LOCUS       sequence-317900-        6993 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 6993)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6993)
  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-m4133655
FEATURES             Location/Qualifiers
     source          1..6993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             946..966
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             970..1035
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1104..1225
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1115..1134)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1169..1188
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1340..1386
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1416..1552
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1597..1668
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1894..2482)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1974..1993)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2982..4191
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4028..4047
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4195..5223
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5250..5298
                     /note="synthetic polyadenylation signal"
     CDS             join(5564..5632,5633..6424)
                     /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(5782..5801)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6437..6573
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6725..6796
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6801..6847)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6865..6913
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgaaatt cttagtgttt
      601 cttgggatca ttacaacggt cgcagctttt catcaggaat gcagcctaca atcgtgtacc
      661 cagcaccagc cgtacgttgt ggacgatcct tgccccatcc acttctattc taagtggtac
      721 atacgggtag gcgcgaggaa gagtgcacca ctcattgagt tgtgtgtgga cgaggccggt
      781 tcaaaatccc caatccagta tatagatatt ggaaattaca ctgtctcctg tctgcccttc
      841 accatcaact gccaagaacc taagctggga agcctcgttg tgagatgctc tttttatgag
      901 gattttcttg aataccacga cgtccgtgtg gttctggact tcattgaaaa cctttacttt
      961 cagtcgatgg attataagga ccacgatgga gactacaaag atcatgacat tgattacaag
     1021 gacgacgatg acaaataagg atccgcaggc ctctgctagc ttgactgact gagatacagc
     1081 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1141 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1201 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1261 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg
     1321 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1381 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg
     1441 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     1501 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca
     1561 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg
     1621 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt
     1681 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg
     1741 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg
     1801 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc
     1861 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     1921 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     1981 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2041 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2101 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2161 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2221 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2281 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2341 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2401 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2461 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2521 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2581 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2641 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2701 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct
     2761 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc
     2821 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa
     2881 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga
     2941 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt
     3001 ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg
     3061 caggagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac
     3121 tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct
     3181 agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc
     3241 gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg
     3301 ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat
     3361 cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg
     3421 ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc
     3481 cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc
     3541 ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg
     3601 gggcatggtg ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg ggaaagctct
     3661 tattcgggtg agatgggctg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac
     3721 tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag
     3781 tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg
     3841 gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc
     3901 aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc ggacctctgg
     3961 tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt
     4021 aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa
     4081 gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt tcagtgttag
     4141 acttgtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga caacatgggt
     4201 aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc
     4261 gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta
     4321 ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt
     4381 tatgtttatc ggcactttgc atccgccgcg ctcccgattc cggaagtgct tgacattggg
     4441 gagttcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa
     4501 gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc aatggatgcc
     4561 atcgctgccg ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc
     4621 ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac
     4681 tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg
     4741 atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc
     4801 aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg
     4861 ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt
     4921 atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccccgg
     4981 ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc
     5041 aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc
     5101 gggactgtcg ggcgtacaca aatcgcccgc agaagcgccg ccgtctggac cgatggctgt
     5161 gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa
     5221 taagctagta tgtaagccta gtcttagata ataaaatcgc tatccatcga agatggatgt
     5281 gtgttggttt tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa cgatcctctc
     5341 agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg
     5401 ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga
     5461 tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt
     5521 tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta ttcaacattt
     5581 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5641 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg gttacatcga
     5701 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gctttccaat
     5761 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5821 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt
     5881 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5941 catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct
     6001 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     6061 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     6121 aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc aacagttgat
     6181 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6241 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6301 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6361 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6421 gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
     6481 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
     6541 tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa actcgactct
     6601 ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc
     6661 gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc
     6721 ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca
     6781 gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
     6841 tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga
     6901 tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg
     6961 gtcattagtt catagcccat atatggagtt ccg
//
