LOCUS       sequence-295464-        5127 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 5127)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5127)
  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-m4133911
FEATURES             Location/Qualifiers
     source          1..5127
                     /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             422..499
                     /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"
     CDS             527..547
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    996..1117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1007..1026)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1061..1080
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1232..1278
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1308..1444
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1489..1560
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1786..2374)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1866..1885)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3120..3188,3189..3980)
                     /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(3338..3357)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3993..4129
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4281..4352
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4357..4403)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4421..4469
                     /note="synthetic polyadenylation signal"
     enhancer        4550..4853
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4855..5057
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5008..5028
                     /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 ttcagtcggg acggcgaatc ccggggctga taaatccctg
      421 gaaaaggaga tggaagaaga actttattgc agtgtctgcg gctaaccggt tcaaaaagat
      481 cagttcaagc ggcgccctag attacgacat tcctacgacc gcctccgaga atctctattt
      541 ccagggtgaa tttggcttgg cacaacatga cgaggccgtc gataacaagt ttaataaaga
      601 acagcagaac gctttctatg agatccttca cttaccaaat ctgaacgaag agcaacgcaa
      661 cgctttcatt cagagcctga aagacgatcc ctcgcagtct gcaaatctcc tggccgaagc
      721 caagaagctg aatgacgcac aggctccaaa agttgataac aagtttaaca aggagcagca
      781 gaatgctttc tacgagatat tgcaccttcc taatctgaat gaggaacaaa gaaatgcatt
      841 tatccagtcc ctaaaagacg atccatcaca gagtgccaac ctgcttgcgg aagccaaaaa
      901 gctcaatgat gcccaggccc cgaaagtaga cgcgaaccat caataagcta gcttgactga
      961 ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca
     1021 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     1081 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     1141 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     1201 atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg
     1261 gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga
     1321 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     1381 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     1441 ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat
     1501 ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta
     1561 atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc
     1621 gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg
     1681 tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg
     1741 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1801 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1861 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1921 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1981 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2041 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2101 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2161 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2221 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2281 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2341 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2401 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2461 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2521 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2581 gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc
     2641 gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact
     2701 atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag
     2761 gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat
     2821 cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggtgtac
     2881 cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca
     2941 gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt
     3001 cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata
     3061 atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca
     3121 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3181 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc
     3241 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3301 aagaacgctt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3361 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3421 ttgagtattc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3481 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg
     3541 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3601 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3661 ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt
     3721 cccggcaaca gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3781 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3841 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3901 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3961 cactgattaa gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg
     4021 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     4081 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc
     4141 gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg
     4201 tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg
     4261 gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct tcttccctct
     4321 agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt
     4381 agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac
     4441 atccatcttc gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct
     4501 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     4561 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     4621 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     4681 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     4741 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     4801 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc
     4861 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     4921 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     4981 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     5041 tctatataag cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat
     5101 ccactcgaca cacccgccag cggccgc
//
