LOCUS       sequence-300352-        9962 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 9962)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 9962)
  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-m4133900
FEATURES             Location/Qualifiers
     source          1..9962
                     /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"
     CDS             4402..4422
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4423..4449
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4502..4623
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4513..4532)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4567..4586
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4738..4784
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4814..4950
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4995..5066
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(5292..5880)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5372..5391)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6380..7589
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     7426..7445
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             7593..8192
                     /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(7593..7612)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8089..8109
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    8219..8267
                     /note="synthetic polyadenylation signal"
     CDS             join(8533..8601,8602..9393)
                     /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(8751..8770)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      9406..9542
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    9694..9765
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(9770..9816)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    9834..9882
                     /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 ccatgtttgt ttttcttgtt
      601 ttattgccac tagtctctag tcagtgtgtt aatcttacaa ccagaactca attaccccct
      661 gcatacacta attctttcac acgtggtgtt tattaccctg acaaagtttt cagatcctca
      721 gttttacatt caactcagga cttgttctta cctttctttt ccaatgttac ttggttccat
      781 gctatacatg tctctgggac caatggtact aagaggtttg ataaccctgt cctaccattt
      841 aatgatggtg tttattttgc ttccactgag aagtctaaca taataagagg ctggattttt
      901 ggtactactt tagattcgaa gacccagtcc ctacttattg ttaataacgc tactaatgtt
      961 gttattaaag tctgtgaatt tcaattttgt aatgatccat ttttgggtgt ttattaccac
     1021 aaaaacaaca aaagttggat ggaaagtgag ttcagagttt attctagtgc gaataattgc
     1081 acttttgaat atgtctctca gccttttctt atggaccttg aaggaaaaca gggtaatttc
     1141 aaaaatctta gggaatttgt gtttaagaat attgatggtt attttaaaat atattctaag
     1201 cacacgccta ttaatttagt gcgtgatctc cctcagggtt tttcggcttt agaaccattg
     1261 gtagatttgc caataggtat taacatcact aggtttcaaa ctttacttgc tttacataga
     1321 agttatttga ctcctggtga ttcttcttca ggttggacag ctggtgctgc agcttattat
     1381 gtgggttatc ttcaacctag gacttttcta ttaaaatata atgaaaatgg aaccattaca
     1441 gatgctgtag actgtgcact tgaccctctc tcagaaacaa agtgtacgtt gaaatccttc
     1501 actgtagaaa aaggaatcta tcaaacttct aactttagag tccaaccaac agaatctatt
     1561 gttagatttc ctaatattac aaacttgtgc ccttttggtg aagtttttaa cgccaccaga
     1621 tttgcatctg tttatgcttg gaacaggaag agaatcagca actgtgttgc tgattattct
     1681 gtcctatata attccgcatc attttccact tttaagtgtt atggagtgtc tcctactaaa
     1741 ttaaatgatc tctgctttac taatgtctat gcagattcat ttgtaattag aggtgatgaa
     1801 gtcagacaaa tcgctccagg gcaaactgga aagattgctg attataatta taaattacca
     1861 gatgatttta caggctgcgt tatagcttgg aattctaaca atcttgattc taaggttggt
     1921 ggtaattata attacctgta tagattgttt aggaagtcta atctcaaacc ttttgagaga
     1981 gatatttcaa ctgaaatcta tcaggccggt agcacacctt gtaatggtgt tgaaggtttt
     2041 aattgttact ttcctttaca atcatatggt ttccaaccca ctaatggtgt tggttaccaa
     2101 ccatacagag tagtagtact ttcttttgaa cttctacatg caccagcaac tgtttgtgga
     2161 cctaaaaagt ctactaattt ggttaaaaac aaatgtgtca atttcaactt caatggttta
     2221 acaggcacag gtgttcttac tgagtctaac aaaaagtttc tgcctttcca acaatttggc
     2281 agagacattg ctgacactac tgatgctgtc cgtgatccac agacacttga gattcttgac
     2341 attacaccat gttcttttgg tggtgtcagt gttataacac caggaacaaa tacttctaac
     2401 caggttgctg ttctttatca ggatgttaac tgcacagaag tccctgttgc tattcatgca
     2461 gatcaactta ctcctacttg gcgtgtttat tctacaggtt ctaatgtttt tcaaacacgt
     2521 gcaggctgtt taataggggc tgaacatgtc aacaactcat atgagtgtga catacccatt
     2581 ggtgcaggta tatgcgctag ttatcagact cagactaatt ctcctcggcg ggcacgtagt
     2641 gtagctagtc aatccatcat tgcctacact atgtcacttg gtgcagaaaa ttcagttgct
     2701 tactctaata actctattgc catacccaca aattttacta ttagtgttac cacagaaatt
     2761 ctaccagtgt ctatgaccaa gacatcagta gattgtacaa tgtacatttg tggtgattca
     2821 actgaatgca gcaatctttt gttgcaatat ggcagttttt gtacacaatt aaaccgtgct
     2881 ttaactggaa tagctgttga acaagacaaa aacacccaag aagtttttgc acaagtcaaa
     2941 caaatttaca aaacaccacc aattaaagat tttggtggtt ttaatttttc acaaatatta
     3001 ccagatccat caaaaccaag caagaggtca tttattgaag atctactttt caacaaagtg
     3061 acacttgcag atgctggctt catcaaacaa tatggtgatt gccttggtga tattgctgct
     3121 agagacctca tttgtgcaca aaagtttaac ggccttactg ttttgccacc tttgctcaca
     3181 gatgaaatga ttgctcaata cacttctgca ctgttagcgg gtacaatcac ttctggttgg
     3241 acctttggtg caggtgctgc attacaaata ccatttgcta tgcaaatggc ttataggttt
     3301 aatggtattg gagttacaca gaatgttctc tatgagaacc aaaaattgat tgccaaccaa
     3361 tttaatagtg ctattggcaa aattcaagac tcactttctt ccacagcaag tgcacttgga
     3421 aaacttcaag atgtggtcaa ccaaaatgca caagctttaa acacgcttgt taaacaactt
     3481 agctccaatt ttggtgcaat ttcaagtgtt ttaaatgata tcctttcacg tcttgacaaa
     3541 gttgaggctg aagtgcaaat tgataggttg atcacaggca gacttcaaag tttgcagaca
     3601 tatgtgactc aacaattaat tagagctgca gaaatcagag cttctgctaa tcttgctgct
     3661 actaaaatgt cagagtgtgt acttggacaa tcaaaaagag ttgatttttg tggaaagggc
     3721 tatcatctta tgtccttccc tcagtcagca cctcatggtg tagtcttctt gcatgtgact
     3781 tatgtccctg cacaagaaaa gaacttcaca actgctcctg ccatttgtca tgatggaaaa
     3841 gcacactttc ctcgtgaagg tgtctttgtt tcaaatggca cacactggtt tgtaacacaa
     3901 aggaattttt atgaaccaca aatcattact acagacaaca catttgtgtc tggtaactgt
     3961 gatgttgtaa taggaattgt caacaacaca gtttatgatc ctttgcaacc tgaattagac
     4021 tcattcaagg aggagttaga taaatatttt aagaatcata catcaccaga tgttgattta
     4081 ggtgacatct ctggcattaa tgcttcagtt gtaaacattc aaaaagaaat tgaccgcctc
     4141 aatgaggttg ccaagaattt aaatgaatct ctcatcgatc tccaagaact tggaaagtat
     4201 gagcagtata taaaatggcc atggtacatt tggctaggtt ttatagctgg cttgattgcc
     4261 atagtaatgg tgacaattat gctttgctgt atgaccagtt gctgtagttg tctcaagggc
     4321 tgttgttctt gtggatcctg ctgcaaattt gatgaagacg actctgagcc agtgctcaaa
     4381 ggagtcaaat tacattacac agaaaacctt tactttcagt cgtaccccta tgacgtacca
     4441 gattatgcgt aagctagctt gactgactga gatacagcgt accttcagct cacagacatg
     4501 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     4561 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     4621 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     4681 ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc ggtatcgtag
     4741 cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc tcgggccgga
     4801 ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata
     4861 ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc acttccatac
     4921 gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc
     4981 gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg gtcagaagta
     5041 gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat cgaaacttaa
     5101 ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga ctacgctcag
     5161 aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc gattacgagt
     5221 ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5281 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5341 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5401 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5461 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5521 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5581 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5641 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5701 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5761 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5821 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5881 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5941 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6001 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6061 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt
     6121 aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat
     6181 cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc
     6241 tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga
     6301 tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata
     6361 tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc
     6421 ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct gatccttccg
     6481 cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc
     6541 agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag
     6601 agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt
     6661 ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt
     6721 cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga
     6781 gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga
     6841 agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac
     6901 tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc
     6961 ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac
     7021 ccaaggtctt gagcccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg
     7081 caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct
     7141 gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg
     7201 cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc
     7261 acctgccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag
     7321 ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc
     7381 gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt
     7441 gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa
     7501 tgtaatcatt tgggtcaata tgtaattttc agtgttagac ttgtaaattg tccgctaaat
     7561 tctggccgtt tttggctttt ttgttagaca acatgaccga gtacaagccc acggtgcgcc
     7621 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     7681 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     7741 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     7801 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     7861 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     7921 agggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     7981 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     8041 ccgccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     8101 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     8161 cctggtgcat gacccgcaag cccggtgcct aggctagtat gtaagcctag tcttagataa
     8221 taaaatcgct atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt aacgctaggc
     8281 gcgcctggtg taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct
     8341 tggcagtcag ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt
     8401 actcaagcct ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat
     8461 cggtcaacgt ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca
     8521 ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     8581 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     8641 cagttgggtg cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     8701 agttttcgcc ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc
     8761 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     8821 cagaatgact tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca
     8881 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     8941 ctgacaacga ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     9001 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     9061 gacaccacga tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta
     9121 cttactctag cttcccggca acagttgata gactggatgg aggcggataa agttgcagga
     9181 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     9241 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     9301 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     9361 gagataggtg cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa
     9421 aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg
     9481 atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt
     9541 aagatcccga atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt
     9601 acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca
     9661 gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta
     9721 tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc
     9781 tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa
     9841 aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag
     9901 tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     9961 cg
//
