LOCUS       sequence-295572-        4641 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 4641)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4641)
  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-m4134171
FEATURES             Location/Qualifiers
     source          1..4641
                     /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             392..457
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    510..631
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(521..540)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     575..594
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      746..792
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      822..958
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1003..1074
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1300..1888)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1380..1399)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2634..2702,2703..3494)
                     /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(2852..2871)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3507..3643
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3795..3866
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3871..3917)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3935..3983
                     /note="synthetic polyadenylation signal"
     enhancer        4064..4367
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4369..4571
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4522..4542
                     /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 ttcagtcgat ggattataag gaccacgatg gagactacaa
      421 agatcatgac attgattaca aggacgacga tgacaaataa gctagcttga ctgactgaga
      481 tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac
      541 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      601 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
      661 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
      721 tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg
      781 aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa
      841 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
      901 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg
      961 tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt
     1021 ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca
     1081 ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc
     1141 gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg
     1201 ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca
     1261 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1321 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1381 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1441 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1501 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1561 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1621 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1681 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1741 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1801 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1861 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1921 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     1981 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2041 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2101 tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga
     2161 aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca
     2221 agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta
     2281 ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa
     2341 tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa
     2401 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     2461 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     2521 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     2581 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     2641 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2701 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     2761 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2821 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     2881 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2941 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3001 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     3061 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3121 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3181 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     3241 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3301 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3361 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3421 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3481 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     3541 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     3601 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa
     3661 actcgactct ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc
     3721 tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg
     3781 atcgcggctc ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt
     3841 cataacagca gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc
     3901 cccaaggggt tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat
     3961 cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat
     4021 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     4081 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     4141 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     4201 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     4261 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     4321 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt
     4381 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     4441 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     4501 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     4561 taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc
     4621 gacacacccg ccagcggccg c
//
