LOCUS       sequence-304470-        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-m4252791
FEATURES             Location/Qualifiers
     source          1..9980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    119..190
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(195..241)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    259..307
                     /note="synthetic polyadenylation signal"
     enhancer        388..691
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        693..895
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     846..866
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4789..4809
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4810..4854
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    4907..5028
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4918..4937)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4972..4991
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      5143..5189
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5219..5355
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5400..5471
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(5697..6285)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5777..5796)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6785..7994
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     7831..7850
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             7998..8597
                     /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(7998..8017)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8494..8514
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    8624..8672
                     /note="synthetic polyadenylation signal"
     CDS             join(8938..9006,9007..9798)
                     /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(9156..9175)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      9811..9947
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg
       61 catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca
      121 tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca
      181 gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct
      241 atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga
      301 ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt
      361 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      421 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      481 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      541 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      601 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      661 catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt acatcaatgg
      721 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      781 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      841 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
      901 agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca cccgccagcg
      961 gccgccacca tgttcgtctt tcttgtgtta ctccccctgg tttcgtccca gtgcgtgaac
     1021 ttgactacgc gaacccaact gccgccagca tatacaaata gtttcacccg tggagtatac
     1081 taccctgata aagtgtttag atcaagcgta ctacactcta ctcaggacct gtttctcccc
     1141 ttcttctcca atgtcacatg gtttcatgcg atccacgtta gcggcaccaa cgggacaaag
     1201 cgctttgaca accctgtgct gccattcaat gatggtgtgt attttgcttc taccgagaag
     1261 agcaacataa ttcggggctg gatcttcgga actaccctcg acagtaaaac tcagtcattg
     1321 ctgattgtga ataacgccac aaatgtcgtt atcaaggtct gtgaatttca attctgcaac
     1381 gatccattcc ttggggtgta ttaccataaa aataataagt cctggatgga gagcgaattt
     1441 agggtatact cgtctgccaa caattgtact ttcgagtatg tgtctcagcc cttcttaatg
     1501 gatctggaag gtaaacaggg caactttaag aacctccgcg agtttgtatt caaaaatata
     1561 gacggatact ttaagattta ttccaagcac acaccaatca atttggtgcg ggatctcccg
     1621 caagggttca gcgctctgga acctctagtg gacctcccca tcggcattaa catcacaaga
     1681 ttccagacgc tgttagccct gcatagatca taccttactc ccggtgacag tagttctggg
     1741 tggaccgcag gcgcggctgc atattacgtg ggatatttgc agcctcggac gtttctgctg
     1801 aaatacaacg agaacggcac cattacagat gccgtggatt gcgcccttga ccccctaagc
     1861 gaaactaagt gtacgcttaa aagcttcacc gtcgagaagg gaatttacca gacttccaac
     1921 tttagggtcc agccgacaga atcaatagtt aggttcccaa atatcaccaa tttgtgccct
     1981 tttggggaag tcttcaatgc tacacgattt gcttcggttt atgcctggaa caggaagcgt
     2041 atctcaaatt gcgtcgcgga ctacagcgtg ctctataata gtgcaagttt ctcgaccttt
     2101 aaatgttatg gcgtgtcccc gaccaaacta aatgacctct gttttacaaa cgtttacgcc
     2161 gattcttttg tgattcgcgg tgatgaggta cgccaaatcg caccgggcca gactggcaag
     2221 attgccgatt acaactacaa gttgcctgac gacttcaccg gatgtgttat agcctggaat
     2281 agcaataatc tggattctaa ggtcggaggt aactataact acctgtatag actgttccgt
     2341 aagtcaaacc tgaaaccctt tgagcgggac atctccacag agatttacca agcagggtct
     2401 accccctgca atggggttga aggattcaat tgctactttc cattacagag ctacgggttc
     2461 cagcctacga acggggttgg ttaccagcct tatagggtag tggtgctgtc gttcgaactg
     2521 ttgcacgctc cagctacagt gtgcgggcct aaaaaatcaa ctaacctggt taaaaataag
     2581 tgtgttaatt ttaacttcaa tggcttaacc ggaactggag tgctcacgga gtcaaacaag
     2641 aaatttctac cgtttcagca attcgggagg gatatcgcag acactacgga cgcggtcaga
     2701 gatccacaga cccttgagat ccttgacata acaccctgct ctttcggagg tgtctccgta
     2761 attacaccag gcactaatac ctctaatcag gtggccgtgc tttatcagga tgttaactgt
     2821 acagaggtcc cggtggctat ccatgccgac caattgacac ctacgtggcg cgtctattct
     2881 accggctcca acgtcttcca gacccgggct ggatgcttga taggcgcaga acacgtaaac
     2941 aactcttacg aatgtgatat tcccattgga gccgggatct gcgcaagcta ccagacccaa
     3001 acaaacagtc ccagacgggc tcgtagcgtg gcctctcaga gcataatagc ctataccatg
     3061 tcccttggcg cggagaatag tgtggcatac tcaaataata gcatcgctat ccccactaac
     3121 tttactattt ccgtgacgac tgagattctg ccagtgtcta tgactaagac atccgtggat
     3181 tgcacaatgt atatttgtgg ggattccaca gagtgtagca acttgttgct gcaatatgga
     3241 tcattctgta cgcagctgaa tcgcgccctc accggcatcg ctgtggaaca ggacaagaac
     3301 actcaagagg tctttgctca ggtgaagcaa atctacaaga ccccacctat aaaagacttc
     3361 gggggtttta acttctcgca gattctcccc gaccccagca agccttcaaa aaggagcttt
     3421 attgaggacc tgttgttcaa caaggttact ttagccgatg ccggttttat caaacagtac
     3481 ggcgactgtc ttggcgatat agcagcccgg gatctgattt gcgcacaaaa attcaatggc
     3541 cttaccgtac tgccacccct tctcaccgac gaaatgatcg cgcagtacac ctccgcactg
     3601 ctggctggga ccatcacttc gggatggacc tttggggctg gtgctgcttt gcaaattccc
     3661 ttcgccatgc agatggccta tcgattcaat ggcatagggg taacacagaa cgtattgtat
     3721 gaaaatcaaa agctaatcgc taaccagttt aattcagcca ttggtaagat ccaggattca
     3781 ctctccagta cagcctctgc cctgggaaag ctccaggacg tggtcaacca gaacgcgcag
     3841 gcattgaata cactcgtcaa acagctgagt agcaatttcg gcgcaatcag tagtgtctta
     3901 aacgatatac tgagtaggct tgataaagta gaggccgagg tccagattga cagactaatc
     3961 acggggcgtc ttcagagttt acagacctat gtgacccaac agctgatccg agcagcggaa
     4021 attcgagcaa gcgcgaacct ggccgctact aaaatgagtg agtgcgtgct gggccaaagt
     4081 aaacgagtcg acttttgcgg caaagggtat catctcatgt ctttccctca gtccgcccct
     4141 cacggtgttg ttttcctcca cgtgacatac gttcccgcgc aggaaaagaa ttttactacg
     4201 gcacccgcta tttgccatga cggaaaggct cattttccac gggagggagt gttcgtttca
     4261 aacgggacac actggtttgt aactcagcgg aacttttatg agcctcaaat catcacgaca
     4321 gacaatacct tcgtgagtgg caactgcgat gtagtcatag gcattgtgaa taacactgtt
     4381 tatgatccac ttcagccgga actcgacagc ttcaaagagg aattggataa gtactttaaa
     4441 aatcatacca gtccagacgt agatctcggc gatatatcgg gcattaacgc cagtgtggtc
     4501 aatatacaaa aggagattga cagactgaat gaagtagcta agaacctcaa cgaatctctg
     4561 attgacctgc aagagctagg gaagtacgag cagtatatca aatggccatg gtacatatgg
     4621 ctgggtttta tcgcgggtct tattgccata gtcatggtga cgattatgtt gtgctgtatg
     4681 acctcctgct gttcttgttt gaaaggctgc tgctcctgtg gtagctgttg caaattcgat
     4741 gaagatgact ctgaacccgt gttgaaaggc gttaagctac actacacaga aaacctttac
     4801 tttcagtcga aagaaaccgc ggcagctaag ttcgagaggc agcacatgga cagttaagct
     4861 agcttgactg actgagatac agcgtacctt cagctcacag acatgataag atacattgat
     4921 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     4981 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     5041 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     5101 aacctctaca aatgtggtat tggcccatct ctatcggtat cgtagcataa ccccttgggg
     5161 cctctaaacg ggtcttgagg ggttttttgt gcccctcggg ccggattgct atctaccggc
     5221 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca
     5281 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca
     5341 gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt cgatcaagac
     5401 attcctttaa tggtcttttc tggacaccac taggggtcag aagtagttca tcaaactttc
     5461 ttccctccct aatctcattg gttaccttgg gctatcgaaa cttaattaac cagtcaagtc
     5521 agctacttgg cgagatcgac ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa
     5581 gttcgatctg gtccttgcta ttgcacccgt tctccgatta cgagtttcat ttaaatcatg
     5641 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5701 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5761 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5821 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5881 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5941 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6001 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6061 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6121 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6181 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6241 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6301 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6361 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6421 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6481 cctatctcag cgatctgtct atttcgttca tccatagttg catttaaatt tccgaactct
     6541 ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc aggctacctc
     6601 tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc ttggctattg cttggcagcg
     6661 cctatcgcca ggtattactc caatcccgaa tatccgagat cgggatcacc cgagagaagt
     6721 tcaacctaca tcctcaatcc cgatctatcc gagatccgag gaatatcgaa atcggggcgc
     6781 gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcgt cacggcgagc
     6841 gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc ttccgcccgg acgctcagga
     6901 cagcggcccg ctgctcataa gactcggcct tagaacccca gtatcagcag aaggacattt
     6961 taggacggga cttgggtgac tctagggcac tggttttctt tccagagagc ggaacaggcg
     7021 aggaaaagta gtcccttctc ggcgattctg cggagggatc tccgtggggc ggtgaacgcc
     7081 gatgattata taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg
     7141 ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc gggctgctgg
     7201 gctggccggg gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt gtggagagac
     7261 cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg gttgggggga
     7321 gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg
     7381 tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca agaacccaag gtcttgagcc
     7441 cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctgggcacca tctggggacc
     7501 ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt cgtctgttgc gggggcggca
     7561 gttatggcgg tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg
     7621 tcgtgacgtc acccgttctg ttggcttata atgcagggtg gggccaccag ccggtaggtg
     7681 tgcggtaggc ttttctccgt cgcaggacgc agggttcggg cctagggtag gctctcctga
     7741 atcgacaggc gccggacctc tggtgagggg agggataagt gaggcgtcag tttctttggt
     7801 cggttttatg tacctatctt cttaagtagc tgaagctccg gttttgaact atgcgctcgg
     7861 ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt
     7921 caatatgtaa ttttcagtgt tagacttgta aattgtccgc taaattctgg ccgtttttgg
     7981 cttttttgtt agacaacatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     8041 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     8101 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     8161 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     8221 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     8281 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggagggc ctcctggcgc
     8341 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
     8401 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggccgcc gagcgcgccg
     8461 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     8521 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     8581 gcaagcccgg tgcctaggct agtatgtaag cctagtctta gataataaaa tcgctatcca
     8641 tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc taggcgcgcc tggtgtaccg
     8701 agaacgatcc tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt
     8761 cggaatccag cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca
     8821 cggcagcgta ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat
     8881 cgagtcctag cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg
     8941 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     9001 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga
     9061 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     9121 gaacgctttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     9181 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     9241 gagtattcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     9301 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgattgga
     9361 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     9421 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     9481 gtagcaatgg caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc
     9541 cggcaacagt tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     9601 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     9661 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     9721 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     9781 ctgattaagc attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg
     9841 acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa
     9901 cttgcccact tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt
     9961 ttaaactcga ctctggctct
//
