LOCUS       sequence-319544-        7455 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 7455)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7455)
  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-m4134293
FEATURES             Location/Qualifiers
     source          1..7455
                     /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             1432..1497
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1566..1687
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1577..1596)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1631..1650
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1802..1848
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1878..2014
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2059..2130
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2356..2944)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2436..2455)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3444..4653
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4490..4509
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4657..5685
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5712..5760
                     /note="synthetic polyadenylation signal"
     CDS             join(6026..6094,6095..6886)
                     /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(6244..6263)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6899..7035
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7187..7258
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7263..7309)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7327..7375
                     /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 ttcagtcgat ggattataag
     1441 gaccacgatg gagactacaa agatcatgac attgattaca aggacgacga tgacaaataa
     1501 ggatccgcag gcctctgcta gcttgactga ctgagataca gcgtaccttc agctcacaga
     1561 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     1621 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     1681 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     1741 ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc
     1801 gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc
     1861 cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     1921 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     1981 atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc
     2041 tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga
     2101 agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac
     2161 ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc
     2221 tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac
     2281 gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2341 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2401 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2461 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2521 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2581 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2641 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2701 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2761 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2821 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2881 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2941 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3001 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3061 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3121 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3181 atttaaattt ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat
     3241 ccatcttgca ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct
     3301 tggctattgc ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc
     3361 gggatcaccc gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg
     3421 aatatcgaaa tcggggcgcg cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc
     3481 ccccctcgtc acggcgagcg ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct
     3541 tccgcccgga cgctcaggac agcggcccgc tgctcataag actcggcctt agaaccccag
     3601 tatcagcaga aggacatttt aggacgggac ttgggtgact ctagggcact ggttttcttt
     3661 ccagagagcg gaacaggcga ggaaaagtag tcccttctcg gcgattctgc ggagggatct
     3721 ccgtggggcg gtgaacgccg atgattatat aaggacgcgc cgggtgtggc acagctagtt
     3781 ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg
     3841 gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc cgccgggccg ctcggtggga
     3901 cggaagcgtg tggagagacc gccaagggct gtagtctggg tccgcgagca aggttgccct
     3961 gaactggggg ttggggggag cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag
     4021 acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa
     4081 gaacccaagg tcttgagccc ttcgctaatg cgggaaagct cttattcggg tgagatgggc
     4141 tgggcaccat ctggggaccc tgacgtgaag tttgtcactg actggagaac tcggtttgtc
     4201 gtctgttgcg ggggcggcag ttatggcggt gccgttgggc agtgcacccg tacctttggg
     4261 agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa tgcagggtgg
     4321 ggccacctgc cggtaggtgt gcggtaggct tttctccgtc gcaggacgca gggttcgggc
     4381 ctagggtagg ctctcctgaa tcgacaggcg ccggacctct ggtgagggga gggataagtg
     4441 aggcgtcagt ttctttggtc ggttttatgt acctatcttc ttaagtagct gaagctccgg
     4501 ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg aagtttttta ggcacctttt
     4561 gaaatgtaat catttgggtc aatatgtaat tttcagtgtt agacttgtaa attgtccgct
     4621 aaattctggc cgtttttggc ttttttgtta gacaacatgg gtaaaaagcc tgaactcacc
     4681 gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag
     4741 ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc
     4801 ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt
     4861 gcatccgccg cgctcccgat tccggaagtg cttgacattg gggagttcag cgagagcctg
     4921 acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa
     4981 ctgcccgctg ttctgcagcc ggtcgcggag gcaatggatg ccatcgctgc cgccgatctt
     5041 agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg
     5101 cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac
     5161 gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac
     5221 tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac
     5281 aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac
     5341 gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc
     5401 tacttcgagc ggaggcatcc ggagcttgca ggatcgcccc ggctccgggc gtatatgctc
     5461 cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct
     5521 tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca
     5581 caaatcgccc gcagaagcgc cgccgtctgg accgatggct gtgtagaagt actcgccgat
     5641 agtggaaacc gacgccccag cactcgtccg agggcaaagg aataagctag tatgtaagcc
     5701 tagtcttaga taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg
     5761 tgtaacgcta ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat
     5821 ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat
     5881 cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata
     5941 ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga
     6001 gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc
     6061 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     6121 agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg
     6181 taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt
     6241 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     6301 catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac
     6361 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6421 ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa
     6481 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6541 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt
     6601 aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga
     6661 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6721 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6781 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6841 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc
     6901 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca
     6961 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca
     7021 gaaatttatc cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg
     7081 tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg
     7141 tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt
     7201 agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa
     7261 ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc
     7321 gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta
     7381 actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc
     7441 atatatggag ttccg
//
