LOCUS       sequence-295867-        4599 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 4599)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4599)
  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-m4133269
FEATURES             Location/Qualifiers
     source          1..4599
                     /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..415
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    468..589
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(479..498)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     533..552
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      704..750
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      780..916
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    961..1032
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1258..1846)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1338..1357)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2592..2660,2661..3452)
                     /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(2810..2829)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3465..3601
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3753..3824
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3829..3875)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3893..3941
                     /note="synthetic polyadenylation signal"
     enhancer        4022..4325
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4327..4529
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4480..4500
                     /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 ttcagtcgta cccctatgac gtaccagatt atgcgtaagc
      421 tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga
      481 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
      541 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
      601 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
      661 aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg
      721 gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg
      781 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
      841 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
      901 agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga
      961 cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt
     1021 cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt
     1081 cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca
     1141 agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat
     1201 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1261 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1321 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1381 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1441 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1501 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1561 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1621 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1681 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1741 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1801 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1861 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1921 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     1981 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2041 acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc
     2101 tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct
     2161 ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc
     2221 gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag
     2281 ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg
     2341 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     2401 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     2461 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     2521 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     2581 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     2641 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     2701 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     2761 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     2821 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     2881 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     2941 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3001 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3061 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3121 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     3181 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     3241 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3301 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3361 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3421 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     3481 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     3541 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     3601 agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta
     3661 cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag
     3721 cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat
     3781 cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct
     3841 caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa
     3901 aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt
     3961 gtagccagat ctagtaatca attacggggt cattagttca tagcccatat atggagttcc
     4021 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     4081 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     4141 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     4201 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     4261 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     4321 ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     4381 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     4441 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     4501 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt
     4561 cgatcctacc atccactcga cacacccgcc agcggccgc
//
