgctatc catcgaagat
     5461 ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat
     5521 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     5581 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     5641 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     5701 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca
     5761 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5821 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta
     5881 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt
     5941 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     6001 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc
     6061 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6121 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa
     6181 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6241 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6301 ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6361 gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6421 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     6481 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     6541 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6601 gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     6661 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     6721 cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc gtttaaactc
     6781 gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat
     6841 ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg
     6901 cggctcccga catcttggac cattagctcc acaggtatct tcttccctct agtggtcata
     6961 acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca
     7021 aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc
     7081 gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat
     7141 tacggggtca ttagttcata gcccatatat ggagttccg
//
