LOCUS       sequence-295354-        6962 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 6962)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6962)
  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-m4133633
FEATURES             Location/Qualifiers
     source          1..6962
                     /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..433
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    502..623
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(513..532)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     567..586
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      738..784
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      814..950
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    995..1066
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1292..1880)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1372..1391)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2380..3589
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3426..3445
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3593..4621
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4648..4696
                     /note="synthetic polyadenylation signal"
     CDS             join(4962..5030,5031..5822)
                     /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(5180..5199)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5835..5971
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6116..6187
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6192..6238)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6256..6304
                     /note="synthetic polyadenylation signal"
     enhancer        6385..6688
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6690..6892
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6843..6863
                     /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 ttcagtcgaa agaaaccgcg gcagctaagt tcgagaggca
      421 gcacatggac agttaaggat ccgcaggcct ctgctagctt gactgactga gatacagcgt
      481 accttcagct cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
      541 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
      601 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
      661 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc
      721 catctctatc ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt
      781 tttgtgcccc tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat
      841 gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc
      901 caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc
      961 tatcctgcag gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac
     1021 accactaggg gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac
     1081 cttgggctat cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc
     1141 tgggtttcga ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca
     1201 cccgttctcc gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca
     1261 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1321 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1381 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1441 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1501 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1561 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1621 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     1681 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     1741 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1801 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1861 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     1921 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     1981 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2041 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2101 gttcatccat agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc
     2161 aaacctttcg tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt
     2221 ggccggcctt gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc
     2281 ccgaatatcc gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc
     2341 tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg
     2401 cgcctcccgc gggcgccccc ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca
     2461 ggagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc
     2521 ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag
     2581 ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga
     2641 ttctgcggag ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg
     2701 tgtggcacag ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg
     2761 ctgtgatcgt cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc
     2821 gggccgctcg gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg
     2881 cgagcaaggt tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc
     2941 gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg
     3001 gcatggtggg cggcaagaac ccaaggtctt gagcccttcg ctaatgcggg aaagctctta
     3061 ttcgggtgag atgggctggg caccatctgg ggaccctgac gtgaagtttg tcactgactg
     3121 gagaactcgg tttgtcgtct gttgcggggg cggcagttat ggcggtgccg ttgggcagtg
     3181 cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc
     3241 ttataatgca gggtggggcc acctgccggt aggtgtgcgg taggcttttc tccgtcgcag
     3301 gacgcagggt tcgggcctag ggtaggctct cctgaatcga caggcgccgg acctctggtg
     3361 aggggaggga taagtgaggc gtcagtttct ttggtcggtt ttatgtacct atcttcttaa
     3421 gtagctgaag ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt
     3481 tttttaggca ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc agtgttagac
     3541 ttgtaaattg tccgctaaat tctggccgtt tttggctttt ttgttagaca acatgggtaa
     3601 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     3661 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     3721 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     3781 tgtttatcgg cactttgcat ccgccgcgct cccgattccg gaagtgcttg acattgggga
     3841 gttcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     3901 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcaa tggatgccat
     3961 cgctgccgcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     4021 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     4081 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     4141 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     4201 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     4261 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     4321 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccccggct
     4381 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
     4441 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     4501 gactgtcggg cgtacacaaa tcgcccgcag aagcgccgcc gtctggaccg atggctgtgt
     4561 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
     4621 agctagtatg taagcctagt cttagataat aaaatcgcta tccatcgaag atggatgtgt
     4681 gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg atcctctcag
     4741 tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg
     4801 acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc
     4861 tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg
     4921 caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc
     4981 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5041 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac
     5101 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga
     5161 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5221 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca
     5281 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5341 tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa
     5401 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5461 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5521 ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag
     5581 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5641 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5701 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5761 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5821 aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     5881 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     5941 cgtgtcctcc ttaccagaaa tttatcctta agatcgttta aactcgactc tggctctatc
     6001 gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat
     6061 cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct cccgacatct
     6121 tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc agcttcagct
     6181 acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctatc
     6241 aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg atagcgattt
     6301 tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg ggtcattagt
     6361 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     6421 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     6481 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     6541 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     6601 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     6661 ctacgtatta gtcatcgcta ttaccatgct gatgcggttt tggcagtaca tcaatgggcg
     6721 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     6781 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     6841 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
     6901 gaaccgtcag atcagatctt tgtcgatcct accatccact cgacacaccc gccagcggcc
     6961 gc
//
