LOCUS       sequence-295380-        6944 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 6944)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6944)
  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-m4133612
FEATURES             Location/Qualifiers
     source          1..6944
                     /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..415
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    484..605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(495..514)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     549..568
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      720..766
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      796..932
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    977..1048
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1274..1862)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1354..1373)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2362..3571
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3408..3427
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3575..4603
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4630..4678
                     /note="synthetic polyadenylation signal"
     CDS             join(4944..5012,5013..5804)
                     /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(5162..5181)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5817..5953
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6098..6169
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6174..6220)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6238..6286
                     /note="synthetic polyadenylation signal"
     enhancer        6367..6670
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6672..6874
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6825..6845
                     /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 ttcagtcgta cccctatgac gtaccagatt atgcgtaagg
      421 atccgcaggc ctctgctagc ttgactgact gagatacagc gtaccttcag ctcacagaca
      481 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
      541 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
      601 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
      661 ttttttaaag caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt
      721 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg
      781 gattgctatc taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta
      841 tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat
      901 acgtgtcctc cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc
      961 tcgatttcga tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag
     1021 tagttcatca aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt
     1081 aattaaccag tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc
     1141 agaattgcgt cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga
     1201 gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1261 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1321 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1381 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1441 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1501 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1561 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1621 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1681 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     1741 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     1801 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1861 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     1921 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     1981 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2041 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat
     2101 ttaaatttcc gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc
     2161 atcttgcagg ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg
     2221 gctattgctt ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg
     2281 gatcacccga gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa
     2341 tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc
     2401 ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg caggagcgtc ctgatccttc
     2461 cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta
     2521 tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc
     2581 agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc
     2641 gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc
     2701 gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt
     2761 gagtagcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg
     2821 gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga
     2881 actgggggtt ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac
     2941 gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga
     3001 acccaaggtc ttgagccctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg
     3061 ggcaccatct ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt
     3121 ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag
     3181 cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg
     3241 ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct
     3301 agggtaggct ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag
     3361 gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt
     3421 ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga
     3481 aatgtaatca tttgggtcaa tatgtaattt tcagtgttag acttgtaaat tgtccgctaa
     3541 attctggccg tttttggctt ttttgttaga caacatgggt aaaaagcctg aactcaccgc
     3601 gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct
     3661 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct
     3721 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc
     3781 atccgccgcg ctcccgattc cggaagtgct tgacattggg gagttcagcg agagcctgac
     3841 ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact
     3901 gcccgctgtt ctgcagccgg tcgcggaggc aatggatgcc atcgctgccg ccgatcttag
     3961 ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg
     4021 tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga
     4081 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg
     4141 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa
     4201 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga
     4261 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta
     4321 cttcgagcgg aggcatccgg agcttgcagg atcgccccgg ctccgggcgt atatgctccg
     4381 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg
     4441 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca
     4501 aatcgcccgc agaagcgccg ccgtctggac cgatggctgt gtagaagtac tcgccgatag
     4561 tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa taagctagta tgtaagccta
     4621 gtcttagata ataaaatcgc tatccatcga agatggatgt gtgttggttt tttgtgtgtg
     4681 taacgctagg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc
     4741 atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg
     4801 tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt
     4861 gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga
     4921 caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct
     4981 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5041 atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta
     5101 agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc
     5161 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5221 tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg
     5281 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5341 ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca
     5401 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5461 acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa
     5521 ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg gaggcggata
     5581 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5641 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5701 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5761 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat
     5821 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     5881 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     5941 aatttatcct taagatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga
     6001 gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc
     6061 gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat tagctccaca
     6121 ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca attcaaaaaa
     6181 cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg cgttacacac
     6241 acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat
     6301 cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc catatatgga
     6361 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     6421 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     6481 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     6541 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     6601 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     6661 tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     6721 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     6781 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     6841 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatcagatc
     6901 tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgc
//
