LOCUS       sequence-317395-        5021 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 5021)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5021)
  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-m4133618
FEATURES             Location/Qualifiers
     source          1..5021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(104..123)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      1669..2257
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2158..2177
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2483..2554
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2599..2735)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(2748..3539,3540..3608))
                     /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     3371..3390
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    3874..3922
                     /note="synthetic polyadenylation signal"
     CDS             complement(3949..4977)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(join(4981..5021,1..1169))
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
ORIGIN
        1 atttacaagt ctaacactga aaattacata ttgacccaaa tgattacatt tcaaaaggtg
       61 cctaaaaaac ttcacaaaac acactcgcca accccgagcg catagttcaa aaccggagct
      121 tcagctactt aagaagatag gtacataaaa ccgaccaaag aaactgacgc ctcacttatc
      181 cctcccctca ccagaggtcc ggcgcctgtc gattcaggag agcctaccct aggcccgaac
      241 cctgcgtcct gcgacggaga aaagcctacc gcacacctac cggcaggtgg ccccaccctg
      301 cattataagc caacagaacg ggtgacgtca cgacacgacg agggcgcgcg ctcccaaagg
      361 tacgggtgca ctgcccaacg gcaccgccat aactgccgcc cccgcaacag acgacaaacc
      421 gagttctcca gtcagtgaca aacttcacgt cagggtcccc agatggtgcc cagcccatct
      481 cacccgaata agagctttcc cgcattagcg aagggctcaa gaccttgggt tcttgccgcc
      541 caccatgccc cccaccttgt ttcaacgacc tcacagcccg cctcacaagc gtcttccatt
      601 caagactcgg gaacagccgc cattttgctg cgctcccccc aacccccagt tcagggcaac
      661 cttgctcgcg gacccagact acagcccttg gcggtctctc cacacgcttc cgtcccaccg
      721 agcggcccgg cggccacgaa agccccggcc agcccagcag cccgctactc accaagtgac
      781 gatcacagcg atccacaaac aagaaccgcg acccaaatcc cggctgcgac ggaactagct
      841 gtgccacacc cggcgcgtcc ttatataatc atcggcgttc accgccccac ggagatccct
      901 ccgcagaatc gccgagaagg gactactttt cctcgcctgt tccgctctct ggaaagaaaa
      961 ccagtgccct agagtcaccc aagtcccgtc ctaaaatgtc cttctgctga tactggggtt
     1021 ctaaggccga gtcttatgag cagcgggccg ctgtcctgag cgtccgggcg gaaggatcag
     1081 gacgctcctg cgcccttcgt ctgacgtggc agcgctcgcc gtgacgaggg gggcgcccgc
     1141 gggaggcgcc aaaacccggc gcggaggcca ggcgcgcccc gatttcgata ttcctcggat
     1201 ctcggataga tcgggattga ggatgtaggt tgaacttctc tcgggtgatc ccgatctcgg
     1261 atattcggga ttggagtaat acctggcgat aggcgctgcc aagcaatagc caaggcgcaa
     1321 ggccggccaa gagacttgcg atagttcgtt cgagaggtag cctgcaagat ggatcggacg
     1381 aaaggtttga agattttccg acgagggcct tggagagttc ggaaatttaa atgcaactat
     1441 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     1501 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     1561 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     1621 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     1681 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1741 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     1801 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     1861 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     1921 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     1981 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2041 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2101 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2161 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2221 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2281 acggttcctg gccttttgct ggccttttgc tcacatgatt taaatgaaac tcgtaatcgg
     2341 agaacgggtg caatagcaag gaccagatcg aacttgactg acgcaattct gagcgtagtc
     2401 gaaacccaga caagtcgatc tcgccaagta gctgacttga ctggttaatt aagtttcgat
     2461 agcccaaggt aaccaatgag attagggagg gaagaaagtt tgatgaacta cttctgaccc
     2521 ctagtggtgt ccagaaaaga ccattaaagg aatgtcttga tcgaaatcga gagatcgcct
     2581 gcaggatagc tgacgacctt aaggataaat ttctggtaag gaggacacgt atggaagtgg
     2641 gcaagttggg gaagccgtat ccgttgctga atctggcata tgtgggagta taagacgcgc
     2701 agcgtcgcat caggcatttt tttctgcgcc aatgcaccta gaatcggtta ccaatgctta
     2761 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2821 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2881 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2941 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tatcaactgt
     3001 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tacgcaaggt tgttgccatt
     3061 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3121 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3181 ggtcctccaa tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3241 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgaa
     3301 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3361 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaag
     3421 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3481 cccactcgcg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3541 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3601 atactcatgc gaaagcctcc tgctgatcct gtctcttgat agatgtttgc aaaagctagg
     3661 actcgattat acgttgaccg atcagactat caatatctag gtttaaacag atcggtacgc
     3721 tgccgtgacc aggcttgagt acaatatctg acgattggac ttcctgggtc ccaagctgga
     3781 ttccgactgg ctgactgcca agcaacgata tggatcgtcg agactcgcac tgagaggatc
     3841 gttctcggta caccaggcgc gcctagcgtt acacacacaa aaaaccaaca cacatccatc
     3901 ttcgatggat agcgatttta ttatctaaga ctaggcttac atactagctt attcctttgc
     3961 cctcggacga gtgctggggc gtcggtttcc actatcggcg agtacttcta cacagccatc
     4021 ggtccagacg gcggcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga
     4081 tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac
     4141 caagctctga tagagttggt caagaccaat gcggagcata tacgcccgga gccggggcga
     4201 tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa
     4261 ccacggcctc cagaagaaga tgttggcgac ctcgtattgg gaatccccga acatcgcctc
     4321 gctccagtca atgaccgctg ttatgcggcc attgtccgtc aggacattgt tggagccgaa
     4381 atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca tcagctcatc
     4441 gagagcctgc gcgacggacg cactgacggt gtcgtccatc acagtttgcc agtgatacac
     4501 atggggatca gcaatcgcgc atatgaaatc acgccatgta gtgtattgac cgattccttg
     4561 cggtccgaat gggccgaacc cgctcgtctg gctaagatcg gcggcagcga tggcatccat
     4621 tgcctccgcg accggctgca gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt
     4681 gacaccctgt gcacggcggg agatgcaata ggtcaggctc tcgctgaact ccccaatgtc
     4741 aagcacttcc ggaatcggga gcgcggcgga tgcaaagtgc cgataaacat aacgatcttt
     4801 gtagaaacca tcggcgcagc tatttacccg caggacatat ccacgccctc ctacatcgaa
     4861 gctgaaagca cgagattctt cgccctccga gagctgcatc aggtcggaga cgctgtcgaa
     4921 cttttcgatc agaaacttct cgacagacgt cgcggtgagt tcaggctttt tacccatgtt
     4981 gtctaacaaa aaagccaaaa acggccagaa tttagcggac a
//
