LOCUS       sequence-295836-        6959 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 6959)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6959)
  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-m4133553
FEATURES             Location/Qualifiers
     source          1..6959
                     /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    499..620
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(510..529)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     564..583
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      735..781
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      811..947
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    992..1063
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1289..1877)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1369..1388)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2377..3586
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3423..3442
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3590..4618
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4645..4693
                     /note="synthetic polyadenylation signal"
     CDS             join(4959..5027,5028..5819)
                     /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(5177..5196)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5832..5968
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6113..6184
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6189..6235)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6253..6301
                     /note="synthetic polyadenylation signal"
     enhancer        6382..6685
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6687..6889
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6840..6860
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaaa tttcttgttt tcttaggaat catcacaact gtagctgcat ttcaccaaga
       61 atgtagttta cagtcatgta ctcaacatca accatatgta gttgatgacc cgtgtcctat
      121 tcacttctat tctaaatggt atattagagt aggagctaga aaatcagcac ctttaattga
      181 attgtgcgtg gatgaggctg gttctaaatc acccattcag tacatcgata tcggtaatta
      241 tacagtttcc tgtttacctt ttacaattaa ttgccaggaa cctaaattgg gtagtcttgt
      301 agtgcgttgt tcgttctatg aagacttttt agagtatcat gacgttcgtg ttgttttaga
      361 tttcatcgaa aacctttact ttcagtcggg gaaacctata cccaatccac ttctcggctt
      421 agattctacc taaggatccg caggcctctg ctagcttgac tgactgagat acagcgtacc
      481 ttcagctcac agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
      541 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
      601 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
      661 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat
      721 ctctatcggt atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
      781 gtgcccctcg ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg
      841 acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa
      901 cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat
      961 cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc
     1021 actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt
     1081 gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg
     1141 gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc
     1201 gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga
     1261 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1321 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1381 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1441 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1501 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1561 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1621 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1681 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1741 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1801 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1861 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1921 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1981 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2041 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2101 catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa
     2161 cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc
     2221 cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg
     2281 aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat
     2341 ccgagatccg aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc
     2401 ctcccgcggg cgcccccctc gtcacggcga gcgctgccac gtcagacgaa gggcgcagga
     2461 gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc
     2521 cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc
     2581 actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc
     2641 tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt
     2701 ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg
     2761 tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg
     2821 ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga
     2881 gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag
     2941 tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca
     3001 tggtgggcgg caagaaccca aggtcttgag cccttcgcta atgcgggaaa gctcttattc
     3061 gggtgagatg ggctgggcac catctgggga ccctgacgtg aagtttgtca ctgactggag
     3121 aactcggttt gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac
     3181 ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta
     3241 taatgcaggg tggggccacc tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac
     3301 gcagggttcg ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg
     3361 ggagggataa gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta
     3421 gctgaagctc cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt
     3481 ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagacttg
     3541 taaattgtcc gctaaattct ggccgttttt ggcttttttg ttagacaaca tgggtaaaaa
     3601 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     3661 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     3721 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     3781 ttatcggcac tttgcatccg ccgcgctccc gattccggaa gtgcttgaca ttggggagtt
     3841 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     3901 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggcaatgg atgccatcgc
     3961 tgccgccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     4021 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     4081 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     4141 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     4201 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     4261 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     4321 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cccggctccg
     4381 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     4441 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     4501 tgtcgggcgt acacaaatcg cccgcagaag cgccgccgtc tggaccgatg gctgtgtaga
     4561 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaataagc
     4621 tagtatgtaa gcctagtctt agataataaa atcgctatcc atcgaagatg gatgtgtgtt
     4681 ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc gagaacgatc ctctcagtgc
     4741 gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca gcttgggacc
     4801 caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt accgatctgt
     4861 ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta gcttttgcaa
     4921 acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa catttccgtg
     4981 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5041 tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac atcgaactgg
     5101 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt ccaatgatga
     5161 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5221 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca ccagtcacag
     5281 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5341 gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag gagctaaccg
     5401 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5461 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct
     5521 tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag ttgatagact
     5581 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5641 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5701 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5761 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5821 cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     5881 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     5941 gtcctcctta ccagaaattt atccttaaga tcgtttaaac tcgactctgg ctctatcgaa
     6001 tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga
     6061 atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg
     6121 accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc
     6181 tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat
     6241 cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat
     6301 tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca
     6361 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     6421 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     6481 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     6541 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     6601 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     6661 cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca atgggcgtgg
     6721 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     6781 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     6841 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     6901 ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgc
//
