LOCUS       sequence-345630-        6968 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 6968)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6968)
  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-m4134517
FEATURES             Location/Qualifiers
     source          1..6968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1408..1428
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1429..1455
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1508..1629
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1519..1538)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1573..1592
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1744..1790
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1820..1956
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2001..2072
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2298..2886)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2378..2397)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3386..4595
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4432..4451
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4599..5198
                     /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(4599..4618)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5095..5115
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5225..5273
                     /note="synthetic polyadenylation signal"
     CDS             join(5539..5607,5608..6399)
                     /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(5757..5776)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6412..6548
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6700..6771
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6776..6822)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6840..6888
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatggatct gtttatgcgt
      601 atattcacca tcggcacggt tacattgaag caaggggaaa ttaaagacgc aactcccagt
      661 gacttcgtac gggcgaccgc tacaattccg atccaggcca gcctcccttt tggatggctt
      721 atcgtcggcg tggccctact ggctgtgttt cagtcggcat caaagataat cactctgaaa
      781 aagcgatggc agttagccct tagcaaaggt gtgcacttcg tatgcaattt gctgctcctg
      841 ttcgtcaccg tttattccca tctgctcctg gtggctgccg gattggaggc accattttta
      901 tacctatacg ccctcgtgta ttttctgcaa tctattaact tcgtccgcat tatcatgaga
      961 ctctggctct gttggaagtg caggtctaaa aatcccttgt tatatgatgc caactacttc
     1021 ctgtgttggc acactaactg ctatgactac tgtattccat acaattcagt tacctccagt
     1081 atagtgatca catctgggga tggtacgact tcgcctattt ccgagcatga ctatcagatc
     1141 ggcgggtaca cagaaaagtg ggagagcgga gtcaaggatt gcgtggtgct tcactcctat
     1201 ttcaccagtg actactacca gctttatagc acacaacttt caacagacac cggcgtggag
     1261 catgtgactt ttttcattta taataaaatc gttgatgaac cggaggaaca cgttcagata
     1321 cacactatcg acggcagctc tggtgtcgtc aaccctgtta tggagcccat ttacgatgaa
     1381 ccaacgacca caacatctgt acctctggaa aacctttact ttcagtcgta cccctatgac
     1441 gtaccagatt atgcgtaagc tagcttgact gactgagata cagcgtacct tcagctcaca
     1501 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     1561 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     1621 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     1681 gaggtttttt aaagcaagta aaacctctac aaatgtggta ttggcccatc tctatcggta
     1741 tcgtagcata accccttggg gcctctaaac gggtcttgag gggttttttg tgcccctcgg
     1801 gccggattgc tatctaccgg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     1861 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     1921 ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtcagctatc ctgcaggcga
     1981 tctctcgatt tcgatcaaga cattccttta atggtctttt ctggacacca ctaggggtca
     2041 gaagtagttc atcaaacttt cttccctccc taatctcatt ggttaccttg ggctatcgaa
     2101 acttaattaa ccagtcaagt cagctacttg gcgagatcga cttgtctggg tttcgactac
     2161 gctcagaatt gcgtcagtca agttcgatct ggtccttgct attgcacccg ttctccgatt
     2221 acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2281 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2341 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2401 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2461 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2521 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2581 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2641 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2701 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2761 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2821 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2881 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2941 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3001 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3061 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3121 gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg
     3181 atccatcttg caggctacct ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc
     3241 cttggctatt gcttggcagc gcctatcgcc aggtattact ccaatcccga atatccgaga
     3301 tcgggatcac ccgagagaag ttcaacctac atcctcaatc ccgatctatc cgagatccga
     3361 ggaatatcga aatcggggcg cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc
     3421 gcccccctcg tcacggcgag cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc
     3481 cttccgcccg gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc
     3541 agtatcagca gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct
     3601 ttccagagag cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat
     3661 ctccgtgggg cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag
     3721 ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact
     3781 tggtgagtag cgggctgctg ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg
     3841 gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc
     3901 ctgaactggg ggttgggggg agcgcagcaa aatggcggct gttcccgagt cttgaatgga
     3961 agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc
     4021 aagaacccaa ggtcttgagc ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg
     4081 gctgggcacc atctggggac cctgacgtga agtttgtcac tgactggaga actcggtttg
     4141 tcgtctgttg cgggggcggc agttatggcg gtgccgttgg gcagtgcacc cgtacctttg
     4201 ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat aatgcagggt
     4261 ggggccacct gccggtaggt gtgcggtagg cttttctccg tcgcaggacg cagggttcgg
     4321 gcctagggta ggctctcctg aatcgacagg cgccggacct ctggtgaggg gagggataag
     4381 tgaggcgtca gtttctttgg tcggttttat gtacctatct tcttaagtag ctgaagctcc
     4441 ggttttgaac tatgcgctcg gggttggcga gtgtgttttg tgaagttttt taggcacctt
     4501 ttgaaatgta atcatttggg tcaatatgta attttcagtg ttagacttgt aaattgtccg
     4561 ctaaattctg gccgtttttg gcttttttgt tagacaacat gaccgagtac aagcccacgg
     4621 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
     4681 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
     4741 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     4801 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     4861 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     4921 agatggaggg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     4981 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     5041 tggaggccgc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     5101 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     5161 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctaggc tagtatgtaa gcctagtctt
     5221 agataataaa atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg
     5281 ctaggcgcgc ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc
     5341 gttgcttggc agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga
     5401 tattgtactc aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag
     5461 tctgatcggt caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga
     5521 tcagcaggag gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5581 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5641 gaagatcagt tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5701 cttgagagtt ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta
     5761 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5821 tattctcaga atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc
     5881 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5941 ttacttctga caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6001 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6061 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc
     6121 gaactactta ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt
     6181 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6241 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6301 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6361 atcgctgaga taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg
     6421 cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag
     6481 caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt
     6541 atccttaaga tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc
     6601 gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta
     6661 tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca
     6721 caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa
     6781 aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac
     6841 acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg
     6901 atcgagtgta gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg
     6961 gagttccg
//
