LOCUS       sequence-307365-        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-m4252852
FEATURES             Location/Qualifiers
     source          1..9962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    110..181
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(186..232)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    250..298
                     /note="synthetic polyadenylation signal"
     enhancer        379..682
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        684..886
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     837..857
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4780..4800
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4801..4827
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4880..5001
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4891..4910)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4945..4964
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      5116..5162
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5192..5328
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5373..5444
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(5670..6258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5750..5769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6758..7967
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     7804..7823
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             7971..8570
                     /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(7971..7990)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8467..8487
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    8597..8645
                     /note="synthetic polyadenylation signal"
     CDS             join(8911..8979,8980..9771)
                     /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(9129..9148)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      9784..9920
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc
       61 tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc
      121 attagctcca caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct
      181 caattcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt
      241 tgcgttacac acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat
      301 ctaactgctg atcgagtgta gccagatcta gtaatcaatt acggggtcat tagttcatag
      361 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      421 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      481 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      541 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      601 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      661 attagtcatc gctattacca tgctgatgcg gttttggcag tacatcaatg ggcgtggata
      721 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      781 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      841 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
      901 tcagatcaga tctttgtcga tcctaccatc cactcgacac acccgccagc ggccgccacc
      961 atgtttgttt ttcttgtttt attgccacta gtctctagtc agtgtgttaa tcttacaacc
     1021 agaactcaat taccccctgc atacactaat tctttcacac gtggtgttta ttaccctgac
     1081 aaagttttca gatcctcagt tttacattca actcaggact tgttcttacc tttcttttcc
     1141 aatgttactt ggttccatgc tatacatgtc tctgggacca atggtactaa gaggtttgat
     1201 aaccctgtcc taccatttaa tgatggtgtt tattttgctt ccactgagaa gtctaacata
     1261 ataagaggct ggatttttgg tactacttta gattcgaaga cccagtccct acttattgtt
     1321 aataacgcta ctaatgttgt tattaaagtc tgtgaatttc aattttgtaa tgatccattt
     1381 ttgggtgttt attaccacaa aaacaacaaa agttggatgg aaagtgagtt cagagtttat
     1441 tctagtgcga ataattgcac ttttgaatat gtctctcagc cttttcttat ggaccttgaa
     1501 ggaaaacagg gtaatttcaa aaatcttagg gaatttgtgt ttaagaatat tgatggttat
     1561 tttaaaatat attctaagca cacgcctatt aatttagtgc gtgatctccc tcagggtttt
     1621 tcggctttag aaccattggt agatttgcca ataggtatta acatcactag gtttcaaact
     1681 ttacttgctt tacatagaag ttatttgact cctggtgatt cttcttcagg ttggacagct
     1741 ggtgctgcag cttattatgt gggttatctt caacctagga cttttctatt aaaatataat
     1801 gaaaatggaa ccattacaga tgctgtagac tgtgcacttg accctctctc agaaacaaag
     1861 tgtacgttga aatccttcac tgtagaaaaa ggaatctatc aaacttctaa ctttagagtc
     1921 caaccaacag aatctattgt tagatttcct aatattacaa acttgtgccc ttttggtgaa
     1981 gtttttaacg ccaccagatt tgcatctgtt tatgcttgga acaggaagag aatcagcaac
     2041 tgtgttgctg attattctgt cctatataat tccgcatcat tttccacttt taagtgttat
     2101 ggagtgtctc ctactaaatt aaatgatctc tgctttacta atgtctatgc agattcattt
     2161 gtaattagag gtgatgaagt cagacaaatc gctccagggc aaactggaaa gattgctgat
     2221 tataattata aattaccaga tgattttaca ggctgcgtta tagcttggaa ttctaacaat
     2281 cttgattcta aggttggtgg taattataat tacctgtata gattgtttag gaagtctaat
     2341 ctcaaacctt ttgagagaga tatttcaact gaaatctatc aggccggtag cacaccttgt
     2401 aatggtgttg aaggttttaa ttgttacttt cctttacaat catatggttt ccaacccact
     2461 aatggtgttg gttaccaacc atacagagta gtagtacttt cttttgaact tctacatgca
     2521 ccagcaactg tttgtggacc taaaaagtct actaatttgg ttaaaaacaa atgtgtcaat
     2581 ttcaacttca atggtttaac aggcacaggt gttcttactg agtctaacaa aaagtttctg
     2641 cctttccaac aatttggcag agacattgct gacactactg atgctgtccg tgatccacag
     2701 acacttgaga ttcttgacat tacaccatgt tcttttggtg gtgtcagtgt tataacacca
     2761 ggaacaaata cttctaacca ggttgctgtt ctttatcagg atgttaactg cacagaagtc
     2821 cctgttgcta ttcatgcaga tcaacttact cctacttggc gtgtttattc tacaggttct
     2881 aatgtttttc aaacacgtgc aggctgttta ataggggctg aacatgtcaa caactcatat
     2941 gagtgtgaca tacccattgg tgcaggtata tgcgctagtt atcagactca gactaattct
     3001 cctcggcggg cacgtagtgt agctagtcaa tccatcattg cctacactat gtcacttggt
     3061 gcagaaaatt cagttgctta ctctaataac tctattgcca tacccacaaa ttttactatt
     3121 agtgttacca cagaaattct accagtgtct atgaccaaga catcagtaga ttgtacaatg
     3181 tacatttgtg gtgattcaac tgaatgcagc aatcttttgt tgcaatatgg cagtttttgt
     3241 acacaattaa accgtgcttt aactggaata gctgttgaac aagacaaaaa cacccaagaa
     3301 gtttttgcac aagtcaaaca aatttacaaa acaccaccaa ttaaagattt tggtggtttt
     3361 aatttttcac aaatattacc agatccatca aaaccaagca agaggtcatt tattgaagat
     3421 ctacttttca acaaagtgac acttgcagat gctggcttca tcaaacaata tggtgattgc
     3481 cttggtgata ttgctgctag agacctcatt tgtgcacaaa agtttaacgg ccttactgtt
     3541 ttgccacctt tgctcacaga tgaaatgatt gctcaataca cttctgcact gttagcgggt
     3601 acaatcactt ctggttggac ctttggtgca ggtgctgcat tacaaatacc atttgctatg
     3661 caaatggctt ataggtttaa tggtattgga gttacacaga atgttctcta tgagaaccaa
     3721 aaattgattg ccaaccaatt taatagtgct attggcaaaa ttcaagactc actttcttcc
     3781 acagcaagtg cacttggaaa acttcaagat gtggtcaacc aaaatgcaca agctttaaac
     3841 acgcttgtta aacaacttag ctccaatttt ggtgcaattt caagtgtttt aaatgatatc
     3901 ctttcacgtc ttgacaaagt tgaggctgaa gtgcaaattg ataggttgat cacaggcaga
     3961 cttcaaagtt tgcagacata tgtgactcaa caattaatta gagctgcaga aatcagagct
     4021 tctgctaatc ttgctgctac taaaatgtca gagtgtgtac ttggacaatc aaaaagagtt
     4081 gatttttgtg gaaagggcta tcatcttatg tccttccctc agtcagcacc tcatggtgta
     4141 gtcttcttgc atgtgactta tgtccctgca caagaaaaga acttcacaac tgctcctgcc
     4201 atttgtcatg atggaaaagc acactttcct cgtgaaggtg tctttgtttc aaatggcaca
     4261 cactggtttg taacacaaag gaatttttat gaaccacaaa tcattactac agacaacaca
     4321 tttgtgtctg gtaactgtga tgttgtaata ggaattgtca acaacacagt ttatgatcct
     4381 ttgcaacctg aattagactc attcaaggag gagttagata aatattttaa gaatcataca
     4441 tcaccagatg ttgatttagg tgacatctct ggcattaatg cttcagttgt aaacattcaa
     4501 aaagaaattg accgcctcaa tgaggttgcc aagaatttaa atgaatctct catcgatctc
     4561 caagaacttg gaaagtatga gcagtatata aaatggccat ggtacatttg gctaggtttt
     4621 atagctggct tgattgccat agtaatggtg acaattatgc tttgctgtat gaccagttgc
     4681 tgtagttgtc tcaagggctg ttgttcttgt ggatcctgct gcaaatttga tgaagacgac
     4741 tctgagccag tgctcaaagg agtcaaatta cattacacag aaaaccttta ctttcagtcg
     4801 tacccctatg acgtaccaga ttatgcgtaa gctagcttga ctgactgaga tacagcgtac
     4861 cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     4921 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     4981 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     5041 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca
     5101 tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     5161 tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc
     5221 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     5281 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta
     5341 tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac
     5401 cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct
     5461 tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg
     5521 ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc
     5581 cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg
     5641 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5701 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5761 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5821 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5881 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5941 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6001 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6061 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6121 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6181 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6241 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6301 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6361 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6421 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6481 tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa
     6541 acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg
     6601 ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc
     6661 gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta
     6721 tccgagatcc gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg
     6781 cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca cgtcagacga agggcgcagg
     6841 agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg
     6901 ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg
     6961 cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt
     7021 ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg
     7081 tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct
     7141 gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg
     7201 gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg
     7261 agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga
     7321 gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc
     7381 atggtgggcg gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa agctcttatt
     7441 cgggtgagat gggctgggca ccatctgggg accctgacgt gaagtttgtc actgactgga
     7501 gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca
     7561 cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt
     7621 ataatgcagg gtggggccac cagccggtag gtgtgcggta ggcttttctc cgtcgcagga
     7681 cgcagggttc gggcctaggg taggctctcc tgaatcgaca ggcgccggac ctctggtgag
     7741 gggagggata agtgaggcgt cagtttcttt ggtcggtttt atgtacctat cttcttaagt
     7801 agctgaagct ccggttttga actatgcgct cggggttggc gagtgtgttt tgtgaagttt
     7861 tttaggcacc ttttgaaatg taatcatttg ggtcaatatg taattttcag tgttagactt
     7921 gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacaac atgaccgagt
     7981 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     8041 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     8101 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     8161 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     8221 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     8281 ccgcgcagca acagatggag ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     8341 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     8401 tgctccccgg agtggaggcc gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     8461 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     8521 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctag gctagtatgt
     8581 aagcctagtc ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt
     8641 gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg
     8701 acgatccata tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt
     8761 ccaatcgtca gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct
     8821 agatattgat agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat
     8881 caagagacag gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt
     8941 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     9001 gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac
     9061 agcggtaaga tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt
     9121 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     9181 cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat
     9241 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     9301 actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg
     9361 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     9421 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa
     9481 ctattaactg gcgaactact tactctagct tcccggcaac agttgataga ctggatggag
     9541 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     9601 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     9661 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     9721 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta accgattcta
     9781 ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
     9841 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
     9901 taccagaaat ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat
     9961 ct
//
