LOCUS       sequence-307624-        6983 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 6983)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6983)
  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-m4134028
FEATURES             Location/Qualifiers
     source          1..6983
                     /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             851..892
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    945..1066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(956..975)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1010..1029
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1181..1227
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1257..1393
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1438..1509
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1735..2323)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1815..1834)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2823..4032
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3869..3888
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4036..4635
                     /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(4036..4055)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4532..4552
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4662..4710
                     /note="synthetic polyadenylation signal"
     CDS             join(4976..5044,5045..5836)
                     /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(5194..5213)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5849..5985
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6137..6208
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6213..6259)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6277..6325
                     /note="synthetic polyadenylation signal"
     enhancer        6406..6709
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6711..6913
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6864..6884
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6982..6983,1..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 gggaaaccta tacccaatcc acttctcggc ttagattcta cctaagctag
      901 cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat acattgatga
      961 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     1021 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1081 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     1141 cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc
     1201 tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat
     1261 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     1321 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     1381 aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat
     1441 tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt
     1501 ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag
     1561 ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt
     1621 tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg
     1681 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1741 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1801 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1861 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1921 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1981 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2041 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2101 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2161 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2221 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2281 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2341 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2401 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2461 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2521 tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc
     2581 aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc
     2641 gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc
     2701 tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc
     2761 aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc
     2821 ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc
     2881 tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac gctcaggaca
     2941 gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa ggacatttta
     3001 ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg aacaggcgag
     3061 gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg tgaacgccga
     3121 tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc gggatttggg
     3181 tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc
     3241 tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg
     3301 ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt tggggggagc
     3361 gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg
     3421 aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt cttgagccct
     3481 tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc tggggaccct
     3541 gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt
     3601 tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc
     3661 gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg
     3721 cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat
     3781 cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg
     3841 gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg
     3901 ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca
     3961 atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc gtttttggct
     4021 tttttgttag acaacatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     4081 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     4141 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     4201 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     4261 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     4321 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggagggcct cctggcgccg
     4381 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     4441 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggccgccga gcgcgccggg
     4501 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     4561 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     4621 aagcccggtg cctaggctag tatgtaagcc tagtcttaga taataaaatc gctatccatc
     4681 gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag
     4741 aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg
     4801 gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg
     4861 gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg
     4921 agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag
     4981 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5041 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt
     5101 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5161 acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5221 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5281 gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5341 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg
     5401 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5461 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5521 agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg
     5581 gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5641 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5701 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5761 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5821 gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac
     5881 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     5941 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc cgaatcgttt
     6001 aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc
     6061 gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag
     6121 cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg
     6181 gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag
     6241 gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc
     6301 atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta
     6361 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     6421 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     6481 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     6541 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     6601 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     6661 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt
     6721 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     6781 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     6841 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     6901 tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac
     6961 tcgacacacc cgccagcggc cgc
//
