LOCUS       sequence-327169-        7413 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 7413)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7413)
  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-m4134503
FEATURES             Location/Qualifiers
     source          1..7413
                     /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    1524..1645
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1535..1554)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1589..1608
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1760..1806
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1836..1972
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2017..2088
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2314..2902)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2394..2413)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3402..4611
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4448..4467
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4615..5643
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5670..5718
                     /note="synthetic polyadenylation signal"
     CDS             join(5984..6052,6053..6844)
                     /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(6202..6221)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6857..6993
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7145..7216
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7221..7267)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7285..7333
                     /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 atgcgtaagg atccgcaggc ctctgctagc ttgactgact gagatacagc
     1501 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1561 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1621 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1681 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg
     1741 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1801 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg
     1861 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     1921 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca
     1981 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg
     2041 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt
     2101 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg
     2161 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg
     2221 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc
     2281 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2341 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2401 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2461 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2521 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2581 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2641 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2701 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2761 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2821 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2881 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2941 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3001 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3061 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3121 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct
     3181 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc
     3241 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa
     3301 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga
     3361 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt
     3421 ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg
     3481 caggagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac
     3541 tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct
     3601 agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc
     3661 gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg
     3721 ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat
     3781 cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg
     3841 ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc
     3901 cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc
     3961 ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg
     4021 gggcatggtg ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg ggaaagctct
     4081 tattcgggtg agatgggctg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac
     4141 tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag
     4201 tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg
     4261 gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc
     4321 aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc ggacctctgg
     4381 tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt
     4441 aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa
     4501 gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt tcagtgttag
     4561 acttgtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga caacatgggt
     4621 aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc
     4681 gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta
     4741 ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt
     4801 tatgtttatc ggcactttgc atccgccgcg ctcccgattc cggaagtgct tgacattggg
     4861 gagttcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa
     4921 gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc aatggatgcc
     4981 atcgctgccg ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc
     5041 ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac
     5101 tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg
     5161 atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc
     5221 aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg
     5281 ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt
     5341 atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccccgg
     5401 ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc
     5461 aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc
     5521 gggactgtcg ggcgtacaca aatcgcccgc agaagcgccg ccgtctggac cgatggctgt
     5581 gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa
     5641 taagctagta tgtaagccta gtcttagata ataaaatcgc tatccatcga agatggatgt
     5701 gtgttggttt tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa cgatcctctc
     5761 agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg
     5821 ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga
     5881 tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt
     5941 tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta ttcaacattt
     6001 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6061 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg gttacatcga
     6121 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gctttccaat
     6181 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     6241 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt
     6301 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     6361 catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct
     6421 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     6481 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     6541 aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc aacagttgat
     6601 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6661 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6721 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6781 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6841 gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
     6901 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
     6961 tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa actcgactct
     7021 ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc
     7081 gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc
     7141 ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca
     7201 gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
     7261 tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga
     7321 tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg
     7381 gtcattagtt catagcccat atatggagtt ccg
//
