LOCUS       sequence-311991-        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-m4133323
FEATURES             Location/Qualifiers
     source          1..6506
                     /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             967..993
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1046..1167
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1057..1076)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1111..1130
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1282..1328
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1358..1494
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1539..1610
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1836..2424)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1916..1935)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2924..4133
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3970..3989
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4137..4736
                     /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(4137..4156)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4633..4653
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4763..4811
                     /note="synthetic polyadenylation signal"
     CDS             join(5077..5145,5146..5937)
                     /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(5295..5314)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5950..6086
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6238..6309
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6314..6360)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6378..6426
                     /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 tcttgttttc
      601 ttaggaatca tcacaactgt agctgcattt caccaagaat gtagtttaca gtcatgtact
      661 caacatcaac catatgtagt tgatgacccg tgtcctattc acttctattc taaatggtat
      721 attagagtag gagctagaaa atcagcacct ttaattgaat tgtgcgtgga tgaggctggt
      781 tctaaatcac ccattcagta catcgatatc ggtaattata cagtttcctg tttacctttt
      841 acaattaatt gccaggaacc taaattgggt agtcttgtag tgcgttgttc gttctatgaa
      901 gactttttag agtatcatga cgttcgtgtt gttttagatt tcatcgaaaa cctttacttt
      961 cagtcgtacc cctatgacgt accagattat gcgtaagcta gcttgactga ctgagataca
     1021 gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca aaccacaact
     1081 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     1141 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     1201 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatt
     1261 ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg gtcttgaggg
     1321 gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga aaaaaatgcc
     1381 tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct
     1441 tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt
     1501 cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat ggtcttttct
     1561 ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta atctcattgg
     1621 ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc gagatcgact
     1681 tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg tccttgctat
     1741 tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag
     1801 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1861 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1921 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     1981 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2041 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2101 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2161 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2221 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2281 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2341 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2401 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2461 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2521 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2581 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2641 tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc gtcggaaaat
     2701 cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact atcgcaagtc
     2761 tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag gtattactcc
     2821 aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat cctcaatccc
     2881 gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggcctcc gcgccgggtt
     2941 ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg ctgccacgtc agacgaaggg
     3001 cgcaggagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc tgctcataag
     3061 actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac ttgggtgact
     3121 ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag tcccttctcg
     3181 gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat aaggacgcgc
     3241 cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg
     3301 atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc
     3361 cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct gtagtctggg
     3421 tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa tggcggctgt
     3481 tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg
     3541 gggggcatgg tgggcggcaa gaacccaagg tcttgagccc ttcgctaatg cgggaaagct
     3601 cttattcggg tgagatgggc tgggcaccat ctggggaccc tgacgtgaag tttgtcactg
     3661 actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt gccgttgggc
     3721 agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt
     3781 tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct tttctccgtc
     3841 gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg ccggacctct
     3901 ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt acctatcttc
     3961 ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg
     4021 aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat tttcagtgtt
     4081 agacttgtaa attgtccgct aaattctggc cgtttttggc ttttttgtta gacaacatga
     4141 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
     4201 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
     4261 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     4321 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     4381 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     4441 ggctggccgc gcagcaacag atggagggcc tcctggcgcc gcaccggccc aaggagcccg
     4501 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     4561 ccgtcgtgct ccccggagtg gaggccgccg agcgcgccgg ggtgcccgcc ttcctggaga
     4621 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     4681 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctaggcta
     4741 gtatgtaagc ctagtcttag ataataaaat cgctatccat cgaagatgga tgtgtgttgg
     4801 ttttttgtgt gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga
     4861 gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca
     4921 ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt
     4981 aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac
     5041 atctatcaag agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc
     5101 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5161 gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat
     5221 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc
     5281 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5341 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa
     5401 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5461 gataacactg cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct
     5521 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5581 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg
     5641 cgtaaactat taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg
     5701 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5761 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5821 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5881 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg
     5941 attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc
     6001 cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt
     6061 cctccttacc agaaatttat ccttaagatc ccgaatcgtt taaactcgac tctggctcta
     6121 tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc
     6181 atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat
     6241 cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag
     6301 ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta
     6361 tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat
     6421 tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac ggggtcatta
     6481 gttcatagcc catatatgga gttccg
//
