LOCUS       sequence-295054-        9970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 surface glycoprotein 
            (Spike protein).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9970)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 9970)
  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-m4134225
FEATURES             Location/Qualifiers
     source          1..9970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3824..3844
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3845..3886
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    3939..4060
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3950..3969)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4004..4023
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4175..4221
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4251..4387
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4432..4503
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4729..5317)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4809..4828)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5817..7026
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6863..6882
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             7030..7629
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7030..7049)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7526..7546
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7656..7704
                     /note="synthetic polyadenylation signal"
     CDS             join(7970..8038,8039..8830)
                     /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(8188..8207)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      8843..8979
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    9124..9195
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(9200..9246)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    9264..9312
                     /note="synthetic polyadenylation signal"
     enhancer        9393..9696
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9698..9900
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9851..9871
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgttt gtttttcttg ttttattgcc actagtctct agtcagtgtg ttaatcttac
       61 aaccagaact caattacccc ctgcatacac taattctttc acacgtggtg tttattaccc
      121 tgacaaagtt ttcagatcct cagttttaca ttcaactcag gacttgttct tacctttctt
      181 ttccaatgtt acttggttcc atgctataca tgtctctggg accaatggta ctaagaggtt
      241 tgataaccct gtcctaccat ttaatgatgg tgtttatttt gcttccactg agaagtctaa
      301 cataataaga ggctggattt ttggtactac tttagattcg aagacccagt ccctacttat
      361 tgttaataac gctactaatg ttgttattaa agtctgtgaa tttcaatttt gtaatgatcc
      421 atttttgggt gtttattacc acaaaaacaa caaaagttgg atggaaagtg agttcagagt
      481 ttattctagt gcgaataatt gcacttttga atatgtctct cagccttttc ttatggacct
      541 tgaaggaaaa cagggtaatt tcaaaaatct tagggaattt gtgtttaaga atattgatgg
      601 ttattttaaa atatattcta agcacacgcc tattaattta gtgcgtgatc tccctcaggg
      661 tttttcggct ttagaaccat tggtagattt gccaataggt attaacatca ctaggtttca
      721 aactttactt gctttacata gaagttattt gactcctggt gattcttctt caggttggac
      781 agctggtgct gcagcttatt atgtgggtta tcttcaacct aggacttttc tattaaaata
      841 taatgaaaat ggaaccatta cagatgctgt agactgtgca cttgaccctc tctcagaaac
      901 aaagtgtacg ttgaaatcct tcactgtaga aaaaggaatc tatcaaactt ctaactttag
      961 agtccaacca acagaatcta ttgttagatt tcctaatatt acaaacttgt gcccttttgg
     1021 tgaagttttt aacgccacca gatttgcatc tgtttatgct tggaacagga agagaatcag
     1081 caactgtgtt gctgattatt ctgtcctata taattccgca tcattttcca cttttaagtg
     1141 ttatggagtg tctcctacta aattaaatga tctctgcttt actaatgtct atgcagattc
     1201 atttgtaatt agaggtgatg aagtcagaca aatcgctcca gggcaaactg gaaagattgc
     1261 tgattataat tataaattac cagatgattt tacaggctgc gttatagctt ggaattctaa
     1321 caatcttgat tctaaggttg gtggtaatta taattacctg tatagattgt ttaggaagtc
     1381 taatctcaaa ccttttgaga gagatatttc aactgaaatc tatcaggccg gtagcacacc
     1441 ttgtaatggt gttgaaggtt ttaattgtta ctttccttta caatcatatg gtttccaacc
     1501 cactaatggt gttggttacc aaccatacag agtagtagta ctttcttttg aacttctaca
     1561 tgcaccagca actgtttgtg gacctaaaaa gtctactaat ttggttaaaa acaaatgtgt
     1621 caatttcaac ttcaatggtt taacaggcac aggtgttctt actgagtcta acaaaaagtt
     1681 tctgcctttc caacaatttg gcagagacat tgctgacact actgatgctg tccgtgatcc
     1741 acagacactt gagattcttg acattacacc atgttctttt ggtggtgtca gtgttataac
     1801 accaggaaca aatacttcta accaggttgc tgttctttat caggatgtta actgcacaga
     1861 agtccctgtt gctattcatg cagatcaact tactcctact tggcgtgttt attctacagg
     1921 ttctaatgtt tttcaaacac gtgcaggctg tttaataggg gctgaacatg tcaacaactc
     1981 atatgagtgt gacataccca ttggtgcagg tatatgcgct agttatcaga ctcagactaa
     2041 ttctcctcgg cgggcacgta gtgtagctag tcaatccatc attgcctaca ctatgtcact
     2101 tggtgcagaa aattcagttg cttactctaa taactctatt gccataccca caaattttac
     2161 tattagtgtt accacagaaa ttctaccagt gtctatgacc aagacatcag tagattgtac
     2221 aatgtacatt tgtggtgatt caactgaatg cagcaatctt ttgttgcaat atggcagttt
     2281 ttgtacacaa ttaaaccgtg ctttaactgg aatagctgtt gaacaagaca aaaacaccca
     2341 agaagttttt gcacaagtca aacaaattta caaaacacca ccaattaaag attttggtgg
     2401 ttttaatttt tcacaaatat taccagatcc atcaaaacca agcaagaggt catttattga
     2461 agatctactt ttcaacaaag tgacacttgc agatgctggc ttcatcaaac aatatggtga
     2521 ttgccttggt gatattgctg ctagagacct catttgtgca caaaagttta acggccttac
     2581 tgttttgcca cctttgctca cagatgaaat gattgctcaa tacacttctg cactgttagc
     2641 gggtacaatc acttctggtt ggacctttgg tgcaggtgct gcattacaaa taccatttgc
     2701 tatgcaaatg gcttataggt ttaatggtat tggagttaca cagaatgttc tctatgagaa
     2761 ccaaaaattg attgccaacc aatttaatag tgctattggc aaaattcaag actcactttc
     2821 ttccacagca agtgcacttg gaaaacttca agatgtggtc aaccaaaatg cacaagcttt
     2881 aaacacgctt gttaaacaac ttagctccaa ttttggtgca atttcaagtg ttttaaatga
     2941 tatcctttca cgtcttgaca aagttgaggc tgaagtgcaa attgataggt tgatcacagg
     3001 cagacttcaa agtttgcaga catatgtgac tcaacaatta attagagctg cagaaatcag
     3061 agcttctgct aatcttgctg ctactaaaat gtcagagtgt gtacttggac aatcaaaaag
     3121 agttgatttt tgtggaaagg gctatcatct tatgtccttc cctcagtcag cacctcatgg
     3181 tgtagtcttc ttgcatgtga cttatgtccc tgcacaagaa aagaacttca caactgctcc
     3241 tgccatttgt catgatggaa aagcacactt tcctcgtgaa ggtgtctttg tttcaaatgg
     3301 cacacactgg tttgtaacac aaaggaattt ttatgaacca caaatcatta ctacagacaa
     3361 cacatttgtg tctggtaact gtgatgttgt aataggaatt gtcaacaaca cagtttatga
     3421 tcctttgcaa cctgaattag actcattcaa ggaggagtta gataaatatt ttaagaatca
     3481 tacatcacca gatgttgatt taggtgacat ctctggcatt aatgcttcag ttgtaaacat
     3541 tcaaaaagaa attgaccgcc tcaatgaggt tgccaagaat ttaaatgaat ctctcatcga
     3601 tctccaagaa cttggaaagt atgagcagta tataaaatgg ccatggtaca tttggctagg
     3661 ttttatagct ggcttgattg ccatagtaat ggtgacaatt atgctttgct gtatgaccag
     3721 ttgctgtagt tgtctcaagg gctgttgttc ttgtggatcc tgctgcaaat ttgatgaaga
     3781 cgactctgag ccagtgctca aaggagtcaa attacattac acagaaaacc tttactttca
     3841 gtcggggaaa cctataccca atccacttct cggcttagat tctacctaag ctagcttgac
     3901 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
     3961 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     4021 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     4081 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     4141 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
     4201 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
     4261 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     4321 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     4381 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
     4441 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
     4501 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
     4561 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
     4621 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
     4681 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4741 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4801 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4861 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4921 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4981 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5041 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5101 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5161 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5221 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5281 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5341 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5401 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5461 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5521 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     5581 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     5641 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     5701 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     5761 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc
     5821 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac
     5881 gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
     5941 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
     6001 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     6061 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     6121 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     6181 ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg
     6241 gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg
     6301 gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca
     6361 aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg
     6421 ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta
     6481 atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg
     6541 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
     6601 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
     6661 tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg tgtgcggtag
     6721 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
     6781 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
     6841 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
     6901 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
     6961 aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg
     7021 ttagacaaca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     7081 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     7141 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     7201 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     7261 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     7321 ttgagcggtt cccggctggc cgcgcagcaa cagatggagg gcctcctggc gccgcaccgg
     7381 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     7441 ggtctgggca gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc cggggtgccc
     7501 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     7561 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     7621 ggtgcctagg ctagtatgta agcctagtct tagataataa aatcgctatc catcgaagat
     7681 ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat
     7741 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     7801 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     7861 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     7921 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca
     7981 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     8041 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta
     8101 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt
     8161 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     8221 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc
     8281 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     8341 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa
     8401 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     8461 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     8521 ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca
     8581 gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     8641 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     8701 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     8761 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     8821 gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     8881 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     8941 cttccatacg tgtcctcctt accagaaatt tatccttaag atcgtttaaa ctcgactctg
     9001 gctctatcga atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg
     9061 tcgggcatcg aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc
     9121 cgacatcttg gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag
     9181 cttcagctac ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt
     9241 atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat
     9301 agcgatttta ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg
     9361 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     9421 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     9481 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     9541 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     9601 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     9661 cagtacatct acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc
     9721 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     9781 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     9841 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     9901 ggtttagtga accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc
     9961 cagcggccgc
//
