LOCUS       sequence-295217-        7934 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF3a protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7934)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7934)
  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-m4133679
FEATURES             Location/Qualifiers
     source          1..7934
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             830..850
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             884..961
                     /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             989..1009
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1474..1595
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1485..1504)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1539..1558
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1710..1756
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1786..1922
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1967..2038
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2264..2852)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2344..2363)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3352..4561
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4398..4417
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4565..5593
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5620..5668
                     /note="synthetic polyadenylation signal"
     CDS             join(5934..6002,6003..6794)
                     /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(6152..6171)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6807..6943
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7088..7159
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7164..7210)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7228..7276
                     /note="synthetic polyadenylation signal"
     enhancer        7357..7660
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7662..7864
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7815..7835
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7933..7934,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggat ttgtttatga gaatcttcac aattggaact gtaactttga agcaaggtga
       61 aatcaaggat gctactcctt cagattttgt tcgcgctact gcaacgatac cgatacaagc
      121 ctcactccct ttcggatggc ttattgttgg cgttgcactt cttgctgttt ttcagagcgc
      181 ttccaaaatc ataaccctca aaaagagatg gcaactagca ctctccaagg gtgttcactt
      241 tgtttgcaac ttgctgttgt tgtttgtaac agtttactca caccttttgc tcgttgctgc
      301 tggccttgaa gccccttttc tctatcttta tgctttagtc tacttcttgc agagtataaa
      361 ctttgtaaga ataataatga ggctttggct ttgctggaaa tgccgttcca aaaacccatt
      421 actttatgat gccaactatt ttctttgctg gcatactaat tgttacgact attgtatacc
      481 ttacaatagt gtaacttctt caattgtcat tacttcaggt gatggcacaa caagtcctat
      541 ttctgaacat gactaccaga ttggtggtta tactgaaaaa tgggaatctg gagtaaaaga
      601 ctgtgttgta ttacacagtt acttcacttc agactattac cagctgtact caactcaatt
      661 gagtacagac actggtgttg aacatgttac cttcttcatc tacaataaaa ttgttgatga
      721 gcctgaagaa catgtccaaa ttcacacaat cgacggttca tccggagttg ttaatccagt
      781 aatggaacca atttatgatg aaccgacgac gactactagc gtgcctttgg aaaaccttta
      841 ctttcagtcg ggacggcgaa tcccggggct gataaatccc tggaaaagga gatggaagaa
      901 gaactttatt gcagtgtctg cggctaaccg gttcaaaaag atcagttcaa gcggcgccct
      961 agattacgac attcctacga ccgcctccga gaatctctat ttccagggtg aatttggctt
     1021 ggcacaacat gacgaggccg tcgataacaa gtttaataaa gaacagcaga acgctttcta
     1081 tgagatcctt cacttaccaa atctgaacga agagcaacgc aacgctttca ttcagagcct
     1141 gaaagacgat ccctcgcagt ctgcaaatct cctggccgaa gccaagaagc tgaatgacgc
     1201 acaggctcca aaagttgata acaagtttaa caaggagcag cagaatgctt tctacgagat
     1261 attgcacctt cctaatctga atgaggaaca aagaaatgca tttatccagt ccctaaaaga
     1321 cgatccatca cagagtgcca acctgcttgc ggaagccaaa aagctcaatg atgcccaggc
     1381 cccgaaagta gacgcgaacc atcaataagg atccgcaggc ctctgctagc ttgactgact
     1441 gagatacagc gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa
     1501 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     1561 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     1621 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     1681 gtggtattgg cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt
     1741 cttgaggggt tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa
     1801 aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg
     1861 atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt
     1921 aaggtcgtca gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg
     1981 tcttttctgg acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat
     2041 ctcattggtt accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga
     2101 gatcgacttg tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc
     2161 cttgctattg cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc
     2221 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2281 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2341 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2401 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2461 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2521 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2581 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2641 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2701 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2761 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2821 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     2881 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2941 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3001 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3061 tctgtctatt tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt
     3121 cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat
     3181 cgcaagtctc ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt
     3241 attactccaa tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc
     3301 tcaatcccga tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc
     3361 gccgggtttt ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag
     3421 acgaagggcg caggagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg
     3481 ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt
     3541 gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc
     3601 ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa
     3661 ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt
     3721 gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct
     3781 ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt
     3841 agtctgggtc cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg
     3901 gcggctgttc ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa
     3961 acaaggtggg gggcatggtg ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg
     4021 ggaaagctct tattcgggtg agatgggctg ggcaccatct ggggaccctg acgtgaagtt
     4081 tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc
     4141 cgttgggcag tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc
     4201 cgttctgttg gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt
     4261 tctccgtcgc aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc
     4321 ggacctctgg tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac
     4381 ctatcttctt aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt
     4441 gttttgtgaa gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt
     4501 tcagtgttag acttgtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga
     4561 caacatgggt aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     4621 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     4681 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     4741 caaagatcgt tatgtttatc ggcactttgc atccgccgcg ctcccgattc cggaagtgct
     4801 tgacattggg gagttcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     4861 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     4921 aatggatgcc atcgctgccg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     4981 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     5041 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     5101 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     5161 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     5221 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     5281 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     5341 atcgccccgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     5401 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     5461 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgccg ccgtctggac
     5521 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     5581 ggcaaaggaa taagctagta tgtaagccta gtcttagata ataaaatcgc tatccatcga
     5641 agatggatgt gtgttggttt tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa
     5701 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     5761 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     5821 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     5881 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     5941 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     6001 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     6061 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     6121 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     6181 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     6241 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     6301 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     6361 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     6421 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     6481 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     6541 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6601 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6661 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6721 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6781 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     6841 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     6901 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcgtt taaactcgac
     6961 tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg
     7021 ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg
     7081 ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca
     7141 gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     7201 ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat
     7261 ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac
     7321 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     7381 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     7441 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     7501 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     7561 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     7621 ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt tttggcagta
     7681 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     7741 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     7801 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     7861 agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca ctcgacacac
     7921 ccgccagcgg ccgc
//
