LOCUS       sequence-295910-        6979 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 6979)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6979)
  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-m4134269
FEATURES             Location/Qualifiers
     source          1..6979
                     /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..895
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    948..1069
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(959..978)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1013..1032
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1184..1230
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1260..1396
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1441..1512
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1738..2326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1818..1837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2826..4035
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3872..3891
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4039..4638
                     /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(4039..4058)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4535..4555
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4665..4713
                     /note="synthetic polyadenylation signal"
     CDS             join(4979..5047,5048..5839)
                     /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(5197..5216)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5852..5988
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6133..6204
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6209..6255)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6273..6321
                     /note="synthetic polyadenylation signal"
     enhancer        6402..6705
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6707..6909
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6860..6880
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6978..6979,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggat ctgtttatgc gtatattcac catcggcacg gttacattga agcaagggga
       61 aattaaagac gcaactccca gtgacttcgt acgggcgacc gctacaattc cgatccaggc
      121 cagcctccct tttggatggc ttatcgtcgg cgtggcccta ctggctgtgt ttcagtcggc
      181 atcaaagata atcactctga aaaagcgatg gcagttagcc cttagcaaag gtgtgcactt
      241 cgtatgcaat ttgctgctcc tgttcgtcac cgtttattcc catctgctcc tggtggctgc
      301 cggattggag gcaccatttt tatacctata cgccctcgtg tattttctgc aatctattaa
      361 cttcgtccgc attatcatga gactctggct ctgttggaag tgcaggtcta aaaatccctt
      421 gttatatgat gccaactact tcctgtgttg gcacactaac tgctatgact actgtattcc
      481 atacaattca gttacctcca gtatagtgat cacatctggg gatggtacga cttcgcctat
      541 ttccgagcat gactatcaga tcggcgggta cacagaaaag tgggagagcg gagtcaagga
      601 ttgcgtggtg cttcactcct atttcaccag tgactactac cagctttata gcacacaact
      661 ttcaacagac accggcgtgg agcatgtgac ttttttcatt tataataaaa tcgttgatga
      721 accggaggaa cacgttcaga tacacactat cgacggcagc tctggtgtcg tcaaccctgt
      781 tatggagccc atttacgatg aaccaacgac cacaacatct gtacctctgg aaaaccttta
      841 ctttcagtcg aaagaaaccg cggcagctaa gttcgagagg cagcacatgg acagttaagc
      901 tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga
      961 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     1021 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     1081 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     1141 aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg
     1201 gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg
     1261 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     1321 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     1381 agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga
     1441 cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt
     1501 cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt
     1561 cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca
     1621 agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat
     1681 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1741 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1801 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1861 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1921 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1981 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2041 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2101 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2161 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2221 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2281 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2341 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2401 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2461 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2521 acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc
     2581 tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct
     2641 ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc
     2701 gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag
     2761 ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg
     2821 cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag
     2881 cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg
     2941 acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt
     3001 ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc
     3061 gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc
     3121 cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt
     3181 gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag cgggctgctg
     3241 ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg tgtggagaga
     3301 ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg ggttgggggg
     3361 agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt gaggcgggct
     3421 gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa ggtcttgagc
     3481 ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctgggcacc atctggggac
     3541 cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg cgggggcggc
     3601 agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt
     3661 gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacct gccggtaggt
     3721 gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta ggctctcctg
     3781 aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca gtttctttgg
     3841 tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg
     3901 gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta atcatttggg
     3961 tcaatatgta attttcagtg ttagacttgt aaattgtccg ctaaattctg gccgtttttg
     4021 gcttttttgt tagacaacat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     4081 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     4141 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     4201 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     4261 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     4321 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaggg cctcctggcg
     4381 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     4441 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggccgc cgagcgcgcc
     4501 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     4561 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     4621 cgcaagcccg gtgcctaggc tagtatgtaa gcctagtctt agataataaa atcgctatcc
     4681 atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc
     4741 gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag
     4801 tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc
     4861 acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa
     4921 tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat
     4981 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5041 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg
     5101 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5161 agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5221 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5281 tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5341 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg
     5401 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5461 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5521 tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc
     5581 ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5641 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5701 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5761 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5821 actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc
     5881 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     5941 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga tcgtttaaac
     6001 tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc
     6061 atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat
     6121 cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca
     6181 taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc
     6241 caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct
     6301 tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat ctagtaatca
     6361 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     6421 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     6481 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     6541 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     6601 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     6661 cctacttggc agtacatcta cgtattagtc atcgctatta ccatgctgat gcggttttgg
     6721 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     6781 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     6841 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     6901 agcagagctg gtttagtgaa ccgtcagatc agatctttgt cgatcctacc atccactcga
     6961 cacacccgcc agcggccgc
//
