LOCUS       sequence-320320-        4599 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 4599)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4599)
  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-m4133269
FEATURES             Location/Qualifiers
     source          1..4599
                     /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"
     CDS             join(3170..3238,3239..4030)
                     /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(3388..3407)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4043..4179
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4331..4402
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4407..4453)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4471..4519
                     /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 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 cctggtgtac cgagaacgat
     2941 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     3001 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     3061 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     3121 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca
     3181 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3241 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta
     3301 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt
     3361 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3421 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc
     3481 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3541 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa
     3601 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3661 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3721 ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3781 gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3841 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     3901 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3961 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4021 gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     4081 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     4141 cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc gtttaaactc
     4201 gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat
     4261 ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg
     4321 cggctcccga catcttggac cattagctcc acaggtatct tcttccctct agtggtcata
     4381 acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca
     4441 aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc
     4501 gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat
     4561 tacggggtca ttagttcata gcccatatat ggagttccg
//
