LOCUS       sequence-332019-        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-m4134477
FEATURES             Location/Qualifiers
     source          1..5076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4..23)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     58..77
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(141..182)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(183..203)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     regulatory      1025..1034
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1103..1305)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1132..1152)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1307..1610
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    1691..1739
                     /note="synthetic polyadenylation signal"
     terminator      1757..1803
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    1808..1879
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2031..2167)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(2180..2971,2972..3040))
                     /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     2803..2822
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3786..4374
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4275..4294
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4600..4671
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4716..4852)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(4882..4928)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    join(5043..5076,1..88)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
       61 gtttgtccaa actcatcaat gtatcttatc atgtctgtga gctgaaggta cgctgtatct
      121 cagtcagtca agctagctta ggtagaatct aagccgagaa gtggattggg tataggtttc
      181 cccgactgaa agtaaaggtt ttccagaggt acagatgttg tggtcgttgg ttcatcgtaa
      241 atgggctcca taacagggtt gacgacacca gagctgccgt cgatagtgtg tatctgaacg
      301 tgttcctccg gttcatcaac gattttatta taaatgaaaa aagtcacatg ctccacgccg
      361 gtgtctgttg aaagttgtgt gctataaagc tggtagtagt cactggtgaa ataggagtga
      421 agcaccacgc aatccttgac tccgctctcc cacttttctg tgtacccgcc gatctgatag
      481 tcatgctcgg aaataggcga agtcgtacca tccccagatg tgatcactat actggaggta
      541 actgaattgt atggaataca gtagtcatag cagttagtgt gccaacacag gaagtagttg
      601 gcatcatata acaagggatt tttagacctg cacttccaac agagccagag tctcatgata
      661 atgcggacga agttaataga ttgcagaaaa tacacgaggg cgtataggta taaaaatggt
      721 gcctccaatc cggcagccac caggagcaga tgggaataaa cggtgacgaa caggagcagc
      781 aaattgcata cgaagtgcac acctttgcta agggctaact gccatcgctt tttcagagtg
      841 attatctttg atgccgactg aaacacagcc agtagggcca cgccgacgat aagccatcca
      901 aaagggaggc tggcctggat cggaattgta gcggtcgccc gtacgaagtc actgggagtt
      961 gcgtctttaa tttccccttg cttcaatgta accgtgccga tggtgaatat acgcataaac
     1021 agatccatgg tggcggccgc tggcgggtgt gtcgagtgga tggtaggatc gacaaagatc
     1081 tgatctgacg gttcactaaa ccagctctgc ttatatagac ctcccaccgt acacgcctac
     1141 cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt
     1201 ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt
     1261 caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcagcatg gtaatagcga
     1321 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat
     1381 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
     1441 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat
     1501 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg
     1561 ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca
     1621 tatatgggct atgaactaat gaccccgtaa ttgattacta gatctggcta cactcgatca
     1681 gcagttagat aataaaatcg ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt
     1741 gtaacgcaac gattgatagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1801 ttgaattgag aggtagctga agctgctgtt atgaccacta gagggaagaa gatacctgtg
     1861 gagctaatgg tccaagatgt cgggagccgc gatcgctgcc aaaaaggtaa gctgacgata
     1921 gctgacgaga ttcgatgccc gacgagcaaa tgagcgccac ggctgttcgc gtaagctcga
     1981 aacgacggag attcgataga gccagagtcg agtttaaacg attcgggatc ttaaggataa
     2041 atttctggta aggaggacac gtatggaagt gggcaagttg gggaagccgt atccgttgct
     2101 gaatctggca tatgtgggag tataagacgc gcagcgtcgc atcaggcatt tttttctgcg
     2161 ccaatgcacc tagaatcggt taccaatgct taatcagtga ggcacctatc tcagcgatct
     2221 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     2281 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     2341 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     2401 ctttatccgc ctccatccag tctatcaact gttgccggga agctagagta agtagttcgc
     2461 cagttaatag tttacgcaag gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     2521 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     2581 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc aatcgttgtc agaagtaagt
     2641 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     2701 catccgtaag atgcttttct gtgactggtg aatactcaac caagtcattc tgagaatagt
     2761 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     2821 gcagaacttt aaaagtgctc atcattggaa agcgttcttc ggggcgaaaa ctctcaagga
     2881 tcttaccgct gttgagatcc agttcgatgt aacccactcg cgcacccaac tgatcttcag
     2941 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     3001 aaaagggaat aagggcgaca cggaaatgtt gaatactcat gcgaaagcct cctgctgatc
     3061 ctgtctcttg atagatgttt gcaaaagcta ggactcgatt atacgttgac cgatcagact
     3121 atcaatatct aggtttaaac agatcggtac gctgccgtga ccaggcttga gtacaatatc
     3181 tgacgattgg acttcctggg tcccaagctg gattccgact ggctgactgc caagcaacga
     3241 tatggatcgt cgagactcgc actgagagga tcgttctcgg tacaccaggc gcgccccgat
     3301 ttcgatattc ctcggatctc ggatagatcg ggattgagga tgtaggttga acttctctcg
     3361 ggtgatcccg atctcggata ttcgggattg gagtaatacc tggcgatagg cgctgccaag
     3421 caatagccaa ggcgcaaggc cggccaagag acttgcgata gttcgttcga gaggtagcct
     3481 gcaagatgga tcggacgaaa ggtttgaaga ttttccgacg agggccttgg agagttcgga
     3541 aatttaaatg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3601 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     3661 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3721 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3781 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3841 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3901 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3961 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4021 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4081 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4141 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4201 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4261 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4321 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4381 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgatttaa
     4441 atgaaactcg taatcggaga acgggtgcaa tagcaaggac cagatcgaac ttgactgacg
     4501 caattctgag cgtagtcgaa acccagacaa gtcgatctcg ccaagtagct gacttgactg
     4561 gttaattaag tttcgatagc ccaaggtaac caatgagatt agggagggaa gaaagtttga
     4621 tgaactactt ctgaccccta gtggtgtcca gaaaagacca ttaaaggaat gtcttgatcg
     4681 aaatcgagag atcgcctgca ggatagctga cgaccttaag gataaatttc tggtaaggag
     4741 gacacgtatg gaagtgggca agttggggaa gccgtatccg ttgctgaatc tggcatatgt
     4801 gggagtataa gacgcgcagc gtcgcatcag gcattttttt ctgcgccaat gccggtagat
     4861 agcaatccgg cccgaggggc acaaaaaacc cctcaagacc cgtttagagg ccccaagggg
     4921 ttatgctacg ataccgatag agatgggcca ataccacatt tgtagaggtt ttacttgctt
     4981 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     5041 ttaacttgtt tattgcagct tataatggtt acaaat
//
