LOCUS       sequence-300236-       10004 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 10004)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 10004)
  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-m4133141
FEATURES             Location/Qualifiers
     source          1..10004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    92..163
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(168..214)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    232..280
                     /note="synthetic polyadenylation signal"
     enhancer        361..664
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        666..868
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     819..839
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4762..4782
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4786..4851
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    4904..5025
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4915..4934)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4969..4988
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      5140..5186
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5216..5352
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5397..5468
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(5694..6282)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5774..5793)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6782..7991
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     7828..7847
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             7995..8594
                     /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(7995..8014)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8491..8511
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    8621..8669
                     /note="synthetic polyadenylation signal"
     CDS             join(8935..9003,9004..9795)
                     /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(9153..9172)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      9808..9944
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc
       61 ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc
      121 ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc
      181 aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa
      241 aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg
      301 tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      361 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      421 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      481 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      541 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      601 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      661 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      721 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      781 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      841 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      901 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgtttgt ttttcttgtt
      961 ttattgccac tagtctctag tcagtgtgtt aatcttacaa ccagaactca attaccccct
     1021 gcatacacta attctttcac acgtggtgtt tattaccctg acaaagtttt cagatcctca
     1081 gttttacatt caactcagga cttgttctta cctttctttt ccaatgttac ttggttccat
     1141 gctatacatg tctctgggac caatggtact aagaggtttg ataaccctgt cctaccattt
     1201 aatgatggtg tttattttgc ttccactgag aagtctaaca taataagagg ctggattttt
     1261 ggtactactt tagattcgaa gacccagtcc ctacttattg ttaataacgc tactaatgtt
     1321 gttattaaag tctgtgaatt tcaattttgt aatgatccat ttttgggtgt ttattaccac
     1381 aaaaacaaca aaagttggat ggaaagtgag ttcagagttt attctagtgc gaataattgc
     1441 acttttgaat atgtctctca gccttttctt atggaccttg aaggaaaaca gggtaatttc
     1501 aaaaatctta gggaatttgt gtttaagaat attgatggtt attttaaaat atattctaag
     1561 cacacgccta ttaatttagt gcgtgatctc cctcagggtt tttcggcttt agaaccattg
     1621 gtagatttgc caataggtat taacatcact aggtttcaaa ctttacttgc tttacataga
     1681 agttatttga ctcctggtga ttcttcttca ggttggacag ctggtgctgc agcttattat
     1741 gtgggttatc ttcaacctag gacttttcta ttaaaatata atgaaaatgg aaccattaca
     1801 gatgctgtag actgtgcact tgaccctctc tcagaaacaa agtgtacgtt gaaatccttc
     1861 actgtagaaa aaggaatcta tcaaacttct aactttagag tccaaccaac agaatctatt
     1921 gttagatttc ctaatattac aaacttgtgc ccttttggtg aagtttttaa cgccaccaga
     1981 tttgcatctg tttatgcttg gaacaggaag agaatcagca actgtgttgc tgattattct
     2041 gtcctatata attccgcatc attttccact tttaagtgtt atggagtgtc tcctactaaa
     2101 ttaaatgatc tctgctttac taatgtctat gcagattcat ttgtaattag aggtgatgaa
     2161 gtcagacaaa tcgctccagg gcaaactgga aagattgctg attataatta taaattacca
     2221 gatgatttta caggctgcgt tatagcttgg aattctaaca atcttgattc taaggttggt
     2281 ggtaattata attacctgta tagattgttt aggaagtcta atctcaaacc ttttgagaga
     2341 gatatttcaa ctgaaatcta tcaggccggt agcacacctt gtaatggtgt tgaaggtttt
     2401 aattgttact ttcctttaca atcatatggt ttccaaccca ctaatggtgt tggttaccaa
     2461 ccatacagag tagtagtact ttcttttgaa cttctacatg caccagcaac tgtttgtgga
     2521 cctaaaaagt ctactaattt ggttaaaaac aaatgtgtca atttcaactt caatggttta
     2581 acaggcacag gtgttcttac tgagtctaac aaaaagtttc tgcctttcca acaatttggc
     2641 agagacattg ctgacactac tgatgctgtc cgtgatccac agacacttga gattcttgac
     2701 attacaccat gttcttttgg tggtgtcagt gttataacac caggaacaaa tacttctaac
     2761 caggttgctg ttctttatca ggatgttaac tgcacagaag tccctgttgc tattcatgca
     2821 gatcaactta ctcctacttg gcgtgtttat tctacaggtt ctaatgtttt tcaaacacgt
     2881 gcaggctgtt taataggggc tgaacatgtc aacaactcat atgagtgtga catacccatt
     2941 ggtgcaggta tatgcgctag ttatcagact cagactaatt ctcctcggcg ggcacgtagt
     3001 gtagctagtc aatccatcat tgcctacact atgtcacttg gtgcagaaaa ttcagttgct
     3061 tactctaata actctattgc catacccaca aattttacta ttagtgttac cacagaaatt
     3121 ctaccagtgt ctatgaccaa gacatcagta gattgtacaa tgtacatttg tggtgattca
     3181 actgaatgca gcaatctttt gttgcaatat ggcagttttt gtacacaatt aaaccgtgct
     3241 ttaactggaa tagctgttga acaagacaaa aacacccaag aagtttttgc acaagtcaaa
     3301 caaatttaca aaacaccacc aattaaagat tttggtggtt ttaatttttc acaaatatta
     3361 ccagatccat caaaaccaag caagaggtca tttattgaag atctactttt caacaaagtg
     3421 acacttgcag atgctggctt catcaaacaa tatggtgatt gccttggtga tattgctgct
     3481 agagacctca tttgtgcaca aaagtttaac ggccttactg ttttgccacc tttgctcaca
     3541 gatgaaatga ttgctcaata cacttctgca ctgttagcgg gtacaatcac ttctggttgg
     3601 acctttggtg caggtgctgc attacaaata ccatttgcta tgcaaatggc ttataggttt
     3661 aatggtattg gagttacaca gaatgttctc tatgagaacc aaaaattgat tgccaaccaa
     3721 tttaatagtg ctattggcaa aattcaagac tcactttctt ccacagcaag tgcacttgga
     3781 aaacttcaag atgtggtcaa ccaaaatgca caagctttaa acacgcttgt taaacaactt
     3841 agctccaatt ttggtgcaat ttcaagtgtt ttaaatgata tcctttcacg tcttgacaaa
     3901 gttgaggctg aagtgcaaat tgataggttg atcacaggca gacttcaaag tttgcagaca
     3961 tatgtgactc aacaattaat tagagctgca gaaatcagag cttctgctaa tcttgctgct
     4021 actaaaatgt cagagtgtgt acttggacaa tcaaaaagag ttgatttttg tggaaagggc
     4081 tatcatctta tgtccttccc tcagtcagca cctcatggtg tagtcttctt gcatgtgact
     4141 tatgtccctg cacaagaaaa gaacttcaca actgctcctg ccatttgtca tgatggaaaa
     4201 gcacactttc ctcgtgaagg tgtctttgtt tcaaatggca cacactggtt tgtaacacaa
     4261 aggaattttt atgaaccaca aatcattact acagacaaca catttgtgtc tggtaactgt
     4321 gatgttgtaa taggaattgt caacaacaca gtttatgatc ctttgcaacc tgaattagac
     4381 tcattcaagg aggagttaga taaatatttt aagaatcata catcaccaga tgttgattta
     4441 ggtgacatct ctggcattaa tgcttcagtt gtaaacattc aaaaagaaat tgaccgcctc
     4501 aatgaggttg ccaagaattt aaatgaatct ctcatcgatc tccaagaact tggaaagtat
     4561 gagcagtata taaaatggcc atggtacatt tggctaggtt ttatagctgg cttgattgcc
     4621 atagtaatgg tgacaattat gctttgctgt atgaccagtt gctgtagttg tctcaagggc
     4681 tgttgttctt gtggatcctg ctgcaaattt gatgaagacg actctgagcc agtgctcaaa
     4741 ggagtcaaat tacattacac agaaaacctt tactttcagt cgatggatta taaggaccac
     4801 gatggagact acaaagatca tgacattgat tacaaggacg acgatgacaa ataagctagc
     4861 ttgactgact gagatacagc gtaccttcag ctcacagaca tgataagata cattgatgag
     4921 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     4981 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     5041 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     5101 ctctacaaat gtggtattgg cccatctcta tcggtatcgt agcataaccc cttggggcct
     5161 ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg gattgctatc taccggcatt
     5221 ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat
     5281 tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa
     5341 atttatcctt aaggtcgtca gctatcctgc aggcgatctc tcgatttcga tcaagacatt
     5401 cctttaatgg tcttttctgg acaccactag gggtcagaag tagttcatca aactttcttc
     5461 cctccctaat ctcattggtt accttgggct atcgaaactt aattaaccag tcaagtcagc
     5521 tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt cagtcaagtt
     5581 cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta aatcatgtga
     5641 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5701 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5761 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5821 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5881 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5941 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6001 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6061 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6121 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6181 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6241 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6301 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6361 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6421 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6481 atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc gaactctcca
     6541 aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg
     6601 aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt ggcagcgcct
     6661 atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga gagaagttca
     6721 acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc
     6781 tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct
     6841 gccacgtcag acgaagggcg caggagcgtc ctgatccttc cgcccggacg ctcaggacag
     6901 cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag
     6961 gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg
     7021 aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat
     7081 gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt
     7141 cgcggttctt gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct
     7201 ggccggggct ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc
     7261 caagggctgt agtctgggtc cgcgagcaag gttgccctga actgggggtt ggggggagcg
     7321 cagcaaaatg gcggctgttc ccgagtcttg aatggaagac gcttgtgagg cgggctgtga
     7381 ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga acccaaggtc ttgagccctt
     7441 cgctaatgcg ggaaagctct tattcgggtg agatgggctg ggcaccatct ggggaccctg
     7501 acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt
     7561 atggcggtgc cgttgggcag tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg
     7621 tgacgtcacc cgttctgttg gcttataatg cagggtgggg ccacctgccg gtaggtgtgc
     7681 ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct agggtaggct ctcctgaatc
     7741 gacaggcgcc ggacctctgg tgaggggagg gataagtgag gcgtcagttt ctttggtcgg
     7801 ttttatgtac ctatcttctt aagtagctga agctccggtt ttgaactatg cgctcggggt
     7861 tggcgagtgt gttttgtgaa gttttttagg caccttttga aatgtaatca tttgggtcaa
     7921 tatgtaattt tcagtgttag acttgtaaat tgtccgctaa attctggccg tttttggctt
     7981 ttttgttaga caacatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     8041 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     8101 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     8161 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     8221 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     8281 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggagggcctc ctggcgccgc
     8341 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     8401 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggccgccgag cgcgccgggg
     8461 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     8521 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     8581 agcccggtgc ctaggctagt atgtaagcct agtcttagat aataaaatcg ctatccatcg
     8641 aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga
     8701 acgatcctct cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg
     8761 aatccagctt gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg
     8821 cagcgtaccg atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga
     8881 gtcctagctt ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt
     8941 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     9001 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg
     9061 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     9121 cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     9181 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     9241 tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     9301 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga
     9361 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     9421 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     9481 gcaatggcaa caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg
     9541 caacagttga tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     9601 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     9661 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     9721 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     9781 attaagcatt ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg
     9841 ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct tccccaactt
     9901 gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta
     9961 aactcgactc tggctctatc gaatctccgt cgtttcgagc ttac
//
