LOCUS       sequence-295858-        6995 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 6995)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6995)
  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-m4133595
FEATURES             Location/Qualifiers
     source          1..6995
                     /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    535..656
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(546..565)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     600..619
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      771..817
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      847..983
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1028..1099
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1325..1913)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1405..1424)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2413..3622
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3459..3478
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3626..4654
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4681..4729
                     /note="synthetic polyadenylation signal"
     CDS             join(4995..5063,5064..5855)
                     /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(5213..5232)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5868..6004
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6149..6220
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6225..6271)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6289..6337
                     /note="synthetic polyadenylation signal"
     enhancer        6418..6721
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6723..6925
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6876..6896
                     /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 gatccgcagg
      481 cctctgctag cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat
      541 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
      601 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
      661 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
      721 gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc
      781 ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat
      841 ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
      901 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
      961 ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg
     1021 atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc
     1081 aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca
     1141 gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg
     1201 tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt
     1261 aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1321 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1381 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1441 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1501 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1561 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1621 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1681 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1741 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1801 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1861 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1921 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     1981 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2041 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     2101 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc
     2161 cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag
     2221 gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct
     2281 tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg
     2341 agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat
     2401 cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca
     2461 cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac
     2521 gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa
     2581 ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg
     2641 aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg
     2701 tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc
     2761 gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg
     2821 gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt
     2881 ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt
     2941 tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag
     3001 gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt
     3061 cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc
     3121 tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg
     3181 gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc
     3241 tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccacctgcc
     3301 ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc
     3361 tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt
     3421 tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat
     3481 gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc
     3541 atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc
     3601 gtttttggct tttttgttag acaacatggg taaaaagcct gaactcaccg cgacgtctgt
     3661 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg
     3721 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     3781 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catccgccgc
     3841 gctcccgatt ccggaagtgc ttgacattgg ggagttcagc gagagcctga cctattgcat
     3901 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt
     3961 tctgcagccg gtcgcggagg caatggatgc catcgctgcc gccgatctta gccagacgag
     4021 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat
     4081 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     4141 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     4201 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat
     4261 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa
     4321 catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg
     4381 gaggcatccg gagcttgcag gatcgccccg gctccgggcg tatatgctcc gcattggtct
     4441 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg
     4501 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg
     4561 cagaagcgcc gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg
     4621 acgccccagc actcgtccga gggcaaagga ataagctagt atgtaagcct agtcttagat
     4681 aataaaatcg ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag
     4741 gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg
     4801 cttggcagtc agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt
     4861 gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg
     4921 atcggtcaac gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag
     4981 caggaggctt tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5041 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5101 atcagttggg tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5161 agagttttcg ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5221 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5281 ctcagaatga cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga
     5341 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5401 ttctgacaac gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5461 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5521 gtgacaccac gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac
     5581 tacttactct agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag
     5641 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5701 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5761 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5821 ctgagatagg tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga
     5881 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     5941 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     6001 ttaagatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg agcttacgcg
     6061 aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat cgtcagctta
     6121 cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac aggtatcttc
     6181 ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa acccctcaag
     6241 acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca cacaaaaaac
     6301 caacacacat ccatcttcga tggatagcga ttttattatc taactgctga tcgagtgtag
     6361 ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     6421 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     6481 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     6541 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     6601 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     6661 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     6721 gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     6781 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     6841 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     6901 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat ctttgtcgat
     6961 cctaccatcc actcgacaca cccgccagcg gccgc
//
