LOCUS       sequence-295245-        4614 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 4614)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4614)
  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-m4134020
FEATURES             Location/Qualifiers
     source          1..4614
                     /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    483..604
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(494..513)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     548..567
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      719..765
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      795..931
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    976..1047
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1273..1861)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1353..1372)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2607..2675,2676..3467)
                     /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(2825..2844)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3480..3616
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3768..3839
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3844..3890)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3908..3956
                     /note="synthetic polyadenylation signal"
     enhancer        4037..4340
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4342..4544
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4495..4515
                     /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 taagctagct tgactgactg agatacagcg taccttcagc tcacagacat
      481 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
      541 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
      601 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
      661 tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta
      721 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg
      781 attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
      841 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
      901 cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct
      961 cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt
     1021 agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta
     1081 attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca
     1141 gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag
     1201 tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1261 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1321 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1381 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1441 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1501 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1561 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1621 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1681 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1741 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1801 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     1861 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     1921 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     1981 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2041 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt
     2101 taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca
     2161 tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct tgcgccttgg
     2221 ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc cgagatcggg
     2281 atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga tccgaggaat
     2341 atcgaaatcg gggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga
     2401 tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa
     2461 tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat
     2521 attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag
     2581 agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc
     2641 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2701 aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg
     2761 gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag
     2821 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     2881 gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta
     2941 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3001 cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca
     3061 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3121 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat
     3181 taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg
     3241 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3301 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3361 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3421 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg
     3481 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     3541 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     3601 agaaatttat ccttaagatc ccgaatcgtt taaactcgac tctggctcta tcgaatctcc
     3661 gtcgtttcga gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc
     3721 gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat
     3781 tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca
     3841 attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg
     3901 cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct
     3961 aactgctgat cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc
     4021 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     4081 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     4141 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     4201 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     4261 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     4321 tagtcatcgc tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc
     4381 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     4441 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     4501 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     4561 agatcagatc tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgc
//
