LOCUS       sequence-325524-        6506 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 6506)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6506)
  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-m4133405
FEATURES             Location/Qualifiers
     source          1..6506
                     /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"
     promoter        2346..3555
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3392..3411
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3559..4158
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3559..3578)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4055..4075
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4185..4233
                     /note="synthetic polyadenylation signal"
     CDS             join(4499..4567,4568..5359)
                     /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(4717..4736)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5372..5508
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5660..5731
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5736..5782)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5800..5848
                     /note="synthetic polyadenylation signal"
     enhancer        5929..6232
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6234..6436
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6387..6407
                     /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 cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag
     2401 cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg
     2461 acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt
     2521 ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc
     2581 gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc
     2641 cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt
     2701 gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag cgggctgctg
     2761 ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg tgtggagaga
     2821 ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg ggttgggggg
     2881 agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt gaggcgggct
     2941 gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa ggtcttgagc
     3001 ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctgggcacc atctggggac
     3061 cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg cgggggcggc
     3121 agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt
     3181 gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacct gccggtaggt
     3241 gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta ggctctcctg
     3301 aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca gtttctttgg
     3361 tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg
     3421 gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta atcatttggg
     3481 tcaatatgta attttcagtg ttagacttgt aaattgtccg ctaaattctg gccgtttttg
     3541 gcttttttgt tagacaacat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     3601 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     3661 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     3721 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     3781 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     3841 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaggg cctcctggcg
     3901 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     3961 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggccgc cgagcgcgcc
     4021 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     4081 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     4141 cgcaagcccg gtgcctaggc tagtatgtaa gcctagtctt agataataaa atcgctatcc
     4201 atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc
     4261 gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag
     4321 tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc
     4381 acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa
     4441 tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat
     4501 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4561 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg
     4621 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4681 agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4741 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4801 tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4861 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg
     4921 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4981 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5041 tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc
     5101 ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5161 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5221 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5281 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5341 actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc
     5401 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     5461 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga tcccgaatcg
     5521 tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt
     5581 ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg
     5641 cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta
     5701 gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta
     5761 gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca
     5821 tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta
     5881 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     5941 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     6001 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     6061 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     6121 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     6181 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tgctgatgcg
     6241 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     6301 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     6361 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     6421 ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga tcctaccatc
     6481 cactcgacac acccgccagc ggccgc
//
