LOCUS       sequence-357055-        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-m4133893
FEATURES             Location/Qualifiers
     source          1..6993
                     /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    526..647
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(537..556)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     591..610
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      762..808
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      838..974
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1019..1090
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1316..1904)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1396..1415)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2404..3613
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3450..3469
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3617..4645
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4672..4720
                     /note="synthetic polyadenylation signal"
     CDS             join(4986..5054,5055..5846)
                     /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(5204..5223)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5859..5995
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6147..6218
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6223..6269)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6287..6335
                     /note="synthetic polyadenylation signal"
     enhancer        6416..6719
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6721..6923
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6874..6894
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaaa tttcttgttt tcttaggaat catcacaact gtagctgcat ttcaccaaga
       61 atgtagttta cagtcatgta ctcaacatca accatatgta gttgatgacc cgtgtcctat
      121 tcacttctat tctaaatggt atattagagt aggagctaga aaatcagcac ctttaattga
      181 attgtgcgtg gatgaggctg gttctaaatc acccattcag tacatcgata tcggtaatta
      241 tacagtttcc tgtttacctt ttacaattaa ttgccaggaa cctaaattgg gtagtcttgt
      301 agtgcgttgt tcgttctatg aagacttttt agagtatcat gacgttcgtg ttgttttaga
      361 tttcatcgaa aacctttact ttcagtcgat ggattataag gaccacgatg gagactacaa
      421 agatcatgac attgattaca aggacgacga tgacaaataa ggatccgcag gcctctgcta
      481 gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg
      541 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
      601 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
      661 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
      721 acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc
      781 ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca
      841 ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag
      901 attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag
      961 aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca
     1021 ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct
     1081 tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca
     1141 gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag
     1201 ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt
     1261 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1321 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1381 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1441 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1501 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1561 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     1621 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     1681 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     1741 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     1801 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     1861 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1921 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     1981 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2041 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2101 ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc
     2161 caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct
     2221 cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc
     2281 ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt
     2341 caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg
     2401 cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg
     2461 ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct tccgcccgga cgctcaggac
     2521 agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt
     2581 aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga
     2641 ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg
     2701 atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg
     2761 gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg
     2821 ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc
     2881 gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag
     2941 cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt
     3001 gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgagccc
     3061 ttcgctaatg cgggaaagct cttattcggg tgagatgggc tgggcaccat ctggggaccc
     3121 tgacgtgaag tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag
     3181 ttatggcggt gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt
     3241 cgtgacgtca cccgttctgt tggcttataa tgcagggtgg ggccacctgc cggtaggtgt
     3301 gcggtaggct tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa
     3361 tcgacaggcg ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc
     3421 ggttttatgt acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg
     3481 gttggcgagt gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc
     3541 aatatgtaat tttcagtgtt agacttgtaa attgtccgct aaattctggc cgtttttggc
     3601 ttttttgtta gacaacatgg gtaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt
     3661 tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc
     3721 tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc
     3781 cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatccgccg cgctcccgat
     3841 tccggaagtg cttgacattg gggagttcag cgagagcctg acctattgca tctcccgccg
     3901 tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc
     3961 ggtcgcggag gcaatggatg ccatcgctgc cgccgatctt agccagacga gcgggttcgg
     4021 cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat
     4081 tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt
     4141 cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct
     4201 cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt
     4261 cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt
     4321 ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc
     4381 ggagcttgca ggatcgcccc ggctccgggc gtatatgctc cgcattggtc ttgaccaact
     4441 ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga
     4501 cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc
     4561 cgccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag
     4621 cactcgtccg agggcaaagg aataagctag tatgtaagcc tagtcttaga taataaaatc
     4681 gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg
     4741 gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt
     4801 cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag
     4861 cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa
     4921 cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct
     4981 ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5041 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5101 gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5161 gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5221 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5281 acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5341 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5401 cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5461 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5521 cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc
     5581 tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc
     5641 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5701 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5761 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5821 gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc
     5881 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     5941 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc
     6001 cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa
     6061 cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc
     6121 tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt
     6181 ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac
     6241 ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca
     6301 acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc
     6361 agatctagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     6421 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     6481 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     6541 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     6601 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     6661 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc
     6721 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     6781 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     6841 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     6901 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc
     6961 taccatccac tcgacacacc cgccagcggc cgc
//
