LOCUS       sequence-312893-        4617 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 4617)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4617)
  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-m4133337
FEATURES             Location/Qualifiers
     source          1..4617
                     /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..1011
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1064..1185
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1075..1094)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1129..1148
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1300..1346
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1376..1512
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1557..1628
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1854..2442)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1934..1953)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3188..3256,3257..4048)
                     /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(3406..3425)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4061..4197
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4349..4420
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4425..4471)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4489..4537
                     /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 cagtcgaaag aaaccgcggc agctaagttc gagaggcagc acatggacag ttaagctagc
     1021 ttgactgact gagatacagc gtaccttcag ctcacagaca tgataagata cattgatgag
     1081 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     1141 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     1201 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     1261 ctctacaaat gtggtattgg cccatctcta tcggtatcgt agcataaccc cttggggcct
     1321 ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg gattgctatc taccggcatt
     1381 ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat
     1441 tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa
     1501 atttatcctt aaggtcgtca gctatcctgc aggcgatctc tcgatttcga tcaagacatt
     1561 cctttaatgg tcttttctgg acaccactag gggtcagaag tagttcatca aactttcttc
     1621 cctccctaat ctcattggtt accttgggct atcgaaactt aattaaccag tcaagtcagc
     1681 tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt cagtcaagtt
     1741 cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta aatcatgtga
     1801 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1861 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1921 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1981 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2041 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2101 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2161 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2221 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2281 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2341 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2401 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2461 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2521 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2581 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2641 atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc gaactctcca
     2701 aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg
     2761 aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt ggcagcgcct
     2821 atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga gagaagttca
     2881 acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc
     2941 tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg ttgcttggca
     3001 gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat attgtactca
     3061 agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt ctgatcggtc
     3121 aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat cagcaggagg
     3181 ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     3241 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     3301 gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     3361 tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     3421 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     3481 tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     3541 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     3601 aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3661 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3721 cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg aactacttac
     3781 tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg caggaccact
     3841 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     3901 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     3961 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4021 aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc agaaaaaaat
     4081 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
     4141 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaagat
     4201 cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg agcttacgcg
     4261 aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat cgtcagctta
     4321 cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac aggtatcttc
     4381 ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa acccctcaag
     4441 acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca cacaaaaaac
     4501 caacacacat ccatcttcga tggatagcga ttttattatc taactgctga tcgagtgtag
     4561 ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccg
//
