LOCUS       sequence-295416-        9997 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 9997)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 9997)
  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..9997
                     /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             3848..3913
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    3966..4087
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3977..3996)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4031..4050
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4202..4248
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4278..4414
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4459..4530
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4756..5344)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4836..4855)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5844..7053
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6890..6909
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             7057..7656
                     /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(7057..7076)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7553..7573
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7683..7731
                     /note="synthetic polyadenylation signal"
     CDS             join(7997..8065,8066..8857)
                     /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(8215..8234)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      8870..9006
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    9151..9222
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(9227..9273)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    9291..9339
                     /note="synthetic polyadenylation signal"
     enhancer        9420..9723
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9725..9927
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9878..9898
                     /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 gtcgatggat tataaggacc acgatggaga ctacaaagat catgacattg attacaagga
     3901 cgacgatgac aaataagcta gcttgactga ctgagataca gcgtaccttc agctcacaga
     3961 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     4021 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     4081 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     4141 ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc
     4201 gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc
     4261 cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     4321 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     4381 atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc
     4441 tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga
     4501 agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac
     4561 ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc
     4621 tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac
     4681 gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4741 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4801 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4861 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4921 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4981 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5041 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5101 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5161 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5221 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5281 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5341 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5401 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5461 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5521 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5581 atttaaattt ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat
     5641 ccatcttgca ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct
     5701 tggctattgc ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc
     5761 gggatcaccc gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg
     5821 aatatcgaaa tcggggcgcg cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc
     5881 ccccctcgtc acggcgagcg ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct
     5941 tccgcccgga cgctcaggac agcggcccgc tgctcataag actcggcctt agaaccccag
     6001 tatcagcaga aggacatttt aggacgggac ttgggtgact ctagggcact ggttttcttt
     6061 ccagagagcg gaacaggcga ggaaaagtag tcccttctcg gcgattctgc ggagggatct
     6121 ccgtggggcg gtgaacgccg atgattatat aaggacgcgc cgggtgtggc acagctagtt
     6181 ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg
     6241 gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc cgccgggccg ctcggtggga
     6301 cggaagcgtg tggagagacc gccaagggct gtagtctggg tccgcgagca aggttgccct
     6361 gaactggggg ttggggggag cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag
     6421 acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa
     6481 gaacccaagg tcttgagccc ttcgctaatg cgggaaagct cttattcggg tgagatgggc
     6541 tgggcaccat ctggggaccc tgacgtgaag tttgtcactg actggagaac tcggtttgtc
     6601 gtctgttgcg ggggcggcag ttatggcggt gccgttgggc agtgcacccg tacctttggg
     6661 agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa tgcagggtgg
     6721 ggccacctgc cggtaggtgt gcggtaggct tttctccgtc gcaggacgca gggttcgggc
     6781 ctagggtagg ctctcctgaa tcgacaggcg ccggacctct ggtgagggga gggataagtg
     6841 aggcgtcagt ttctttggtc ggttttatgt acctatcttc ttaagtagct gaagctccgg
     6901 ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg aagtttttta ggcacctttt
     6961 gaaatgtaat catttgggtc aatatgtaat tttcagtgtt agacttgtaa attgtccgct
     7021 aaattctggc cgtttttggc ttttttgtta gacaacatga ccgagtacaa gcccacggtg
     7081 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
     7141 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     7201 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     7261 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     7321 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     7381 atggagggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     7441 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     7501 gaggccgccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     7561 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     7621 cgcacctggt gcatgacccg caagcccggt gcctaggcta gtatgtaagc ctagtcttag
     7681 ataataaaat cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgct
     7741 aggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga tccatatcgt
     7801 tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa tcgtcagata
     7861 ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat attgatagtc
     7921 tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag agacaggatc
     7981 agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     8041 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     8101 agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     8161 tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag ttctgctatg
     8221 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     8281 ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat
     8341 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     8401 acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     8461 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     8521 gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat taactggcga
     8581 actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg ataaagttgc
     8641 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     8701 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     8761 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     8821 cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg cattggcgca
     8881 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
     8941 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
     9001 ccttaagatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg
     9061 cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct
     9121 tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct
     9181 tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca
     9241 agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa
     9301 accaacacac atccatcttc gatggatagc gattttatta tctaactgct gatcgagtgt
     9361 agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     9421 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     9481 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     9541 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     9601 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     9661 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     9721 atgctgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     9781 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     9841 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     9901 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatcag atctttgtcg
     9961 atcctaccat ccactcgaca cacccgccag cggccgc
//
