LOCUS       sequence-319581-        6548 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 6548)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6548)
  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-m4133996
FEATURES             Location/Qualifiers
     source          1..6548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        321..523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     474..494
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             961..981
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             985..1050
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1103..1224
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1114..1133)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1168..1187
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1339..1385
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1415..1551
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1596..1667
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1893..2481)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1973..1992)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2981..4190
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4027..4046
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4194..4793
                     /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(4194..4213)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4690..4710
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4820..4868
                     /note="synthetic polyadenylation signal"
     CDS             join(5134..5202,5203..5994)
                     /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(5352..5371)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6007..6143
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6295..6366
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6371..6417)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6435..6483
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
       61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      301 agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      541 gatcagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc cgccaccatg
      601 aaatttcttg ttttcttagg aatcatcaca actgtagctg catttcacca agaatgtagt
      661 ttacagtcat gtactcaaca tcaaccatat gtagttgatg acccgtgtcc tattcacttc
      721 tattctaaat ggtatattag agtaggagct agaaaatcag cacctttaat tgaattgtgc
      781 gtggatgagg ctggttctaa atcacccatt cagtacatcg atatcggtaa ttatacagtt
      841 tcctgtttac cttttacaat taattgccag gaacctaaat tgggtagtct tgtagtgcgt
      901 tgttcgttct atgaagactt tttagagtat catgacgttc gtgttgtttt agatttcatc
      961 gaaaaccttt actttcagtc gatggattat aaggaccacg atggagacta caaagatcat
     1021 gacattgatt acaaggacga cgatgacaaa taagctagct tgactgactg agatacagcg
     1081 taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac cacaactaga
     1141 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     1201 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     1261 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtattggc
     1321 ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     1381 ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa aaatgcctga
     1441 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     1501 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag
     1561 ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt cttttctgga
     1621 caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc tcattggtta
     1681 ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag atcgacttgt
     1741 ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc ttgctattgc
     1801 acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc
     1861 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1921 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1981 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2041 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2101 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2161 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2221 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2281 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2341 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2401 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2461 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2521 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2581 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2641 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2701 cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc ggaaaatctt
     2761 caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc gcaagtctct
     2821 tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta ttactccaat
     2881 cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct caatcccgat
     2941 ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggcctccgcg ccgggttttg
     3001 gcgcctcccg cgggcgcccc cctcgtcacg gcgagcgctg ccacgtcaga cgaagggcgc
     3061 aggagcgtcc tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact
     3121 cggccttaga accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta
     3181 gggcactggt tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg
     3241 attctgcgga gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg
     3301 gtgtggcaca gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc
     3361 gctgtgatcg tcacttggtg agtagcgggc tgctgggctg gccggggctt tcgtggccgc
     3421 cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc
     3481 gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg cggctgttcc
     3541 cgagtcttga atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg
     3601 ggcatggtgg gcggcaagaa cccaaggtct tgagcccttc gctaatgcgg gaaagctctt
     3661 attcgggtga gatgggctgg gcaccatctg gggaccctga cgtgaagttt gtcactgact
     3721 ggagaactcg gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt
     3781 gcacccgtac ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg
     3841 cttataatgc agggtggggc cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca
     3901 ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt
     3961 gaggggaggg ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta
     4021 agtagctgaa gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag
     4081 ttttttaggc accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga
     4141 cttgtaaatt gtccgctaaa ttctggccgt ttttggcttt tttgttagac aacatgaccg
     4201 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     4261 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     4321 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     4381 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     4441 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     4501 tggccgcgca gcaacagatg gagggcctcc tggcgccgca ccggcccaag gagcccgcgt
     4561 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     4621 tcgtgctccc cggagtggag gccgccgagc gcgccggggt gcccgccttc ctggagacct
     4681 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     4741 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc taggctagta
     4801 tgtaagccta gtcttagata ataaaatcgc tatccatcga agatggatgt gtgttggttt
     4861 tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc
     4921 tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga
     4981 agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa
     5041 cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc
     5101 tatcaagaga caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc
     5161 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5221 aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc
     5281 aacagcggta agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact
     5341 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5401 ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag
     5461 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5521 aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt
     5581 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5641 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt
     5701 aaactattaa ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg
     5761 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5821 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5881 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5941 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt
     6001 ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
     6061 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
     6121 ccttaccaga aatttatcct taagatcccg aatcgtttaa actcgactct ggctctatcg
     6181 aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc
     6241 gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt
     6301 ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta
     6361 cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca
     6421 atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt
     6481 attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt
     6541 catagccc
//
