LOCUS       sequence-295243-        4650 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 4650)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4650)
  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-m4133884
FEATURES             Location/Qualifiers
     source          1..4650
                     /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"
     CDS             join(2643..2711,2712..3503)
                     /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(2861..2880)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3516..3652
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3804..3875
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3880..3926)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3944..3992
                     /note="synthetic polyadenylation signal"
     enhancer        4073..4376
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4378..4580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4531..4551
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaaa tttcttgttt tcttaggaat catcacaact gtagctgcat ttcaccaaga
       61 atgtagttta cagtcatgta ctcaacatca accatatgta gttgatgacc cgtgtcctat
      121 tcacttctat tctaaatggt atattagagt aggagctaga aaatcagcac ctttaattga
      181 attgtgcgtg gatgaggctg gttctaaatc acccattcag tacatcgata tcggtaatta
      241 tacagtttcc tgtttacctt ttacaattaa ttgccaggaa cctaaattgg gtagtcttgt
      301 agtgcgttgt tcgttctatg aagacttttt agagtatcat gacgttcgtg ttgttttaga
      361 tttcatcgaa 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 gcgcctggtg
     2401 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag
     2461 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct
     2521 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt
     2581 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc
     2641 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2701 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2761 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2821 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2881 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2941 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3001 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3061 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3121 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3181 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag
     3241 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc
     3301 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3361 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3421 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3481 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     3541 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     3601 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga
     3661 atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag
     3721 ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt
     3781 ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc
     3841 tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg
     3901 tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca
     3961 cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga
     4021 tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     4081 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     4141 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     4201 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     4261 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     4321 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga
     4381 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     4441 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     4501 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     4561 aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac
     4621 catccactcg acacacccgc cagcggccgc
//
