LOCUS       sequence-301678-        9977 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 9977)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 9977)
  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-m4252789
FEATURES             Location/Qualifiers
     source          1..9977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3730..3750
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3751..3792
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    3845..3966
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3856..3875)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3910..3929
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4081..4127
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4157..4293
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4338..4409
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4635..5223)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4715..4734)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5723..6932
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6769..6788
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             6936..7535
                     /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(6936..6955)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7432..7452
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7562..7610
                     /note="synthetic polyadenylation signal"
     CDS             join(7876..7944,7945..8736)
                     /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(8094..8113)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      8749..8885
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    9037..9108
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(9113..9159)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    9177..9225
                     /note="synthetic polyadenylation signal"
     enhancer        9306..9609
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9611..9813
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9764..9784
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 agtttcaccc gtggagtata ctaccctgat aaagtgttta gatcaagcgt actacactct
       61 actcaggacc tgtttctccc cttcttctcc aatgtcacat ggtttcatgc gatccacgtt
      121 agcggcacca acgggacaaa gcgctttgac aaccctgtgc tgccattcaa tgatggtgtg
      181 tattttgctt ctaccgagaa gagcaacata attcggggct ggatcttcgg aactaccctc
      241 gacagtaaaa ctcagtcatt gctgattgtg aataacgcca caaatgtcgt tatcaaggtc
      301 tgtgaatttc aattctgcaa cgatccattc cttggggtgt attaccataa aaataataag
      361 tcctggatgg agagcgaatt tagggtatac tcgtctgcca acaattgtac tttcgagtat
      421 gtgtctcagc ccttcttaat ggatctggaa ggtaaacagg gcaactttaa gaacctccgc
      481 gagtttgtat tcaaaaatat agacggatac tttaagattt attccaagca cacaccaatc
      541 aatttggtgc gggatctccc gcaagggttc agcgctctgg aacctctagt ggacctcccc
      601 atcggcatta acatcacaag attccagacg ctgttagccc tgcatagatc ataccttact
      661 cccggtgaca gtagttctgg gtggaccgca ggcgcggctg catattacgt gggatatttg
      721 cagcctcgga cgtttctgct gaaatacaac gagaacggca ccattacaga tgccgtggat
      781 tgcgcccttg accccctaag cgaaactaag tgtacgctta aaagcttcac cgtcgagaag
      841 ggaatttacc agacttccaa ctttagggtc cagccgacag aatcaatagt taggttccca
      901 aatatcacca atttgtgccc ttttggggaa gtcttcaatg ctacacgatt tgcttcggtt
      961 tatgcctgga acaggaagcg tatctcaaat tgcgtcgcgg actacagcgt gctctataat
     1021 agtgcaagtt tctcgacctt taaatgttat ggcgtgtccc cgaccaaact aaatgacctc
     1081 tgttttacaa acgtttacgc cgattctttt gtgattcgcg gtgatgaggt acgccaaatc
     1141 gcaccgggcc agactggcaa gattgccgat tacaactaca agttgcctga cgacttcacc
     1201 ggatgtgtta tagcctggaa tagcaataat ctggattcta aggtcggagg taactataac
     1261 tacctgtata gactgttccg taagtcaaac ctgaaaccct ttgagcggga catctccaca
     1321 gagatttacc aagcagggtc taccccctgc aatggggttg aaggattcaa ttgctacttt
     1381 ccattacaga gctacgggtt ccagcctacg aacggggttg gttaccagcc ttatagggta
     1441 gtggtgctgt cgttcgaact gttgcacgct ccagctacag tgtgcgggcc taaaaaatca
     1501 actaacctgg ttaaaaataa gtgtgttaat tttaacttca atggcttaac cggaactgga
     1561 gtgctcacgg agtcaaacaa gaaatttcta ccgtttcagc aattcgggag ggatatcgca
     1621 gacactacgg acgcggtcag agatccacag acccttgaga tccttgacat aacaccctgc
     1681 tctttcggag gtgtctccgt aattacacca ggcactaata cctctaatca ggtggccgtg
     1741 ctttatcagg atgttaactg tacagaggtc ccggtggcta tccatgccga ccaattgaca
     1801 cctacgtggc gcgtctattc taccggctcc aacgtcttcc agacccgggc tggatgcttg
     1861 ataggcgcag aacacgtaaa caactcttac gaatgtgata ttcccattgg agccgggatc
     1921 tgcgcaagct accagaccca aacaaacagt cccagacggg ctcgtagcgt ggcctctcag
     1981 agcataatag cctataccat gtcccttggc gcggagaata gtgtggcata ctcaaataat
     2041 agcatcgcta tccccactaa ctttactatt tccgtgacga ctgagattct gccagtgtct
     2101 atgactaaga catccgtgga ttgcacaatg tatatttgtg gggattccac agagtgtagc
     2161 aacttgttgc tgcaatatgg atcattctgt acgcagctga atcgcgccct caccggcatc
     2221 gctgtggaac aggacaagaa cactcaagag gtctttgctc aggtgaagca aatctacaag
     2281 accccaccta taaaagactt cgggggtttt aacttctcgc agattctccc cgaccccagc
     2341 aagccttcaa aaaggagctt tattgaggac ctgttgttca acaaggttac tttagccgat
     2401 gccggtttta tcaaacagta cggcgactgt cttggcgata tagcagcccg ggatctgatt
     2461 tgcgcacaaa aattcaatgg ccttaccgta ctgccacccc ttctcaccga cgaaatgatc
     2521 gcgcagtaca cctccgcact gctggctggg accatcactt cgggatggac ctttggggct
     2581 ggtgctgctt tgcaaattcc cttcgccatg cagatggcct atcgattcaa tggcataggg
     2641 gtaacacaga acgtattgta tgaaaatcaa aagctaatcg ctaaccagtt taattcagcc
     2701 attggtaaga tccaggattc actctccagt acagcctctg ccctgggaaa gctccaggac
     2761 gtggtcaacc agaacgcgca ggcattgaat acactcgtca aacagctgag tagcaatttc
     2821 ggcgcaatca gtagtgtctt aaacgatata ctgagtaggc ttgataaagt agaggccgag
     2881 gtccagattg acagactaat cacggggcgt cttcagagtt tacagaccta tgtgacccaa
     2941 cagctgatcc gagcagcgga aattcgagca agcgcgaacc tggccgctac taaaatgagt
     3001 gagtgcgtgc tgggccaaag taaacgagtc gacttttgcg gcaaagggta tcatctcatg
     3061 tctttccctc agtccgcccc tcacggtgtt gttttcctcc acgtgacata cgttcccgcg
     3121 caggaaaaga attttactac ggcacccgct atttgccatg acggaaaggc tcattttcca
     3181 cgggagggag tgttcgtttc aaacgggaca cactggtttg taactcagcg gaacttttat
     3241 gagcctcaaa tcatcacgac agacaatacc ttcgtgagtg gcaactgcga tgtagtcata
     3301 ggcattgtga ataacactgt ttatgatcca cttcagccgg aactcgacag cttcaaagag
     3361 gaattggata agtactttaa aaatcatacc agtccagacg tagatctcgg cgatatatcg
     3421 ggcattaacg ccagtgtggt caatatacaa aaggagattg acagactgaa tgaagtagct
     3481 aagaacctca acgaatctct gattgacctg caagagctag ggaagtacga gcagtatatc
     3541 aaatggccat ggtacatatg gctgggtttt atcgcgggtc ttattgccat agtcatggtg
     3601 acgattatgt tgtgctgtat gacctcctgc tgttcttgtt tgaaaggctg ctgctcctgt
     3661 ggtagctgtt gcaaattcga tgaagatgac tctgaacccg tgttgaaagg cgttaagcta
     3721 cactacacag aaaaccttta ctttcagtcg gggaaaccta tacccaatcc acttctcggc
     3781 ttagattcta cctaagctag cttgactgac tgagatacag cgtaccttca gctcacagac
     3841 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     3901 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     3961 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     4021 gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg
     4081 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc
     4141 ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
     4201 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
     4261 tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct
     4321 ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa
     4381 gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact
     4441 taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct
     4501 cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg
     4561 agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4621 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4681 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4741 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4801 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4861 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4921 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4981 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5041 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5101 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5161 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5221 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5281 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5341 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5401 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca
     5461 tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc
     5521 catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt
     5581 ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg
     5641 ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga
     5701 atatcgaaat cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc
     5761 cccctcgtca cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt
     5821 ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt
     5881 atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc
     5941 cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc
     6001 cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc
     6061 cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg
     6121 tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac
     6181 ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg
     6241 aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga
     6301 cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag
     6361 aacccaaggt cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct
     6421 gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg
     6481 tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga
     6541 gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg
     6601 gccaccagcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc
     6661 tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga
     6721 ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt
     6781 tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg
     6841 aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta
     6901 aattctggcc gtttttggct tttttgttag acaacatgac cgagtacaag cccacggtgc
     6961 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     7021 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     7081 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     7141 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     7201 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     7261 tggagggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     7321 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     7381 aggccgccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     7441 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     7501 gcacctggtg catgacccgc aagcccggtg cctaggctag tatgtaagcc tagtcttaga
     7561 taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta
     7621 ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt
     7681 gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat
     7741 tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct
     7801 gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca
     7861 gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     7921 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     7981 gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     8041 gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt
     8101 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     8161 tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg
     8221 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     8281 cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     8341 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     8401 cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa
     8461 ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca
     8521 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     8581 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     8641 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     8701 gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag
     8761 aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa
     8821 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc
     8881 cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag
     8941 cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg
     9001 tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag
     9061 gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac
     9121 ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca
     9181 caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc
     9241 gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc atatatggag
     9301 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     9361 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     9421 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     9481 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     9541 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     9601 attaccatgc tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     9661 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     9721 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     9781 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct
     9841 ttgtcgatcc taccatccac tcgacacacc cgccagcggc cgccaccatg ttcgtctttc
     9901 ttgtgttact ccccctggtt tcgtcccagt gcgtgaactt gactacgcga acccaactgc
     9961 cgccagcata tacaaat
//
