LOCUS       sequence-295942-        5052 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 5052)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5052)
  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-m4133669
FEATURES             Location/Qualifiers
     source          1..5052
                     /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..868
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    921..1042
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(932..951)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     986..1005
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1157..1203
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1233..1369
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1414..1485
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1711..2299)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1791..1810)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3045..3113,3114..3905)
                     /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(3263..3282)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3918..4054
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4206..4277
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4282..4328)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4346..4394
                     /note="synthetic polyadenylation signal"
     enhancer        4475..4778
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4780..4982
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4933..4953
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5051..5052,1..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 caccatcacc atcaccatta agctagcttg actgactgag atacagcgta
      901 ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca caactagaat
      961 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     1021 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     1081 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc
     1141 atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1201 ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg
     1261 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     1321 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct
     1381 atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca
     1441 ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc
     1501 ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct
     1561 gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac
     1621 ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag
     1681 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1741 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1801 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1861 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1921 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1981 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2041 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2101 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2161 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2221 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2281 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2341 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2401 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2461 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2521 ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca
     2581 aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg
     2641 gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc
     2701 cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct
     2761 atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg tgtaccgaga acgatcctct
     2821 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     2881 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     2941 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     3001 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt
     3061 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3121 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg
     3181 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa
     3241 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     3301 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag
     3361 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3421 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc
     3481 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3541 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3601 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga
     3661 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3721 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3781 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3841 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3901 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     3961 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     4021 atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta aactcgactc
     4081 tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct
     4141 cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct
     4201 cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc
     4261 agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg
     4321 ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg
     4381 atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg
     4441 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     4501 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     4561 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     4621 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     4681 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     4741 ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt tggcagtaca
     4801 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     4861 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     4921 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     4981 ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact cgacacaccc
     5041 gccagcggcc gc
//
