LOCUS       sequence-297474-        9977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 S (surface 
            glycoprotein).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9977)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 9977)
  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-m4252789
FEATURES             Location/Qualifiers
     source          1..9977
                     /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    9131..9202
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(9207..9253)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    9271..9319
                     /note="synthetic polyadenylation signal"
     enhancer        9400..9703
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9705..9907
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9858..9878
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgttc gtctttcttg tgttactccc cctggtttcg tcccagtgcg tgaacttgac
       61 tacgcgaacc caactgccgc cagcatatac aaatagtttc acccgtggag tatactaccc
      121 tgataaagtg tttagatcaa gcgtactaca ctctactcag gacctgtttc tccccttctt
      181 ctccaatgtc acatggtttc atgcgatcca cgttagcggc accaacggga caaagcgctt
      241 tgacaaccct gtgctgccat tcaatgatgg tgtgtatttt gcttctaccg agaagagcaa
      301 cataattcgg ggctggatct tcggaactac cctcgacagt aaaactcagt cattgctgat
      361 tgtgaataac gccacaaatg tcgttatcaa ggtctgtgaa tttcaattct gcaacgatcc
      421 attccttggg gtgtattacc ataaaaataa taagtcctgg atggagagcg aatttagggt
      481 atactcgtct gccaacaatt gtactttcga gtatgtgtct cagcccttct taatggatct
      541 ggaaggtaaa cagggcaact ttaagaacct ccgcgagttt gtattcaaaa atatagacgg
      601 atactttaag atttattcca agcacacacc aatcaatttg gtgcgggatc tcccgcaagg
      661 gttcagcgct ctggaacctc tagtggacct ccccatcggc attaacatca caagattcca
      721 gacgctgtta gccctgcata gatcatacct tactcccggt gacagtagtt ctgggtggac
      781 cgcaggcgcg gctgcatatt acgtgggata tttgcagcct cggacgtttc tgctgaaata
      841 caacgagaac ggcaccatta cagatgccgt ggattgcgcc cttgaccccc taagcgaaac
      901 taagtgtacg cttaaaagct tcaccgtcga gaagggaatt taccagactt ccaactttag
      961 ggtccagccg acagaatcaa tagttaggtt cccaaatatc accaatttgt gcccttttgg
     1021 ggaagtcttc aatgctacac gatttgcttc ggtttatgcc tggaacagga agcgtatctc
     1081 aaattgcgtc gcggactaca gcgtgctcta taatagtgca agtttctcga cctttaaatg
     1141 ttatggcgtg tccccgacca aactaaatga cctctgtttt acaaacgttt acgccgattc
     1201 ttttgtgatt cgcggtgatg aggtacgcca aatcgcaccg ggccagactg gcaagattgc
     1261 cgattacaac tacaagttgc ctgacgactt caccggatgt gttatagcct ggaatagcaa
     1321 taatctggat tctaaggtcg gaggtaacta taactacctg tatagactgt tccgtaagtc
     1381 aaacctgaaa ccctttgagc gggacatctc cacagagatt taccaagcag ggtctacccc
     1441 ctgcaatggg gttgaaggat tcaattgcta ctttccatta cagagctacg ggttccagcc
     1501 tacgaacggg gttggttacc agccttatag ggtagtggtg ctgtcgttcg aactgttgca
     1561 cgctccagct acagtgtgcg ggcctaaaaa atcaactaac ctggttaaaa ataagtgtgt
     1621 taattttaac ttcaatggct taaccggaac tggagtgctc acggagtcaa acaagaaatt
     1681 tctaccgttt cagcaattcg ggagggatat cgcagacact acggacgcgg tcagagatcc
     1741 acagaccctt gagatccttg acataacacc ctgctctttc ggaggtgtct ccgtaattac
     1801 accaggcact aatacctcta atcaggtggc cgtgctttat caggatgtta actgtacaga
     1861 ggtcccggtg gctatccatg ccgaccaatt gacacctacg tggcgcgtct attctaccgg
     1921 ctccaacgtc ttccagaccc gggctggatg cttgataggc gcagaacacg taaacaactc
     1981 ttacgaatgt gatattccca ttggagccgg gatctgcgca agctaccaga cccaaacaaa
     2041 cagtcccaga cgggctcgta gcgtggcctc tcagagcata atagcctata ccatgtccct
     2101 tggcgcggag aatagtgtgg catactcaaa taatagcatc gctatcccca ctaactttac
     2161 tatttccgtg acgactgaga ttctgccagt gtctatgact aagacatccg tggattgcac
     2221 aatgtatatt tgtggggatt ccacagagtg tagcaacttg ttgctgcaat atggatcatt
     2281 ctgtacgcag ctgaatcgcg ccctcaccgg catcgctgtg gaacaggaca agaacactca
     2341 agaggtcttt gctcaggtga agcaaatcta caagacccca cctataaaag acttcggggg
     2401 ttttaacttc tcgcagattc tccccgaccc cagcaagcct tcaaaaagga gctttattga
     2461 ggacctgttg ttcaacaagg ttactttagc cgatgccggt tttatcaaac agtacggcga
     2521 ctgtcttggc gatatagcag cccgggatct gatttgcgca caaaaattca atggccttac
     2581 cgtactgcca ccccttctca ccgacgaaat gatcgcgcag tacacctccg cactgctggc
     2641 tgggaccatc acttcgggat ggacctttgg ggctggtgct gctttgcaaa ttcccttcgc
     2701 catgcagatg gcctatcgat tcaatggcat aggggtaaca cagaacgtat tgtatgaaaa
     2761 tcaaaagcta atcgctaacc agtttaattc agccattggt aagatccagg attcactctc
     2821 cagtacagcc tctgccctgg gaaagctcca ggacgtggtc aaccagaacg cgcaggcatt
     2881 gaatacactc gtcaaacagc tgagtagcaa tttcggcgca atcagtagtg tcttaaacga
     2941 tatactgagt aggcttgata aagtagaggc cgaggtccag attgacagac taatcacggg
     3001 gcgtcttcag agtttacaga cctatgtgac ccaacagctg atccgagcag cggaaattcg
     3061 agcaagcgcg aacctggccg ctactaaaat gagtgagtgc gtgctgggcc aaagtaaacg
     3121 agtcgacttt tgcggcaaag ggtatcatct catgtctttc cctcagtccg cccctcacgg
     3181 tgttgttttc ctccacgtga catacgttcc cgcgcaggaa aagaatttta ctacggcacc
     3241 cgctatttgc catgacggaa aggctcattt tccacgggag ggagtgttcg tttcaaacgg
     3301 gacacactgg tttgtaactc agcggaactt ttatgagcct caaatcatca cgacagacaa
     3361 taccttcgtg agtggcaact gcgatgtagt cataggcatt gtgaataaca ctgtttatga
     3421 tccacttcag ccggaactcg acagcttcaa agaggaattg gataagtact ttaaaaatca
     3481 taccagtcca gacgtagatc tcggcgatat atcgggcatt aacgccagtg tggtcaatat
     3541 acaaaaggag attgacagac tgaatgaagt agctaagaac ctcaacgaat ctctgattga
     3601 cctgcaagag ctagggaagt acgagcagta tatcaaatgg ccatggtaca tatggctggg
     3661 ttttatcgcg ggtcttattg ccatagtcat ggtgacgatt atgttgtgct gtatgacctc
     3721 ctgctgttct tgtttgaaag gctgctgctc ctgtggtagc tgttgcaaat tcgatgaaga
     3781 tgactctgaa cccgtgttga aaggcgttaa gctacactac 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 agccggtagg 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 atcccgaatc gtttaaactc
     9001 gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat
     9061 ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg
     9121 cggctcccga catcttggac cattagctcc acaggtatct tcttccctct agtggtcata
     9181 acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca
     9241 aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc
     9301 gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat
     9361 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     9421 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     9481 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     9541 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     9601 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     9661 tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca
     9721 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     9781 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     9841 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     9901 cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca
     9961 cacccgccag cggccgc
//
