LOCUS       sequence-295931-        5061 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 5061)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5061)
  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-m4133717
FEATURES             Location/Qualifiers
     source          1..5061
                     /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..877
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    930..1051
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(941..960)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     995..1014
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1166..1212
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1242..1378
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1423..1494
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1720..2308)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1800..1819)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3054..3122,3123..3914)
                     /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(3272..3291)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3927..4063
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4215..4286
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4291..4337)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4355..4403
                     /note="synthetic polyadenylation signal"
     enhancer        4484..4787
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4789..4991
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4942..4962
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5060..5061,1..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 tacccctatg acgtaccaga ttatgcgtaa gctagcttga ctgactgaga
      901 tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac
      961 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     1021 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     1081 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     1141 tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg
     1201 aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa
     1261 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     1321 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg
     1381 tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt
     1441 ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca
     1501 ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc
     1561 gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg
     1621 ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca
     1681 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1741 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1801 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1861 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1921 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1981 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2041 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2101 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2161 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2221 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2281 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2341 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2401 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2461 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2521 tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga
     2581 aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca
     2641 agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta
     2701 ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa
     2761 tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa
     2821 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     2881 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     2941 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     3001 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     3061 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3121 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     3181 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3241 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3301 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3361 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3421 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     3481 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3541 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3601 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     3661 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3721 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3781 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3841 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3901 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     3961 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     4021 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa
     4081 actcgactct ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc
     4141 tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg
     4201 atcgcggctc ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt
     4261 cataacagca gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc
     4321 cccaaggggt tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat
     4381 cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat
     4441 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     4501 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     4561 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     4621 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     4681 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     4741 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt
     4801 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     4861 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     4921 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     4981 taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc
     5041 gacacacccg ccagcggccg c
//
