LOCUS       sequence-295762-        7307 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 7307)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7307)
  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-m4134275
FEATURES             Location/Qualifiers
     source          1..7307
                     /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             737..754
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    847..968
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(858..877)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     912..931
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1083..1129
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1159..1295
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1340..1411
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1637..2225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1717..1736)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2725..3934
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3771..3790
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3938..4966
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4993..5041
                     /note="synthetic polyadenylation signal"
     CDS             join(5307..5375,5376..6167)
                     /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(5525..5544)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6180..6316
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6461..6532
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6537..6583)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6601..6649
                     /note="synthetic polyadenylation signal"
     enhancer        6730..7033
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7035..7237
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7188..7208
                     /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 ttcagtcgat gagtgacaaa atcattcatc taaccgatga
      421 ctcctttgat acagacgtgt tgaaggccga tggtgcgata ctggtagatt tctgggctga
      481 atggtgtggc ccgtgcaaga tgatcgcacc cattctggac gagatcgccg acgagtatca
      541 aggaaaatta actgtcgcta agcttaatat agatcagaac ccagggacgg cacctaaata
      601 cggcatcagg ggaataccta ccctcctgct ctttaagaat ggggaagttg ccgcgacaaa
      661 agtgggagcc ctgtctaagg gccagctaaa agagttcttg gacgctaacc tggcaggttc
      721 aggctcgggg cacatgcacc atcaccatca ccatagcagc gggctcgtgc cacgcggcta
      781 aggatccgca ggcctctgct agcttgactg actgagatac agcgtacctt cagctcacag
      841 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
      901 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
      961 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     1021 aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat
     1081 cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg
     1141 ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc
     1201 ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc
     1261 catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat
     1321 ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag
     1381 aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa
     1441 cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg
     1501 ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta
     1561 cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1621 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1681 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1741 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1801 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1861 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1921 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1981 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2041 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2101 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2161 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2221 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2281 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2341 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     2401 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     2461 catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga
     2521 tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc
     2581 ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat
     2641 cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag
     2701 gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg
     2761 cccccctcgt cacggcgagc gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc
     2821 ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct tagaacccca
     2881 gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac tggttttctt
     2941 tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg cggagggatc
     3001 tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt
     3061 tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt
     3121 ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc gctcggtggg
     3181 acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc
     3241 tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa
     3301 gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca
     3361 agaacccaag gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg gtgagatggg
     3421 ctgggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt
     3481 cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg
     3541 gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg
     3601 gggccacctg ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg
     3661 cctagggtag gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt
     3721 gaggcgtcag tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg
     3781 gttttgaact atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt
     3841 tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagacttgta aattgtccgc
     3901 taaattctgg ccgtttttgg cttttttgtt agacaacatg ggtaaaaagc ctgaactcac
     3961 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     4021 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
     4081 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
     4141 tgcatccgcc gcgctcccga ttccggaagt gcttgacatt ggggagttca gcgagagcct
     4201 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
     4261 actgcccgct gttctgcagc cggtcgcgga ggcaatggat gccatcgctg ccgccgatct
     4321 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     4381 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     4441 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     4501 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     4561 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     4621 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     4681 ctacttcgag cggaggcatc cggagcttgc aggatcgccc cggctccggg cgtatatgct
     4741 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     4801 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     4861 acaaatcgcc cgcagaagcg ccgccgtctg gaccgatggc tgtgtagaag tactcgccga
     4921 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaataagcta gtatgtaagc
     4981 ctagtcttag ataataaaat cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt
     5041 gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga
     5101 tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa
     5161 tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat
     5221 attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag
     5281 agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc
     5341 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5401 aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg
     5461 gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag
     5521 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     5581 gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta
     5641 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5701 cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca
     5761 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5821 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat
     5881 taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg
     5941 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6001 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6061 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6121 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg
     6181 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     6241 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     6301 agaaatttat ccttaagatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt
     6361 cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct
     6421 atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc
     6481 acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa
     6541 aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca
     6601 cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta tctaactgct
     6661 gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat
     6721 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     6781 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     6841 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     6901 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     6961 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     7021 cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     7081 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     7141 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     7201 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatcag
     7261 atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgc
//
