LOCUS       sequence-319578-        7428 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 7428)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7428)
  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-m4134431
FEATURES             Location/Qualifiers
     source          1..7428
                     /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..1470
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1539..1660
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1550..1569)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1604..1623
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1775..1821
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1851..1987
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2032..2103
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2329..2917)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2409..2428)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3417..4626
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4463..4482
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4630..5658
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5685..5733
                     /note="synthetic polyadenylation signal"
     CDS             join(5999..6067,6068..6859)
                     /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(6217..6236)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6872..7008
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7160..7231
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7236..7282)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7300..7348
                     /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 ttcagtcggg gaaacctata
     1441 cccaatccac ttctcggctt agattctacc taaggatccg caggcctctg ctagcttgac
     1501 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
     1561 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     1621 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     1681 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     1741 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
     1801 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
     1861 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     1921 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     1981 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
     2041 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
     2101 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
     2161 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
     2221 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
     2281 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2341 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2401 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2461 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2521 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2581 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2641 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2701 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2761 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2821 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2881 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2941 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3001 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3061 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3121 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     3181 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     3241 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     3301 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     3361 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc
     3421 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac
     3481 gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
     3541 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
     3601 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     3661 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     3721 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     3781 ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg
     3841 gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg
     3901 gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca
     3961 aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg
     4021 ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta
     4081 atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg
     4141 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
     4201 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
     4261 tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg tgtgcggtag
     4321 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
     4381 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
     4441 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
     4501 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
     4561 aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg
     4621 ttagacaaca tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc
     4681 gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct
     4741 ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt
     4801 ttctacaaag atcgttatgt ttatcggcac tttgcatccg ccgcgctccc gattccggaa
     4861 gtgcttgaca ttggggagtt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag
     4921 ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg
     4981 gaggcaatgg atgccatcgc tgccgccgat cttagccaga cgagcgggtt cggcccattc
     5041 ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat
     5101 ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag
     5161 gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac
     5221 gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac
     5281 tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg
     5341 ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt
     5401 gcaggatcgc cccggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag
     5461 agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc
     5521 gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgccgccgtc
     5581 tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt
     5641 ccgagggcaa aggaataagc tagtatgtaa gcctagtctt agataataaa atcgctatcc
     5701 atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc
     5761 gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag
     5821 tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc
     5881 acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa
     5941 tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat
     6001 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6061 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg
     6121 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6181 agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6241 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6301 tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6361 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg
     6421 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6481 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6541 tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc
     6601 ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6661 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6721 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6781 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6841 actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc
     6901 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     6961 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga tcccgaatcg
     7021 tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt
     7081 ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg
     7141 cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta
     7201 gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta
     7261 gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca
     7321 tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta
     7381 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccg
//
