LOCUS       sequence-357052-        6521 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF8 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6521)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6521)
  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-m4133899
FEATURES             Location/Qualifiers
     source          1..6521
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             368..388
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             389..430
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    483..604
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(494..513)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     548..567
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      719..765
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      795..931
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    976..1047
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1273..1861)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1353..1372)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2361..3570
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3407..3426
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3574..4173
                     /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(3574..3593)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4070..4090
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4200..4248
                     /note="synthetic polyadenylation signal"
     CDS             join(4514..4582,4583..5374)
                     /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(4732..4751)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5387..5523
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5675..5746
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5751..5797)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5815..5863
                     /note="synthetic polyadenylation signal"
     enhancer        5944..6247
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6249..6451
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6402..6422
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaaa ttcttagtgt ttcttgggat cattacaacg gtcgcagctt ttcatcagga
       61 atgcagccta caatcgtgta cccagcacca gccgtacgtt gtggacgatc cttgccccat
      121 ccacttctat tctaagtggt acatacgggt aggcgcgagg aagagtgcac cactcattga
      181 gttgtgtgtg gacgaggccg gttcaaaatc cccaatccag tatatagata ttggaaatta
      241 cactgtctcc tgtctgccct tcaccatcaa ctgccaagaa cctaagctgg gaagcctcgt
      301 tgtgagatgc tctttttatg aggattttct tgaataccac gacgtccgtg tggttctgga
      361 cttcattgaa aacctttact ttcagtcggg gaaacctata cccaatccac ttctcggctt
      421 agattctacc taagctagct tgactgactg agatacagcg taccttcagc tcacagacat
      481 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
      541 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
      601 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
      661 tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta
      721 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg
      781 attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
      841 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
      901 cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct
      961 cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt
     1021 agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta
     1081 attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca
     1141 gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag
     1201 tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1261 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1321 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1381 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1441 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1501 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1561 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1621 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1681 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1741 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1801 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     1861 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     1921 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     1981 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2041 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt
     2101 taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca
     2161 tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct tgcgccttgg
     2221 ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc cgagatcggg
     2281 atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga tccgaggaat
     2341 atcgaaatcg gggcgcgcct ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc
     2401 cctcgtcacg gcgagcgctg ccacgtcaga cgaagggcgc aggagcgtcc tgatccttcc
     2461 gcccggacgc tcaggacagc ggcccgctgc tcataagact cggccttaga accccagtat
     2521 cagcagaagg acattttagg acgggacttg ggtgactcta gggcactggt tttctttcca
     2581 gagagcggaa caggcgagga aaagtagtcc cttctcggcg attctgcgga gggatctccg
     2641 tggggcggtg aacgccgatg attatataag gacgcgccgg gtgtggcaca gctagttccg
     2701 tcgcagccgg gatttgggtc gcggttcttg tttgtggatc gctgtgatcg tcacttggtg
     2761 agtagcgggc tgctgggctg gccggggctt tcgtggccgc cgggccgctc ggtgggacgg
     2821 aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc gcgagcaagg ttgccctgaa
     2881 ctgggggttg gggggagcgc agcaaaatgg cggctgttcc cgagtcttga atggaagacg
     2941 cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg ggcatggtgg gcggcaagaa
     3001 cccaaggtct tgagcccttc gctaatgcgg gaaagctctt attcgggtga gatgggctgg
     3061 gcaccatctg gggaccctga cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc
     3121 tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt gcacccgtac ctttgggagc
     3181 gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc agggtggggc
     3241 cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta
     3301 gggtaggctc tcctgaatcg acaggcgccg gacctctggt gaggggaggg ataagtgagg
     3361 cgtcagtttc tttggtcggt tttatgtacc tatcttctta agtagctgaa gctccggttt
     3421 tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa
     3481 atgtaatcat ttgggtcaat atgtaatttt cagtgttaga cttgtaaatt gtccgctaaa
     3541 ttctggccgt ttttggcttt tttgttagac aacatgaccg agtacaagcc cacggtgcgc
     3601 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     3661 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     3721 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     3781 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     3841 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     3901 gagggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     3961 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     4021 gccgccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     4081 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     4141 acctggtgca tgacccgcaa gcccggtgcc taggctagta tgtaagccta gtcttagata
     4201 ataaaatcgc tatccatcga agatggatgt gtgttggttt tttgtgtgtg taacgctagg
     4261 cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc
     4321 ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg tcagatattg
     4381 tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt gatagtctga
     4441 tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga caggatcagc
     4501 aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4561 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4621 tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4681 gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc tgctatgtgg
     4741 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4801 tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg atggcatgac
     4861 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4921 tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4981 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5041 tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa ctggcgaact
     5101 acttactcta gcttcccggc aacagttgat agactggatg gaggcggata aagttgcagg
     5161 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     5221 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     5281 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     5341 tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa
     5401 aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg
     5461 gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct
     5521 taagatcccg aatcgtttaa actcgactct ggctctatcg aatctccgtc gtttcgagct
     5581 tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc
     5641 agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag ctccacaggt
     5701 atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt caaaaaaccc
     5761 ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt tacacacaca
     5821 aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac tgctgatcga
     5881 gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat atatggagtt
     5941 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     6001 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     6061 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     6121 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     6181 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     6241 taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     6301 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     6361 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     6421 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tcagatcttt
     6481 gtcgatccta ccatccactc gacacacccg ccagcggccg c
//
