LOCUS       sequence-307368-        9953 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 9953)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 9953)
  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-m4252845
FEATURES             Location/Qualifiers
     source          1..9953
                     /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..3862
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3915..4036
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3926..3945)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3980..3999
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4151..4197
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4227..4363
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4408..4479
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4705..5293)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4785..4804)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5793..7002
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6839..6858
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             7006..7605
                     /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(7006..7025)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7502..7522
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7632..7680
                     /note="synthetic polyadenylation signal"
     CDS             join(7946..8014,8015..8806)
                     /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(8164..8183)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      8819..8955
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    9107..9178
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(9183..9229)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    9247..9295
                     /note="synthetic polyadenylation signal"
     enhancer        9376..9679
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9681..9883
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9834..9854
                     /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 gtcgcaccat caccatcacc attaagctag cttgactgac tgagatacag cgtaccttca
     3901 gctcacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3961 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     4021 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     4081 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct
     4141 atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc
     4201 ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc
     4261 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     4321 cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg
     4381 caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta
     4441 ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc
     4501 tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt
     4561 cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc
     4621 tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4681 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4741 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4801 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4861 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4921 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4981 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5041 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5101 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5161 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5221 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5281 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5341 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5401 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5461 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5521 catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt
     5581 tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc
     5641 cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata
     5701 tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga
     5761 gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc
     5821 cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt
     5881 cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta
     5941 gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg
     6001 gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg
     6061 gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca
     6121 cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat
     6181 cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc
     6241 tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa
     6301 ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt
     6361 gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt
     6421 gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc ttattcgggt
     6481 gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact
     6541 cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt
     6601 acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat
     6661 gcagggtggg gccaccagcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag
     6721 ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag
     6781 ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg
     6841 aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag
     6901 gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gacttgtaaa
     6961 ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatgac cgagtacaag
     7021 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     7081 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     7141 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     7201 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     7261 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     7321 cagcaacaga tggagggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     7381 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     7441 cccggagtgg aggccgccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     7501 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     7561 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctaggctag tatgtaagcc
     7621 tagtcttaga taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg
     7681 tgtaacgcta ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat
     7741 ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat
     7801 cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata
     7861 ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga
     7921 gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc
     7981 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     8041 agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg
     8101 taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt
     8161 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     8221 catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac
     8281 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     8341 ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa
     8401 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     8461 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt
     8521 aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga
     8581 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     8641 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     8701 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     8761 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc
     8821 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca
     8881 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca
     8941 gaaatttatc cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg
     9001 tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg
     9061 tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt
     9121 agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa
     9181 ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc
     9241 gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta
     9301 actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc
     9361 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     9421 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     9481 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     9541 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     9601 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     9661 agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     9721 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     9781 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     9841 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
     9901 gatcagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc cgc
//
