LOCUS       sequence-295390-        6862 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 6862)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6862)
  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-m4133651
FEATURES             Location/Qualifiers
     source          1..6862
                     /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             737..754
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    831..952
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(842..861)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     896..915
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1067..1113
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1143..1279
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1324..1395
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1621..2209)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1701..1720)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2709..3918
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3755..3774
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3922..4521
                     /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(3922..3941)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4418..4438
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4548..4596
                     /note="synthetic polyadenylation signal"
     CDS             join(4862..4930,4931..5722)
                     /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(5080..5099)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5735..5871
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6016..6087
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6092..6138)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6156..6204
                     /note="synthetic polyadenylation signal"
     enhancer        6285..6588
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6590..6792
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6743..6763
                     /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 gagtgacaaa atcattcatc taaccgatga
      421 ctcctttgat acagacgtgt tgaaggccga tggtgcgata ctggtagatt tctgggctga
      481 atggtgtggc ccgtgcaaga tgatcgcacc cattctggac gagatcgccg acgagtatca
      541 aggaaaatta actgtcgcta agcttaatat agatcagaac ccagggacgg cacctaaata
      601 cggcatcagg ggaataccta ccctcctgct ctttaagaat ggggaagttg ccgcgacaaa
      661 agtgggagcc ctgtctaagg gccagctaaa agagttcttg gacgctaacc tggcaggttc
      721 aggctcgggg cacatgcacc atcaccatca ccatagcagc gggctcgtgc cacgcggcta
      781 agctagcttg actgactgag atacagcgta ccttcagctc acagacatga taagatacat
      841 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
      901 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
      961 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     1021 gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc ataacccctt
     1081 ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat tgctatctac
     1141 cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     1201 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     1261 accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg atttcgatca
     1321 agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag ttcatcaaac
     1381 tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat taaccagtca
     1441 agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga attgcgtcag
     1501 tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt tcatttaaat
     1561 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1621 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1681 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1741 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1801 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1861 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1921 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     1981 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2041 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2101 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2161 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2221 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2281 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2341 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2401 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta aatttccgaa
     2461 ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc ttgcaggcta
     2521 cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct attgcttggc
     2581 agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat cacccgagag
     2641 aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat cgaaatcggg
     2701 gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc
     2761 gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc ccggacgctc
     2821 aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac
     2881 attttaggac gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca
     2941 ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa
     3001 cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga
     3061 tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg
     3121 ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag
     3181 agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact gggggttggg
     3241 gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg
     3301 gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc caaggtcttg
     3361 agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc accatctggg
     3421 gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg ttgcgggggc
     3481 ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt
     3541 cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca cctgccggta
     3601 ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc
     3661 ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg tcagtttctt
     3721 tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg aactatgcgc
     3781 tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt
     3841 gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt ctggccgttt
     3901 ttggcttttt tgttagacaa catgaccgag tacaagccca cggtgcgcct cgccacccgc
     3961 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     4021 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     4081 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     4141 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     4201 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga gggcctcctg
     4261 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     4321 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc cgccgagcgc
     4381 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     4441 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     4501 acccgcaagc ccggtgccta ggctagtatg taagcctagt cttagataat aaaatcgcta
     4561 tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt
     4621 accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc
     4681 cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg
     4741 gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta
     4801 taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg
     4861 catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4921 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4981 gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5041 cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5101 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     5161 ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5221 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5281 tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5341 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5401 gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc
     5461 ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5521 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5581 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5641 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5701 ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga
     5761 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     5821 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcgttta
     5881 aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg
     5941 ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc
     6001 gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg
     6061 tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg
     6121 ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca
     6181 tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa
     6241 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     6301 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     6361 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     6421 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     6481 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     6541 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt
     6601 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     6661 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     6721 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     6781 ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact
     6841 cgacacaccc gccagcggcc gc
//
