LOCUS       sequence-295757-        6550 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 6550)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6550)
  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-m4133631
FEATURES             Location/Qualifiers
     source          1..6550
                     /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             389..466
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    519..640
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(530..549)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     584..603
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      755..801
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      831..967
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1012..1083
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1309..1897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1389..1408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2397..3606
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3443..3462
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3610..4209
                     /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(3610..3629)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4106..4126
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4236..4284
                     /note="synthetic polyadenylation signal"
     CDS             join(4550..4618,4619..5410)
                     /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(4768..4787)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5423..5559
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5704..5775
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5780..5826)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5844..5892
                     /note="synthetic polyadenylation signal"
     enhancer        5973..6276
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6278..6480
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6431..6451
                     /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 ttcagtcgaa gaggcgatgg aaaaagaatt ttatcgcggt
      421 gagtgcagcc aacagattca aaaagatatc ctcgtctggt gccctctaag ctagcttgac
      481 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
      541 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
      601 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
      661 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
      721 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
      781 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
      841 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
      901 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
      961 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
     1021 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
     1081 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
     1141 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
     1201 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
     1261 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1321 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1381 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1441 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1501 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1561 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1621 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1681 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1741 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1801 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1861 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1921 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1981 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2041 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2101 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     2161 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     2221 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     2281 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     2341 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc
     2401 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac
     2461 gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
     2521 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
     2581 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     2641 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     2701 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     2761 ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg
     2821 gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg
     2881 gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca
     2941 aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg
     3001 ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta
     3061 atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg
     3121 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
     3181 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
     3241 tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg tgtgcggtag
     3301 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
     3361 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
     3421 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
     3481 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
     3541 aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg
     3601 ttagacaaca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     3661 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     3721 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     3781 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     3841 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     3901 ttgagcggtt cccggctggc cgcgcagcaa cagatggagg gcctcctggc gccgcaccgg
     3961 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     4021 ggtctgggca gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc cggggtgccc
     4081 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     4141 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     4201 ggtgcctagg ctagtatgta agcctagtct tagataataa aatcgctatc catcgaagat
     4261 ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat
     4321 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     4381 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     4441 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     4501 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca
     4561 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4621 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta
     4681 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt
     4741 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4801 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc
     4861 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4921 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa
     4981 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5041 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5101 ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5161 gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5221 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5281 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5341 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5401 gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     5461 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     5521 cttccatacg tgtcctcctt accagaaatt tatccttaag atcgtttaaa ctcgactctg
     5581 gctctatcga atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg
     5641 tcgggcatcg aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc
     5701 cgacatcttg gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag
     5761 cttcagctac ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt
     5821 atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat
     5881 agcgatttta ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg
     5941 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     6001 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     6061 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     6121 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     6181 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     6241 cagtacatct acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc
     6301 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     6361 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     6421 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     6481 ggtttagtga accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc
     6541 cagcggccgc
//
