LOCUS       sequence-306086-        6951 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 6951)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6951)
  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..6951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    102..173
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(178..224)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    242..290
                     /note="synthetic polyadenylation signal"
     enhancer        371..674
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        676..878
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     829..849
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1316..1336
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1337..1363
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1432..1553
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1443..1462)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1497..1516
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1668..1714
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1744..1880
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1925..1996
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2222..2810)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2302..2321)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3310..4519
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4356..4375
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4523..5551
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5578..5626
                     /note="synthetic polyadenylation signal"
     CDS             join(5892..5960,5961..6752)
                     /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(6110..6129)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6765..6901
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc
       61 tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc
      121 cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa
      181 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac
      241 acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc
      301 tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata
      361 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      421 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      481 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      541 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      601 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      661 tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      721 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      781 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      841 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca
      901 gatctttgtc gatcctacca tccactcgac acacccgcca gcggccgcca ccatgaaatt
      961 cttagtgttt cttgggatca ttacaacggt cgcagctttt catcaggaat gcagcctaca
     1021 atcgtgtacc cagcaccagc cgtacgttgt ggacgatcct tgccccatcc acttctattc
     1081 taagtggtac atacgggtag gcgcgaggaa gagtgcacca ctcattgagt tgtgtgtgga
     1141 cgaggccggt tcaaaatccc caatccagta tatagatatt ggaaattaca ctgtctcctg
     1201 tctgcccttc accatcaact gccaagaacc taagctggga agcctcgttg tgagatgctc
     1261 tttttatgag gattttcttg aataccacga cgtccgtgtg gttctggact tcattgaaaa
     1321 cctttacttt cagtcgtacc cctatgacgt accagattat gcgtaaggat ccgcaggcct
     1381 ctgctagctt gactgactga gatacagcgt accttcagct cacagacatg ataagataca
     1441 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     1501 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     1561 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     1621 agtaaaacct ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct
     1681 tggggcctct aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta
     1741 ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
     1801 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
     1861 taccagaaat ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc
     1921 aagacattcc tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa
     1981 ctttcttccc tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc
     2041 aagtcagcta cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca
     2101 gtcaagttcg atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa
     2161 tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2221 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2281 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2341 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2401 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2461 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2521 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2581 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2641 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2701 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2761 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2821 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2881 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2941 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3001 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga
     3061 actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct
     3121 acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg
     3181 cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga
     3241 gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg
     3301 ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg
     3361 cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct gatccttccg cccggacgct
     3421 caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga
     3481 cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac
     3541 aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga
     3601 acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg
     3661 atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct
     3721 gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga
     3781 gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg
     3841 ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg
     3901 ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt
     3961 gagcccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg caccatctgg
     4021 ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg
     4081 cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg
     4141 tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc acctgccggt
     4201 aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct
     4261 cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct
     4321 ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg
     4381 ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt
     4441 tgggtcaata tgtaattttc agtgttagac ttgtaaattg tccgctaaat tctggccgtt
     4501 tttggctttt ttgttagaca acatgggtaa aaagcctgaa ctcaccgcga cgtctgtcga
     4561 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
     4621 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
     4681 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat ccgccgcgct
     4741 cccgattccg gaagtgcttg acattgggga gttcagcgag agcctgacct attgcatctc
     4801 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
     4861 gcagccggtc gcggaggcaa tggatgccat cgctgccgcc gatcttagcc agacgagcgg
     4921 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
     4981 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc
     5041 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg
     5101 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac
     5161 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat
     5221 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag
     5281 gcatccggag cttgcaggat cgccccggct ccgggcgtat atgctccgca ttggtcttga
     5341 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg
     5401 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag
     5461 aagcgccgcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg
     5521 ccccagcact cgtccgaggg caaaggaata agctagtatg taagcctagt cttagataat
     5581 aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg
     5641 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     5701 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     5761 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     5821 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     5881 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5941 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     6001 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     6061 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     6121 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     6181 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     6241 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     6301 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     6361 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6421 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     6481 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     6541 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6601 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6661 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6721 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     6781 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     6841 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     6901 agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt t
//
