LOCUS       sequence-300776-        5076 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 5076)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5076)
  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-m4134093
FEATURES             Location/Qualifiers
     source          1..5076
                     /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"
     CDS             join(3069..3137,3138..3929)
                     /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(3287..3306)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3942..4078
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4230..4301
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4306..4352)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4370..4418
                     /note="synthetic polyadenylation signal"
     enhancer        4499..4802
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4804..5006
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4957..4977
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5075..5076,1..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 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     2881 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     2941 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     3001 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     3061 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3121 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3181 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3241 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3301 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3361 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3421 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3481 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3541 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3601 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     3661 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     3721 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3781 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3841 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3901 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     3961 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     4021 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     4081 tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc
     4141 gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt
     4201 acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt
     4261 cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa
     4321 gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa
     4381 ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta
     4441 gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     4501 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     4561 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     4621 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     4681 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     4741 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     4801 tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     4861 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     4921 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     4981 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga
     5041 tcctaccatc cactcgacac acccgccagc ggccgc
//
