LOCUS       sequence-295521-        9955 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 9955)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 9955)
  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..9955
                     /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..3871
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3924..4045
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3935..3954)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3989..4008
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4160..4206
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4236..4372
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4417..4488
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4714..5302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4794..4813)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5802..7011
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6848..6867
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             7015..7614
                     /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(7015..7034)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7511..7531
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7641..7689
                     /note="synthetic polyadenylation signal"
     CDS             join(7955..8023,8024..8815)
                     /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(8173..8192)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      8828..8964
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    9109..9180
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(9185..9231)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    9249..9297
                     /note="synthetic polyadenylation signal"
     enhancer        9378..9681
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9683..9885
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9836..9856
                     /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 gtcgtacccc tatgacgtac cagattatgc gtaagctagc ttgactgact gagatacagc
     3901 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3961 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     4021 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     4081 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg
     4141 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt
     4201 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg
     4261 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     4321 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca
     4381 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg
     4441 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt
     4501 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg
     4561 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg
     4621 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc
     4681 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4741 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4801 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4861 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4921 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4981 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5041 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5101 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5161 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5221 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5281 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5341 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5401 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5461 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5521 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct
     5581 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc
     5641 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa
     5701 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga
     5761 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt
     5821 ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg
     5881 caggagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac
     5941 tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct
     6001 agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc
     6061 gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg
     6121 ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat
     6181 cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg
     6241 ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc
     6301 cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc
     6361 ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg
     6421 gggcatggtg ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg ggaaagctct
     6481 tattcgggtg agatgggctg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac
     6541 tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag
     6601 tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg
     6661 gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc
     6721 aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc ggacctctgg
     6781 tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt
     6841 aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa
     6901 gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt tcagtgttag
     6961 acttgtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga caacatgacc
     7021 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     7081 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     7141 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     7201 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     7261 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     7321 ctggccgcgc agcaacagat ggagggcctc ctggcgccgc accggcccaa ggagcccgcg
     7381 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     7441 gtcgtgctcc ccggagtgga ggccgccgag cgcgccgggg tgcccgcctt cctggagacc
     7501 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     7561 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctaggctagt
     7621 atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg tgtgttggtt
     7681 ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt
     7741 ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt gggacccagg
     7801 aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa
     7861 acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt ttgcaaacat
     7921 ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt tccgtgtcgc
     7981 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     8041 gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg aactggatct
     8101 caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa tgatgagcac
     8161 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     8221 cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag tcacagaaaa
     8281 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     8341 taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc taaccgcttt
     8401 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     8461 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caaccttgcg
     8521 taaactatta actggcgaac tacttactct agcttcccgg caacagttga tagactggat
     8581 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     8641 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     8701 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     8761 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaaccgat
     8821 tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     8881 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     8941 tccttaccag aaatttatcc ttaagatcgt ttaaactcga ctctggctct atcgaatctc
     9001 cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct
     9061 cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca
     9121 ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc
     9181 aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt
     9241 gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc
     9301 taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc
     9361 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     9421 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     9481 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     9541 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     9601 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     9661 ttagtcatcg ctattaccat gctgatgcgg ttttggcagt acatcaatgg gcgtggatag
     9721 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     9781 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     9841 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
     9901 cagatcagat ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgc
//
