LOCUS       sequence-295435-        4617 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 4617)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4617)
  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-m4133193
FEATURES             Location/Qualifiers
     source          1..4617
                     /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    486..607
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(497..516)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     551..570
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      722..768
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      798..934
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    979..1050
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1276..1864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1356..1375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2610..2678,2679..3470)
                     /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(2828..2847)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3483..3619
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3771..3842
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3847..3893)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3911..3959
                     /note="synthetic polyadenylation signal"
     enhancer        4040..4343
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4345..4547
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4498..4518
                     /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 agttaagcta gcttgactga ctgagataca gcgtaccttc agctcacaga
      481 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
      541 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
      601 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
      661 ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc
      721 gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc
      781 cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
      841 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
      901 atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc
      961 tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga
     1021 agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac
     1081 ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc
     1141 tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac
     1201 gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1261 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1321 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1381 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1441 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1501 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1561 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1621 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1681 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1741 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1801 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1861 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1921 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1981 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2041 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2101 atttaaattt ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat
     2161 ccatcttgca ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct
     2221 tggctattgc ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc
     2281 gggatcaccc gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg
     2341 aatatcgaaa tcggggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga
     2401 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc
     2461 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta
     2521 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc
     2581 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta
     2641 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2701 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca
     2761 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta
     2821 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     2881 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc
     2941 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3001 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc
     3061 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3121 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac
     3181 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg
     3241 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     3301 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     3361 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     3421 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag
     3481 gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     3541 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     3601 accagaaatt tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc
     3661 tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat
     3721 ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac
     3781 cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc
     3841 tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg
     3901 ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta
     3961 tctaactgct gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata
     4021 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     4081 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     4141 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     4201 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     4261 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     4321 tattagtcat cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat
     4381 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     4441 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     4501 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
     4561 gtcagatcag atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgc
//
