LOCUS       sequence-317520-        6497 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 6497)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6497)
  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-m4134363
FEATURES             Location/Qualifiers
     source          1..6497
                     /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..984
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1037..1158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1048..1067)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1102..1121
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1273..1319
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1349..1485
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1530..1601
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1827..2415)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1907..1926)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2915..4124
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3961..3980
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4128..4727
                     /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(4128..4147)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4624..4644
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4754..4802
                     /note="synthetic polyadenylation signal"
     CDS             join(5068..5136,5137..5928)
                     /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(5286..5305)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5941..6077
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6229..6300
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6305..6351)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6369..6417
                     /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 cagtcgcacc atcaccatca ccattaagct agcttgactg actgagatac agcgtacctt
     1021 cagctcacag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     1081 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     1141 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     1201 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct
     1261 ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt
     1321 gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac
     1381 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     1441 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc
     1501 tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac
     1561 taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg
     1621 gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt
     1681 ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt
     1741 tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac
     1801 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1861 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1921 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1981 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2041 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2101 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2161 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2221 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2281 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2341 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2401 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2461 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2521 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2581 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2641 tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc
     2701 tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg
     2761 gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa
     2821 tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc
     2881 gagatccgag gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt tttggcgcct
     2941 cccgcgggcg cccccctcgt cacggcgagc gctgccacgt cagacgaagg gcgcaggagc
     3001 gtcctgatcc ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct
     3061 tagaacccca gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac
     3121 tggttttctt tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg
     3181 cggagggatc tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg
     3241 cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg
     3301 atcgtcactt ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc
     3361 gctcggtggg acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc
     3421 aaggttgccc tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc
     3481 ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg
     3541 gtgggcggca agaacccaag gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg
     3601 gtgagatggg ctgggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa
     3661 ctcggtttgt cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc
     3721 gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata
     3781 atgcagggtg gggccacctg ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc
     3841 agggttcggg cctagggtag gctctcctga atcgacaggc gccggacctc tggtgagggg
     3901 agggataagt gaggcgtcag tttctttggt cggttttatg tacctatctt cttaagtagc
     3961 tgaagctccg gttttgaact atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt
     4021 aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagacttgta
     4081 aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacaacatg accgagtaca
     4141 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     4201 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     4261 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     4321 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     4381 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     4441 cgcagcaaca gatggagggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     4501 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     4561 tccccggagt ggaggccgcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     4621 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     4681 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctaggct agtatgtaag
     4741 cctagtctta gataataaaa tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg
     4801 tgtgtaacgc taggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg
     4861 atccatatcg ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca
     4921 atcgtcagat attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga
     4981 tattgatagt ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa
     5041 gagacaggat cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt
     5101 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5161 aaagatgctg aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc
     5221 ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa
     5281 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5341 cgcatacact attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt
     5401 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5461 gcggccaact tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac
     5521 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5581 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta
     5641 ttaactggcg aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg
     5701 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5761 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5821 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5881 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt
     5941 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     6001 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     6061 cagaaattta tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc
     6121 cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct
     6181 cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca
     6241 ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc
     6301 aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt
     6361 gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc
     6421 taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc
     6481 ccatatatgg agttccg
//
