LOCUS       sequence-295088-        9973 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 surface glycoprotein 
            (Spike protein).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9973)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 9973)
  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-m4134332
FEATURES             Location/Qualifiers
     source          1..9973
                     /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..3889
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    3942..4063
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3953..3972)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4007..4026
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4178..4224
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4254..4390
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4435..4506
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4732..5320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4812..4831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5820..7029
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6866..6885
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             7033..7632
                     /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(7033..7052)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7529..7549
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7659..7707
                     /note="synthetic polyadenylation signal"
     CDS             join(7973..8041,8042..8833)
                     /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(8191..8210)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      8846..8982
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    9127..9198
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(9203..9249)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    9267..9315
                     /note="synthetic polyadenylation signal"
     enhancer        9396..9699
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9701..9903
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9854..9874
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgttc gtctttcttg tgttactccc cctggtttcg tcccagtgcg tgaacttgac
       61 tacgcgaacc caactgccgc cagcatatac aaatagtttc acccgtggag tatactaccc
      121 tgataaagtg tttagatcaa gcgtactaca ctctactcag gacctgtttc tccccttctt
      181 ctccaatgtc acatggtttc atgcgatcca cgttagcggc accaacggga caaagcgctt
      241 tgacaaccct gtgctgccat tcaatgatgg tgtgtatttt gcttctaccg agaagagcaa
      301 cataattcgg ggctggatct tcggaactac cctcgacagt aaaactcagt cattgctgat
      361 tgtgaataac gccacaaatg tcgttatcaa ggtctgtgaa tttcaattct gcaacgatcc
      421 attccttggg gtgtattacc ataaaaataa taagtcctgg atggagagcg aatttagggt
      481 atactcgtct gccaacaatt gtactttcga gtatgtgtct cagcccttct taatggatct
      541 ggaaggtaaa cagggcaact ttaagaacct ccgcgagttt gtattcaaaa atatagacgg
      601 atactttaag atttattcca agcacacacc aatcaatttg gtgcgggatc tcccgcaagg
      661 gttcagcgct ctggaacctc tagtggacct ccccatcggc attaacatca caagattcca
      721 gacgctgtta gccctgcata gatcatacct tactcccggt gacagtagtt ctgggtggac
      781 cgcaggcgcg gctgcatatt acgtgggata tttgcagcct cggacgtttc tgctgaaata
      841 caacgagaac ggcaccatta cagatgccgt ggattgcgcc cttgaccccc taagcgaaac
      901 taagtgtacg cttaaaagct tcaccgtcga gaagggaatt taccagactt ccaactttag
      961 ggtccagccg acagaatcaa tagttaggtt cccaaatatc accaatttgt gcccttttgg
     1021 ggaagtcttc aatgctacac gatttgcttc ggtttatgcc tggaacagga agcgtatctc
     1081 aaattgcgtc gcggactaca gcgtgctcta taatagtgca agtttctcga cctttaaatg
     1141 ttatggcgtg tccccgacca aactaaatga cctctgtttt acaaacgttt acgccgattc
     1201 ttttgtgatt cgcggtgatg aggtacgcca aatcgcaccg ggccagactg gcaagattgc
     1261 cgattacaac tacaagttgc ctgacgactt caccggatgt gttatagcct ggaatagcaa
     1321 taatctggat tctaaggtcg gaggtaacta taactacctg tatagactgt tccgtaagtc
     1381 aaacctgaaa ccctttgagc gggacatctc cacagagatt taccaagcag ggtctacccc
     1441 ctgcaatggg gttgaaggat tcaattgcta ctttccatta cagagctacg ggttccagcc
     1501 tacgaacggg gttggttacc agccttatag ggtagtggtg ctgtcgttcg aactgttgca
     1561 cgctccagct acagtgtgcg ggcctaaaaa atcaactaac ctggttaaaa ataagtgtgt
     1621 taattttaac ttcaatggct taaccggaac tggagtgctc acggagtcaa acaagaaatt
     1681 tctaccgttt cagcaattcg ggagggatat cgcagacact acggacgcgg tcagagatcc
     1741 acagaccctt gagatccttg acataacacc ctgctctttc ggaggtgtct ccgtaattac
     1801 accaggcact aatacctcta atcaggtggc cgtgctttat caggatgtta actgtacaga
     1861 ggtcccggtg gctatccatg ccgaccaatt gacacctacg tggcgcgtct attctaccgg
     1921 ctccaacgtc ttccagaccc gggctggatg cttgataggc gcagaacacg taaacaactc
     1981 ttacgaatgt gatattccca ttggagccgg gatctgcgca agctaccaga cccaaacaaa
     2041 cagtcccaga cgggctcgta gcgtggcctc tcagagcata atagcctata ccatgtccct
     2101 tggcgcggag aatagtgtgg catactcaaa taatagcatc gctatcccca ctaactttac
     2161 tatttccgtg acgactgaga ttctgccagt gtctatgact aagacatccg tggattgcac
     2221 aatgtatatt tgtggggatt ccacagagtg tagcaacttg ttgctgcaat atggatcatt
     2281 ctgtacgcag ctgaatcgcg ccctcaccgg catcgctgtg gaacaggaca agaacactca
     2341 agaggtcttt gctcaggtga agcaaatcta caagacccca cctataaaag acttcggggg
     2401 ttttaacttc tcgcagattc tccccgaccc cagcaagcct tcaaaaagga gctttattga
     2461 ggacctgttg ttcaacaagg ttactttagc cgatgccggt tttatcaaac agtacggcga
     2521 ctgtcttggc gatatagcag cccgggatct gatttgcgca caaaaattca atggccttac
     2581 cgtactgcca ccccttctca ccgacgaaat gatcgcgcag tacacctccg cactgctggc
     2641 tgggaccatc acttcgggat ggacctttgg ggctggtgct gctttgcaaa ttcccttcgc
     2701 catgcagatg gcctatcgat tcaatggcat aggggtaaca cagaacgtat tgtatgaaaa
     2761 tcaaaagcta atcgctaacc agtttaattc agccattggt aagatccagg attcactctc
     2821 cagtacagcc tctgccctgg gaaagctcca ggacgtggtc aaccagaacg cgcaggcatt
     2881 gaatacactc gtcaaacagc tgagtagcaa tttcggcgca atcagtagtg tcttaaacga
     2941 tatactgagt aggcttgata aagtagaggc cgaggtccag attgacagac taatcacggg
     3001 gcgtcttcag agtttacaga cctatgtgac ccaacagctg atccgagcag cggaaattcg
     3061 agcaagcgcg aacctggccg ctactaaaat gagtgagtgc gtgctgggcc aaagtaaacg
     3121 agtcgacttt tgcggcaaag ggtatcatct catgtctttc cctcagtccg cccctcacgg
     3181 tgttgttttc ctccacgtga catacgttcc cgcgcaggaa aagaatttta ctacggcacc
     3241 cgctatttgc catgacggaa aggctcattt tccacgggag ggagtgttcg tttcaaacgg
     3301 gacacactgg tttgtaactc agcggaactt ttatgagcct caaatcatca cgacagacaa
     3361 taccttcgtg agtggcaact gcgatgtagt cataggcatt gtgaataaca ctgtttatga
     3421 tccacttcag ccggaactcg acagcttcaa agaggaattg gataagtact ttaaaaatca
     3481 taccagtcca gacgtagatc tcggcgatat atcgggcatt aacgccagtg tggtcaatat
     3541 acaaaaggag attgacagac tgaatgaagt agctaagaac ctcaacgaat ctctgattga
     3601 cctgcaagag ctagggaagt acgagcagta tatcaaatgg ccatggtaca tatggctggg
     3661 ttttatcgcg ggtcttattg ccatagtcat ggtgacgatt atgttgtgct gtatgacctc
     3721 ctgctgttct tgtttgaaag gctgctgctc ctgtggtagc tgttgcaaat tcgatgaaga
     3781 tgactctgaa cccgtgttga aaggcgttaa gctacactac acagaaaacc tttactttca
     3841 gtcgaaagaa accgcggcag ctaagttcga gaggcagcac atggacagtt aagctagctt
     3901 gactgactga gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt
     3961 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     4021 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     4081 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     4141 ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct
     4201 aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg
     4261 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     4321 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     4381 ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc
     4441 tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc
     4501 tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta
     4561 cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg
     4621 atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc
     4681 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4741 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4801 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4861 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4921 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4981 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5041 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5101 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5161 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5221 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5281 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5341 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5401 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5461 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5521 ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag
     5581 gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa
     5641 cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat
     5701 cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac
     5761 ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg
     5821 gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg cgagcgctgc
     5881 cacgtcagac gaagggcgca ggagcgtcct gatccttccg cccggacgct caggacagcg
     5941 gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga cattttagga
     6001 cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac aggcgaggaa
     6061 aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga acgccgatga
     6121 ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg atttgggtcg
     6181 cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct gctgggctgg
     6241 ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga gagaccgcca
     6301 agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg ggggagcgca
     6361 gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg
     6421 tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt gagcccttcg
     6481 ctaatgcggg aaagctctta ttcgggtgag atgggctggg caccatctgg ggaccctgac
     6541 gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg cggcagttat
     6601 ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg
     6661 acgtcacccg ttctgttggc ttataatgca gggtggggcc acctgccggt aggtgtgcgg
     6721 taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct cctgaatcga
     6781 caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct ttggtcggtt
     6841 ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg ctcggggttg
     6901 gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt tgggtcaata
     6961 tgtaattttc agtgttagac ttgtaaattg tccgctaaat tctggccgtt tttggctttt
     7021 ttgttagaca acatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     7081 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     7141 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     7201 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     7261 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     7321 gagttgagcg gttcccggct ggccgcgcag caacagatgg agggcctcct ggcgccgcac
     7381 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     7441 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg ccgccgagcg cgccggggtg
     7501 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     7561 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     7621 cccggtgcct aggctagtat gtaagcctag tcttagataa taaaatcgct atccatcgaa
     7681 gatggatgtg tgttggtttt ttgtgtgtgt aacgctaggc gcgcctggtg taccgagaac
     7741 gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag ccagtcggaa
     7801 tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct ggtcacggca
     7861 gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt ataatcgagt
     7921 cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc gcatgagtat
     7981 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     8041 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg
     8101 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     8161 ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     8221 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     8281 ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     8341 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga ttggaggacc
     8401 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     8461 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     8521 aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag cttcccggca
     8581 acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     8641 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     8701 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     8761 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     8821 taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct
     8881 gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc
     8941 ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcgttt aaactcgact
     9001 ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc
     9061 tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag cgatcgcggc
     9121 tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg gtcataacag
     9181 cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg
     9241 gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc atcttcgatg
     9301 gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta atcaattacg
     9361 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     9421 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     9481 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     9541 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     9601 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     9661 tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt ttggcagtac
     9721 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     9781 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     9841 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     9901 gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac tcgacacacc
     9961 cgccagcggc cgc
//
