LOCUS       sequence-300710-        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-m4252836
FEATURES             Location/Qualifiers
     source          1..9980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1778..1980)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1807..1827)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1982..2285
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    2366..2414
                     /note="synthetic polyadenylation signal"
     terminator      2432..2478
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    2483..2554
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2706..2842)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(2855..3646,3647..3715))
                     /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     3478..3497
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    3981..4029
                     /note="synthetic polyadenylation signal"
     CDS             complement(4056..4655)
                     /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(4139..4159)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4636..4655
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(4659..5868)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(4803..4822)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      6368..6956
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6857..6876
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7182..7253
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7298..7434)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(7464..7510)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7625..7746
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7662..7681)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7716..7735
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(7799..7843)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(7844..7864)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
ORIGIN
        1 tctctgccaa attgttggaa aggcagaaac tttttgttag actcagtaag aacacctgtg
       61 cctgttaaac cattgaagtt gaaattgaca catttgtttt taaccaaatt agtagacttt
      121 ttaggtccac aaacagttgc tggtgcatgt agaagttcaa aagaaagtac tactactctg
      181 tatggttggt aaccaacacc attagtgggt tggaaaccat atgattgtaa aggaaagtaa
      241 caattaaaac cttcaacacc attacaaggt gtgctaccgg cctgatagat ttcagttgaa
      301 atatctctct caaaaggttt gagattagac ttcctaaaca atctatacag gtaattataa
      361 ttaccaccaa ccttagaatc aagattgtta gaattccaag ctataacgca gcctgtaaaa
      421 tcatctggta atttataatt ataatcagca atctttccag tttgccctgg agcgatttgt
      481 ctgacttcat cacctctaat tacaaatgaa tctgcataga cattagtaaa gcagagatca
      541 tttaatttag taggagacac tccataacac ttaaaagtgg aaaatgatgc ggaattatat
      601 aggacagaat aatcagcaac acagttgctg attctcttcc tgttccaagc ataaacagat
      661 gcaaatctgg tggcgttaaa aacttcacca aaagggcaca agtttgtaat attaggaaat
      721 ctaacaatag attctgttgg ttggactcta aagttagaag tttgatagat tcctttttct
      781 acagtgaagg atttcaacgt acactttgtt tctgagagag ggtcaagtgc acagtctaca
      841 gcatctgtaa tggttccatt ttcattatat tttaatagaa aagtcctagg ttgaagataa
      901 cccacataat aagctgcagc accagctgtc caacctgaag aagaatcacc aggagtcaaa
      961 taacttctat gtaaagcaag taaagtttga aacctagtga tgttaatacc tattggcaaa
     1021 tctaccaatg gttctaaagc cgaaaaaccc tgagggagat cacgcactaa attaataggc
     1081 gtgtgcttag aatatatttt aaaataacca tcaatattct taaacacaaa ttccctaaga
     1141 tttttgaaat taccctgttt tccttcaagg tccataagaa aaggctgaga gacatattca
     1201 aaagtgcaat tattcgcact agaataaact ctgaactcac tttccatcca acttttgttg
     1261 tttttgtggt aataaacacc caaaaatgga tcattacaaa attgaaattc acagacttta
     1321 ataacaacat tagtagcgtt attaacaata agtagggact gggtcttcga atctaaagta
     1381 gtaccaaaaa tccagcctct tattatgtta gacttctcag tggaagcaaa ataaacacca
     1441 tcattaaatg gtaggacagg gttatcaaac ctcttagtac cattggtccc agagacatgt
     1501 atagcatgga accaagtaac attggaaaag aaaggtaaga acaagtcctg agttgaatgt
     1561 aaaactgagg atctgaaaac tttgtcaggg taataaacac cacgtgtgaa agaattagtg
     1621 tatgcagggg gtaattgagt tctggttgta agattaacac actgactaga gactagtggc
     1681 aataaaacaa gaaaaacaaa catggtggcg gccgctggcg ggtgtgtcga gtggatggta
     1741 ggatcgacaa agatctgatc tgacggttca ctaaaccagc tctgcttata tagacctccc
     1801 accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa
     1861 agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg
     1921 gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca
     1981 gcatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg
     2041 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
     2101 tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
     2161 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
     2221 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
     2281 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactagatct
     2341 ggctacactc gatcagcagt tagataataa aatcgctatc catcgaagat ggatgtgtgt
     2401 tggttttttg tgtgtgtaac gcaacgattg atagcataac cccttggggc ctctaaacgg
     2461 gtcttgaggg gttttttgaa ttgagaggta gctgaagctg ctgttatgac cactagaggg
     2521 aagaagatac ctgtggagct aatggtccaa gatgtcggga gccgcgatcg ctgccaaaaa
     2581 ggtaagctga cgatagctga cgagattcga tgcccgacga gcaaatgagc gccacggctg
     2641 ttcgcgtaag ctcgaaacga cggagattcg atagagccag agtcgagttt aaacgattcg
     2701 ggatcttaag gataaatttc tggtaaggag gacacgtatg gaagtgggca agttggggaa
     2761 gccgtatccg ttgctgaatc tggcatatgt gggagtataa gacgcgcagc gtcgcatcag
     2821 gcattttttt ctgcgccaat gcacctagaa tcggttacca atgcttaatc agtgaggcac
     2881 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     2941 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3001 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3061 gaagtggtcc tgcaacttta tccgcctcca tccagtctat caactgttgc cgggaagcta
     3121 gagtaagtag ttcgccagtt aatagtttac gcaaggttgt tgccattgct acaggcatcg
     3181 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3241 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccaatcg
     3301 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3361 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgaatac tcaaccaagt
     3421 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3481 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaagcgt tcttcggggc
     3541 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgcgcac
     3601 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3661 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatgcgaa
     3721 agcctcctgc tgatcctgtc tcttgataga tgtttgcaaa agctaggact cgattatacg
     3781 ttgaccgatc agactatcaa tatctaggtt taaacagatc ggtacgctgc cgtgaccagg
     3841 cttgagtaca atatctgacg attggacttc ctgggtccca agctggattc cgactggctg
     3901 actgccaagc aacgatatgg atcgtcgaga ctcgcactga gaggatcgtt ctcggtacac
     3961 caggcgcgcc tagcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc
     4021 gattttatta tctaagacta ggcttacata ctagcctagg caccgggctt gcgggtcatg
     4081 caccaggtgc gcggtccttc gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc
     4141 tcgtagaagg ggaggttgcg gggcgcggag gtctccagga aggcgggcac cccggcgcgc
     4201 tcggcggcct ccactccggg gagcacgacg gcgctgccca gacccttgcc ctggtggtcg
     4261 ggcgagacgc cgacggtggc caggaaccac gcgggctcct tgggccggtg cggcgccagg
     4321 aggccctcca tctgttgctg cgcggccagc cgggaaccgc tcaactcggc catgcgcggg
     4381 ccgatctcgg cgaacaccgc ccccgcttcg acgctctccg gcgtggtcca gaccgccacc
     4441 gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag
     4501 agttcttgca gctcggtgac ccgctcgatg tggcggtccg gatcgacggt gtggcgcgtg
     4561 gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg ccctggggac gtcgtcgcgg
     4621 gtggcgaggc gcaccgtggg cttgtactcg gtcatgttgt ctaacaaaaa agccaaaaac
     4681 ggccagaatt tagcggacaa tttacaagtc taacactgaa aattacatat tgacccaaat
     4741 gattacattt caaaaggtgc ctaaaaaact tcacaaaaca cactcgccaa ccccgagcgc
     4801 atagttcaaa accggagctt cagctactta agaagatagg tacataaaac cgaccaaaga
     4861 aactgacgcc tcacttatcc ctcccctcac cagaggtccg gcgcctgtcg attcaggaga
     4921 gcctacccta ggcccgaacc ctgcgtcctg cgacggagaa aagcctaccg cacacctacc
     4981 ggctggtggc cccaccctgc attataagcc aacagaacgg gtgacgtcac gacacgacga
     5041 gggcgcgcgc tcccaaaggt acgggtgcac tgcccaacgg caccgccata actgccgccc
     5101 ccgcaacaga cgacaaaccg agttctccag tcagtgacaa acttcacgtc agggtcccca
     5161 gatggtgccc agcccatctc acccgaataa gagctttccc gcattagcga agggctcaag
     5221 accttgggtt cttgccgccc accatgcccc ccaccttgtt tcaacgacct cacagcccgc
     5281 ctcacaagcg tcttccattc aagactcggg aacagccgcc attttgctgc gctcccccca
     5341 acccccagtt cagggcaacc ttgctcgcgg acccagacta cagcccttgg cggtctctcc
     5401 acacgcttcc gtcccaccga gcggcccggc ggccacgaaa gccccggcca gcccagcagc
     5461 ccgctactca ccaagtgacg atcacagcga tccacaaaca agaaccgcga cccaaatccc
     5521 ggctgcgacg gaactagctg tgccacaccc ggcgcgtcct tatataatca tcggcgttca
     5581 ccgccccacg gagatccctc cgcagaatcg ccgagaaggg actacttttc ctcgcctgtt
     5641 ccgctctctg gaaagaaaac cagtgcccta gagtcaccca agtcccgtcc taaaatgtcc
     5701 ttctgctgat actggggttc taaggccgag tcttatgagc agcgggccgc tgtcctgagc
     5761 gtccgggcgg aaggatcagg acgctcctgc gcccttcgtc tgacgtggca gcgctcgccg
     5821 tgacgagggg ggcgcccgcg ggaggcgcca aaacccggcg cggaggccag gcgcgccccg
     5881 atttcgatat tcctcggatc tcggatagat cgggattgag gatgtaggtt gaacttctct
     5941 cgggtgatcc cgatctcgga tattcgggat tggagtaata cctggcgata ggcgctgcca
     6001 agcaatagcc aaggcgcaag gccggccaag agacttgcga tagttcgttc gagaggtagc
     6061 ctgcaagatg gatcggacga aaggtttgaa gattttccga cgagggcctt ggagagttcg
     6121 gaaatttaaa tgcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6181 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6241 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6301 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6361 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6421 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6481 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6541 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6601 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6661 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6721 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6781 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6841 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6901 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6961 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgattt
     7021 aaatgaaact cgtaatcgga gaacgggtgc aatagcaagg accagatcga acttgactga
     7081 cgcaattctg agcgtagtcg aaacccagac aagtcgatct cgccaagtag ctgacttgac
     7141 tggttaatta agtttcgata gcccaaggta accaatgaga ttagggaggg aagaaagttt
     7201 gatgaactac ttctgacccc tagtggtgtc cagaaaagac cattaaagga atgtcttgat
     7261 cgaaatcgag agatcgcctg caggatagct gacgacctta aggataaatt tctggtaagg
     7321 aggacacgta tggaagtggg caagttgggg aagccgtatc cgttgctgaa tctggcatat
     7381 gtgggagtat aagacgcgca gcgtcgcatc aggcattttt ttctgcgcca atgccggtag
     7441 atagcaatcc ggcccgaggg gcacaaaaaa cccctcaaga cccgtttaga ggccccaagg
     7501 ggttatgcta cgataccgat agagatgggc caataccaca tttgtagagg ttttacttgc
     7561 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     7621 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     7681 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     7741 atcttatcat gtctgtgagc tgaaggtacg ctgtatctca gtcagtcaag ctagcttaac
     7801 tgtccatgtg ctgcctctcg aacttagctg ccgcggtttc tttcgactga aagtaaaggt
     7861 tttctgtgta atgtaatttg actcctttga gcactggctc agagtcgtct tcatcaaatt
     7921 tgcagcagga tccacaagaa caacagccct tgagacaact acagcaactg gtcatacagc
     7981 aaagcataat tgtcaccatt actatggcaa tcaagccagc tataaaacct agccaaatgt
     8041 accatggcca ttttatatac tgctcatact ttccaagttc ttggagatcg atgagagatt
     8101 catttaaatt cttggcaacc tcattgaggc ggtcaatttc tttttgaatg tttacaactg
     8161 aagcattaat gccagagatg tcacctaaat caacatctgg tgatgtatga ttcttaaaat
     8221 atttatctaa ctcctccttg aatgagtcta attcaggttg caaaggatca taaactgtgt
     8281 tgttgacaat tcctattaca acatcacagt taccagacac aaatgtgttg tctgtagtaa
     8341 tgatttgtgg ttcataaaaa ttcctttgtg ttacaaacca gtgtgtgcca tttgaaacaa
     8401 agacaccttc acgaggaaag tgtgcttttc catcatgaca aatggcagga gcagttgtga
     8461 agttcttttc ttgtgcaggg acataagtca catgcaagaa gactacacca tgaggtgctg
     8521 actgagggaa ggacataaga tgatagccct ttccacaaaa atcaactctt tttgattgtc
     8581 caagtacaca ctctgacatt ttagtagcag caagattagc agaagctctg atttctgcag
     8641 ctctaattaa ttgttgagtc acatatgtct gcaaactttg aagtctgcct gtgatcaacc
     8701 tatcaatttg cacttcagcc tcaactttgt caagacgtga aaggatatca tttaaaacac
     8761 ttgaaattgc accaaaattg gagctaagtt gtttaacaag cgtgtttaaa gcttgtgcat
     8821 tttggttgac cacatcttga agttttccaa gtgcacttgc tgtggaagaa agtgagtctt
     8881 gaattttgcc aatagcacta ttaaattggt tggcaatcaa tttttggttc tcatagagaa
     8941 cattctgtgt aactccaata ccattaaacc tataagccat ttgcatagca aatggtattt
     9001 gtaatgcagc acctgcacca aaggtccaac cagaagtgat tgtacccgct aacagtgcag
     9061 aagtgtattg agcaatcatt tcatctgtga gcaaaggtgg caaaacagta aggccgttaa
     9121 acttttgtgc acaaatgagg tctctagcag caatatcacc aaggcaatca ccatattgtt
     9181 tgatgaagcc agcatctgca agtgtcactt tgttgaaaag tagatcttca ataaatgacc
     9241 tcttgcttgg ttttgatgga tctggtaata tttgtgaaaa attaaaacca ccaaaatctt
     9301 taattggtgg tgttttgtaa atttgtttga cttgtgcaaa aacttcttgg gtgtttttgt
     9361 cttgttcaac agctattcca gttaaagcac ggtttaattg tgtacaaaaa ctgccatatt
     9421 gcaacaaaag attgctgcat tcagttgaat caccacaaat gtacattgta caatctactg
     9481 atgtcttggt catagacact ggtagaattt ctgtggtaac actaatagta aaatttgtgg
     9541 gtatggcaat agagttatta gagtaagcaa ctgaattttc tgcaccaagt gacatagtgt
     9601 aggcaatgat ggattgacta gctacactac gtgcccgccg aggagaatta gtctgagtct
     9661 gataactagc gcatatacct gcaccaatgg gtatgtcaca ctcatatgag ttgttgacat
     9721 gttcagcccc tattaaacag cctgcacgtg tttgaaaaac attagaacct gtagaataaa
     9781 cacgccaagt aggagtaagt tgatctgcat gaatagcaac agggacttct gtgcagttaa
     9841 catcctgata aagaacagca acctggttag aagtatttgt tcctggtgtt ataacactga
     9901 caccaccaaa agaacatggt gtaatgtcaa gaatctcaag tgtctgtgga tcacggacag
     9961 catcagtagt gtcagcaatg
//
