LOCUS       sequence-307273-        9980 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 9980)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 9980)
  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-m4133779
FEATURES             Location/Qualifiers
     source          1..9980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..47)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    65..113
                     /note="synthetic polyadenylation signal"
     enhancer        194..497
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        499..701
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     652..672
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4595..4615
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4616..4660
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    4713..4834
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4724..4743)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4778..4797
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4949..4995
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5025..5161
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5206..5277
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(5503..6091)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5583..5602)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6591..7800
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     7637..7656
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             7804..8403
                     /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(7804..7823)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8300..8320
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    8430..8478
                     /note="synthetic polyadenylation signal"
     CDS             join(8744..8812,8813..9604)
                     /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(8962..8981)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      9617..9753
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    9905..9976
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
ORIGIN
        1 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt
       61 tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac
      121 tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat
      181 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      241 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      301 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      361 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      421 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      481 tcatcgctat taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      541 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      601 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      661 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
      721 tcagatcttt gtcgatccta ccatccactc gacacacccg ccagcggccg ccaccatgtt
      781 tgtttttctt gttttattgc cactagtctc tagtcagtgt gttaatctta caaccagaac
      841 tcaattaccc cctgcataca ctaattcttt cacacgtggt gtttattacc ctgacaaagt
      901 tttcagatcc tcagttttac attcaactca ggacttgttc ttacctttct tttccaatgt
      961 tacttggttc catgctatac atgtctctgg gaccaatggt actaagaggt ttgataaccc
     1021 tgtcctacca tttaatgatg gtgtttattt tgcttccact gagaagtcta acataataag
     1081 aggctggatt tttggtacta ctttagattc gaagacccag tccctactta ttgttaataa
     1141 cgctactaat gttgttatta aagtctgtga atttcaattt tgtaatgatc catttttggg
     1201 tgtttattac cacaaaaaca acaaaagttg gatggaaagt gagttcagag tttattctag
     1261 tgcgaataat tgcacttttg aatatgtctc tcagcctttt cttatggacc ttgaaggaaa
     1321 acagggtaat ttcaaaaatc ttagggaatt tgtgtttaag aatattgatg gttattttaa
     1381 aatatattct aagcacacgc ctattaattt agtgcgtgat ctccctcagg gtttttcggc
     1441 tttagaacca ttggtagatt tgccaatagg tattaacatc actaggtttc aaactttact
     1501 tgctttacat agaagttatt tgactcctgg tgattcttct tcaggttgga cagctggtgc
     1561 tgcagcttat tatgtgggtt atcttcaacc taggactttt ctattaaaat ataatgaaaa
     1621 tggaaccatt acagatgctg tagactgtgc acttgaccct ctctcagaaa caaagtgtac
     1681 gttgaaatcc ttcactgtag aaaaaggaat ctatcaaact tctaacttta gagtccaacc
     1741 aacagaatct attgttagat ttcctaatat tacaaacttg tgcccttttg gtgaagtttt
     1801 taacgccacc agatttgcat ctgtttatgc ttggaacagg aagagaatca gcaactgtgt
     1861 tgctgattat tctgtcctat ataattccgc atcattttcc acttttaagt gttatggagt
     1921 gtctcctact aaattaaatg atctctgctt tactaatgtc tatgcagatt catttgtaat
     1981 tagaggtgat gaagtcagac aaatcgctcc agggcaaact ggaaagattg ctgattataa
     2041 ttataaatta ccagatgatt ttacaggctg cgttatagct tggaattcta acaatcttga
     2101 ttctaaggtt ggtggtaatt ataattacct gtatagattg tttaggaagt ctaatctcaa
     2161 accttttgag agagatattt caactgaaat ctatcaggcc ggtagcacac cttgtaatgg
     2221 tgttgaaggt tttaattgtt actttccttt acaatcatat ggtttccaac ccactaatgg
     2281 tgttggttac caaccataca gagtagtagt actttctttt gaacttctac atgcaccagc
     2341 aactgtttgt ggacctaaaa agtctactaa tttggttaaa aacaaatgtg tcaatttcaa
     2401 cttcaatggt ttaacaggca caggtgttct tactgagtct aacaaaaagt ttctgccttt
     2461 ccaacaattt ggcagagaca ttgctgacac tactgatgct gtccgtgatc cacagacact
     2521 tgagattctt gacattacac catgttcttt tggtggtgtc agtgttataa caccaggaac
     2581 aaatacttct aaccaggttg ctgttcttta tcaggatgtt aactgcacag aagtccctgt
     2641 tgctattcat gcagatcaac ttactcctac ttggcgtgtt tattctacag gttctaatgt
     2701 ttttcaaaca cgtgcaggct gtttaatagg ggctgaacat gtcaacaact catatgagtg
     2761 tgacataccc attggtgcag gtatatgcgc tagttatcag actcagacta attctcctcg
     2821 gcgggcacgt agtgtagcta gtcaatccat cattgcctac actatgtcac ttggtgcaga
     2881 aaattcagtt gcttactcta ataactctat tgccataccc acaaatttta ctattagtgt
     2941 taccacagaa attctaccag tgtctatgac caagacatca gtagattgta caatgtacat
     3001 ttgtggtgat tcaactgaat gcagcaatct tttgttgcaa tatggcagtt tttgtacaca
     3061 attaaaccgt gctttaactg gaatagctgt tgaacaagac aaaaacaccc aagaagtttt
     3121 tgcacaagtc aaacaaattt acaaaacacc accaattaaa gattttggtg gttttaattt
     3181 ttcacaaata ttaccagatc catcaaaacc aagcaagagg tcatttattg aagatctact
     3241 tttcaacaaa gtgacacttg cagatgctgg cttcatcaaa caatatggtg attgccttgg
     3301 tgatattgct gctagagacc tcatttgtgc acaaaagttt aacggcctta ctgttttgcc
     3361 acctttgctc acagatgaaa tgattgctca atacacttct gcactgttag cgggtacaat
     3421 cacttctggt tggacctttg gtgcaggtgc tgcattacaa ataccatttg ctatgcaaat
     3481 ggcttatagg tttaatggta ttggagttac acagaatgtt ctctatgaga accaaaaatt
     3541 gattgccaac caatttaata gtgctattgg caaaattcaa gactcacttt cttccacagc
     3601 aagtgcactt ggaaaacttc aagatgtggt caaccaaaat gcacaagctt taaacacgct
     3661 tgttaaacaa cttagctcca attttggtgc aatttcaagt gttttaaatg atatcctttc
     3721 acgtcttgac aaagttgagg ctgaagtgca aattgatagg ttgatcacag gcagacttca
     3781 aagtttgcag acatatgtga ctcaacaatt aattagagct gcagaaatca gagcttctgc
     3841 taatcttgct gctactaaaa tgtcagagtg tgtacttgga caatcaaaaa gagttgattt
     3901 ttgtggaaag ggctatcatc ttatgtcctt ccctcagtca gcacctcatg gtgtagtctt
     3961 cttgcatgtg acttatgtcc ctgcacaaga aaagaacttc acaactgctc ctgccatttg
     4021 tcatgatgga aaagcacact ttcctcgtga aggtgtcttt gtttcaaatg gcacacactg
     4081 gtttgtaaca caaaggaatt tttatgaacc acaaatcatt actacagaca acacatttgt
     4141 gtctggtaac tgtgatgttg taataggaat tgtcaacaac acagtttatg atcctttgca
     4201 acctgaatta gactcattca aggaggagtt agataaatat tttaagaatc atacatcacc
     4261 agatgttgat ttaggtgaca tctctggcat taatgcttca gttgtaaaca ttcaaaaaga
     4321 aattgaccgc ctcaatgagg ttgccaagaa tttaaatgaa tctctcatcg atctccaaga
     4381 acttggaaag tatgagcagt atataaaatg gccatggtac atttggctag gttttatagc
     4441 tggcttgatt gccatagtaa tggtgacaat tatgctttgc tgtatgacca gttgctgtag
     4501 ttgtctcaag ggctgttgtt cttgtggatc ctgctgcaaa tttgatgaag acgactctga
     4561 gccagtgctc aaaggagtca aattacatta cacagaaaac ctttactttc agtcgaaaga
     4621 aaccgcggca gctaagttcg agaggcagca catggacagt taagctagct tgactgactg
     4681 agatacagcg taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac
     4741 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     4801 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     4861 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     4921 tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc
     4981 ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa
     5041 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     5101 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     5161 aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt
     5221 cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc
     5281 tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag
     5341 atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc
     5401 ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca
     5461 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5521 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5581 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5641 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5701 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5761 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5821 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5881 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5941 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6001 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6061 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6121 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6181 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6241 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6301 ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc
     6361 ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc
     6421 gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta
     6481 ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct
     6541 caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggcctccgcg
     6601 ccgggttttg gcgcctcccg cgggcgcccc cctcgtcacg gcgagcgctg ccacgtcaga
     6661 cgaagggcgc aggagcgtcc tgatccttcc gcccggacgc tcaggacagc ggcccgctgc
     6721 tcataagact cggccttaga accccagtat cagcagaagg acattttagg acgggacttg
     6781 ggtgactcta gggcactggt tttctttcca gagagcggaa caggcgagga aaagtagtcc
     6841 cttctcggcg attctgcgga gggatctccg tggggcggtg aacgccgatg attatataag
     6901 gacgcgccgg gtgtggcaca gctagttccg tcgcagccgg gatttgggtc gcggttcttg
     6961 tttgtggatc gctgtgatcg tcacttggtg agtagcgggc tgctgggctg gccggggctt
     7021 tcgtggccgc cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta
     7081 gtctgggtcc gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg
     7141 cggctgttcc cgagtcttga atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa
     7201 caaggtgggg ggcatggtgg gcggcaagaa cccaaggtct tgagcccttc gctaatgcgg
     7261 gaaagctctt attcgggtga gatgggctgg gcaccatctg gggaccctga cgtgaagttt
     7321 gtcactgact ggagaactcg gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc
     7381 gttgggcagt gcacccgtac ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc
     7441 gttctgttgg cttataatgc agggtggggc cacctgccgg taggtgtgcg gtaggctttt
     7501 ctccgtcgca ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg acaggcgccg
     7561 gacctctggt gaggggaggg ataagtgagg cgtcagtttc tttggtcggt tttatgtacc
     7621 tatcttctta agtagctgaa gctccggttt tgaactatgc gctcggggtt ggcgagtgtg
     7681 ttttgtgaag ttttttaggc accttttgaa atgtaatcat ttgggtcaat atgtaatttt
     7741 cagtgttaga cttgtaaatt gtccgctaaa ttctggccgt ttttggcttt tttgttagac
     7801 aacatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     7861 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     7921 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     7981 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     8041 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     8101 ggttcccggc tggccgcgca gcaacagatg gagggcctcc tggcgccgca ccggcccaag
     8161 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     8221 ggcagcgccg tcgtgctccc cggagtggag gccgccgagc gcgccggggt gcccgccttc
     8281 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     8341 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     8401 taggctagta tgtaagccta gtcttagata ataaaatcgc tatccatcga agatggatgt
     8461 gtgttggttt tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa cgatcctctc
     8521 agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg
     8581 ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga
     8641 tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt
     8701 tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta ttcaacattt
     8761 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     8821 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg gttacatcga
     8881 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gctttccaat
     8941 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     9001 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt
     9061 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     9121 catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct
     9181 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     9241 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     9301 aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc aacagttgat
     9361 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     9421 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     9481 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     9541 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     9601 gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
     9661 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
     9721 tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa actcgactct
     9781 ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc
     9841 gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc
     9901 ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca
     9961 gcttcagcta cctctcaatt
//
