LOCUS       sequence-295745-        7472 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 7472)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7472)
  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-m4133667
FEATURES             Location/Qualifiers
     source          1..7472
                     /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             422..499
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             527..547
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1012..1133
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1023..1042)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1077..1096
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1248..1294
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1324..1460
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1505..1576
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1802..2390)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1882..1901)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2890..4099
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3936..3955
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4103..5131
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5158..5206
                     /note="synthetic polyadenylation signal"
     CDS             join(5472..5540,5541..6332)
                     /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(5690..5709)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6345..6481
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6626..6697
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6702..6748)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6766..6814
                     /note="synthetic polyadenylation signal"
     enhancer        6895..7198
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7200..7402
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7353..7373
                     /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 ttcagtcggg acggcgaatc ccggggctga taaatccctg
      421 gaaaaggaga tggaagaaga actttattgc agtgtctgcg gctaaccggt tcaaaaagat
      481 cagttcaagc ggcgccctag attacgacat tcctacgacc gcctccgaga atctctattt
      541 ccagggtgaa tttggcttgg cacaacatga cgaggccgtc gataacaagt ttaataaaga
      601 acagcagaac gctttctatg agatccttca cttaccaaat ctgaacgaag agcaacgcaa
      661 cgctttcatt cagagcctga aagacgatcc ctcgcagtct gcaaatctcc tggccgaagc
      721 caagaagctg aatgacgcac aggctccaaa agttgataac aagtttaaca aggagcagca
      781 gaatgctttc tacgagatat tgcaccttcc taatctgaat gaggaacaaa gaaatgcatt
      841 tatccagtcc ctaaaagacg atccatcaca gagtgccaac ctgcttgcgg aagccaaaaa
      901 gctcaatgat gcccaggccc cgaaagtaga cgcgaaccat caataaggat ccgcaggcct
      961 ctgctagctt gactgactga gatacagcgt accttcagct cacagacatg ataagataca
     1021 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     1081 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     1141 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     1201 agtaaaacct ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct
     1261 tggggcctct aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta
     1321 ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
     1381 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
     1441 taccagaaat ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc
     1501 aagacattcc tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa
     1561 ctttcttccc tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc
     1621 aagtcagcta cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca
     1681 gtcaagttcg atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa
     1741 tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1801 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1861 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1921 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1981 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2041 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2101 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2161 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2221 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2281 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2341 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2401 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2461 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2521 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2581 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga
     2641 actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct
     2701 acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg
     2761 cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga
     2821 gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg
     2881 ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg
     2941 cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct gatccttccg cccggacgct
     3001 caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga
     3061 cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac
     3121 aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga
     3181 acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg
     3241 atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct
     3301 gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga
     3361 gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg
     3421 ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg
     3481 ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt
     3541 gagcccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg caccatctgg
     3601 ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg
     3661 cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg
     3721 tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc acctgccggt
     3781 aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct
     3841 cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct
     3901 ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg
     3961 ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt
     4021 tgggtcaata tgtaattttc agtgttagac ttgtaaattg tccgctaaat tctggccgtt
     4081 tttggctttt ttgttagaca acatgggtaa aaagcctgaa ctcaccgcga cgtctgtcga
     4141 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
     4201 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
     4261 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat ccgccgcgct
     4321 cccgattccg gaagtgcttg acattgggga gttcagcgag agcctgacct attgcatctc
     4381 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
     4441 gcagccggtc gcggaggcaa tggatgccat cgctgccgcc gatcttagcc agacgagcgg
     4501 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
     4561 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc
     4621 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg
     4681 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac
     4741 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat
     4801 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag
     4861 gcatccggag cttgcaggat cgccccggct ccgggcgtat atgctccgca ttggtcttga
     4921 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg
     4981 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag
     5041 aagcgccgcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg
     5101 ccccagcact cgtccgaggg caaaggaata agctagtatg taagcctagt cttagataat
     5161 aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg
     5221 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     5281 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     5341 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     5401 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     5461 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5521 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5581 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5641 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5701 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5761 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5821 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5881 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5941 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6001 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     6061 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     6121 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6181 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6241 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6301 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     6361 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     6421 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     6481 agatcgttta aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac
     6541 agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct
     6601 ttttggcagc gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc
     6661 cctctagtgg tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc
     6721 cgtttagagg ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa
     6781 cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca
     6841 gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     6901 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     6961 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     7021 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     7081 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     7141 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct
     7201 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     7261 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     7321 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     7381 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct
     7441 accatccact cgacacaccc gccagcggcc gc
//
