LOCUS       sequence-307360-        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-m4252811
FEATURES             Location/Qualifiers
     source          1..9953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        79..382
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        384..586
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     537..557
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4480..4500
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4501..4518
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4571..4692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4582..4601)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4636..4655
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4807..4853
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4883..5019
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5064..5135
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(5361..5949)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5441..5460)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6449..7658
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     7495..7514
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             7662..8261
                     /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(7662..7681)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8158..8178
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    8288..8336
                     /note="synthetic polyadenylation signal"
     CDS             join(8602..8670,8671..9462)
                     /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(8820..8839)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      9475..9611
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    9763..9834
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(9839..9885)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    9903..9951
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 ctaactgctg atcgagtgta gccagatcta gtaatcaatt acggggtcat tagttcatag
       61 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      121 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      181 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      241 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      301 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      361 attagtcatc gctattacca tgctgatgcg gttttggcag tacatcaatg ggcgtggata
      421 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      481 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      541 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
      601 tcagatcaga tctttgtcga tcctaccatc cactcgacac acccgccagc ggccgccacc
      661 atgtttgttt ttcttgtttt attgccacta gtctctagtc agtgtgttaa tcttacaacc
      721 agaactcaat taccccctgc atacactaat tctttcacac gtggtgttta ttaccctgac
      781 aaagttttca gatcctcagt tttacattca actcaggact tgttcttacc tttcttttcc
      841 aatgttactt ggttccatgc tatacatgtc tctgggacca atggtactaa gaggtttgat
      901 aaccctgtcc taccatttaa tgatggtgtt tattttgctt ccactgagaa gtctaacata
      961 ataagaggct ggatttttgg tactacttta gattcgaaga cccagtccct acttattgtt
     1021 aataacgcta ctaatgttgt tattaaagtc tgtgaatttc aattttgtaa tgatccattt
     1081 ttgggtgttt attaccacaa aaacaacaaa agttggatgg aaagtgagtt cagagtttat
     1141 tctagtgcga ataattgcac ttttgaatat gtctctcagc cttttcttat ggaccttgaa
     1201 ggaaaacagg gtaatttcaa aaatcttagg gaatttgtgt ttaagaatat tgatggttat
     1261 tttaaaatat attctaagca cacgcctatt aatttagtgc gtgatctccc tcagggtttt
     1321 tcggctttag aaccattggt agatttgcca ataggtatta acatcactag gtttcaaact
     1381 ttacttgctt tacatagaag ttatttgact cctggtgatt cttcttcagg ttggacagct
     1441 ggtgctgcag cttattatgt gggttatctt caacctagga cttttctatt aaaatataat
     1501 gaaaatggaa ccattacaga tgctgtagac tgtgcacttg accctctctc agaaacaaag
     1561 tgtacgttga aatccttcac tgtagaaaaa ggaatctatc aaacttctaa ctttagagtc
     1621 caaccaacag aatctattgt tagatttcct aatattacaa acttgtgccc ttttggtgaa
     1681 gtttttaacg ccaccagatt tgcatctgtt tatgcttgga acaggaagag aatcagcaac
     1741 tgtgttgctg attattctgt cctatataat tccgcatcat tttccacttt taagtgttat
     1801 ggagtgtctc ctactaaatt aaatgatctc tgctttacta atgtctatgc agattcattt
     1861 gtaattagag gtgatgaagt cagacaaatc gctccagggc aaactggaaa gattgctgat
     1921 tataattata aattaccaga tgattttaca ggctgcgtta tagcttggaa ttctaacaat
     1981 cttgattcta aggttggtgg taattataat tacctgtata gattgtttag gaagtctaat
     2041 ctcaaacctt ttgagagaga tatttcaact gaaatctatc aggccggtag cacaccttgt
     2101 aatggtgttg aaggttttaa ttgttacttt cctttacaat catatggttt ccaacccact
     2161 aatggtgttg gttaccaacc atacagagta gtagtacttt cttttgaact tctacatgca
     2221 ccagcaactg tttgtggacc taaaaagtct actaatttgg ttaaaaacaa atgtgtcaat
     2281 ttcaacttca atggtttaac aggcacaggt gttcttactg agtctaacaa aaagtttctg
     2341 cctttccaac aatttggcag agacattgct gacactactg atgctgtccg tgatccacag
     2401 acacttgaga ttcttgacat tacaccatgt tcttttggtg gtgtcagtgt tataacacca
     2461 ggaacaaata cttctaacca ggttgctgtt ctttatcagg atgttaactg cacagaagtc
     2521 cctgttgcta ttcatgcaga tcaacttact cctacttggc gtgtttattc tacaggttct
     2581 aatgtttttc aaacacgtgc aggctgttta ataggggctg aacatgtcaa caactcatat
     2641 gagtgtgaca tacccattgg tgcaggtata tgcgctagtt atcagactca gactaattct
     2701 cctcggcggg cacgtagtgt agctagtcaa tccatcattg cctacactat gtcacttggt
     2761 gcagaaaatt cagttgctta ctctaataac tctattgcca tacccacaaa ttttactatt
     2821 agtgttacca cagaaattct accagtgtct atgaccaaga catcagtaga ttgtacaatg
     2881 tacatttgtg gtgattcaac tgaatgcagc aatcttttgt tgcaatatgg cagtttttgt
     2941 acacaattaa accgtgcttt aactggaata gctgttgaac aagacaaaaa cacccaagaa
     3001 gtttttgcac aagtcaaaca aatttacaaa acaccaccaa ttaaagattt tggtggtttt
     3061 aatttttcac aaatattacc agatccatca aaaccaagca agaggtcatt tattgaagat
     3121 ctacttttca acaaagtgac acttgcagat gctggcttca tcaaacaata tggtgattgc
     3181 cttggtgata ttgctgctag agacctcatt tgtgcacaaa agtttaacgg ccttactgtt
     3241 ttgccacctt tgctcacaga tgaaatgatt gctcaataca cttctgcact gttagcgggt
     3301 acaatcactt ctggttggac ctttggtgca ggtgctgcat tacaaatacc atttgctatg
     3361 caaatggctt ataggtttaa tggtattgga gttacacaga atgttctcta tgagaaccaa
     3421 aaattgattg ccaaccaatt taatagtgct attggcaaaa ttcaagactc actttcttcc
     3481 acagcaagtg cacttggaaa acttcaagat gtggtcaacc aaaatgcaca agctttaaac
     3541 acgcttgtta aacaacttag ctccaatttt ggtgcaattt caagtgtttt aaatgatatc
     3601 ctttcacgtc ttgacaaagt tgaggctgaa gtgcaaattg ataggttgat cacaggcaga
     3661 cttcaaagtt tgcagacata tgtgactcaa caattaatta gagctgcaga aatcagagct
     3721 tctgctaatc ttgctgctac taaaatgtca gagtgtgtac ttggacaatc aaaaagagtt
     3781 gatttttgtg gaaagggcta tcatcttatg tccttccctc agtcagcacc tcatggtgta
     3841 gtcttcttgc atgtgactta tgtccctgca caagaaaaga acttcacaac tgctcctgcc
     3901 atttgtcatg atggaaaagc acactttcct cgtgaaggtg tctttgtttc aaatggcaca
     3961 cactggtttg taacacaaag gaatttttat gaaccacaaa tcattactac agacaacaca
     4021 tttgtgtctg gtaactgtga tgttgtaata ggaattgtca acaacacagt ttatgatcct
     4081 ttgcaacctg aattagactc attcaaggag gagttagata aatattttaa gaatcataca
     4141 tcaccagatg ttgatttagg tgacatctct ggcattaatg cttcagttgt aaacattcaa
     4201 aaagaaattg accgcctcaa tgaggttgcc aagaatttaa atgaatctct catcgatctc
     4261 caagaacttg gaaagtatga gcagtatata aaatggccat ggtacatttg gctaggtttt
     4321 atagctggct tgattgccat agtaatggtg acaattatgc tttgctgtat gaccagttgc
     4381 tgtagttgtc tcaagggctg ttgttcttgt ggatcctgct gcaaatttga tgaagacgac
     4441 tctgagccag tgctcaaagg agtcaaatta cattacacag aaaaccttta ctttcagtcg
     4501 caccatcacc atcaccatta agctagcttg actgactgag atacagcgta ccttcagctc
     4561 acagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     4621 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     4681 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     4741 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg
     4801 gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct
     4861 cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     4921 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     4981 cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg
     5041 cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg
     5101 tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc
     5161 gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac
     5221 tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg
     5281 attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5341 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5401 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5461 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5521 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5581 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5641 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5701 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5761 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5821 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5881 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5941 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6001 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6061 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6121 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6181 gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt
     6241 ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg
     6301 cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg
     6361 agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc
     6421 cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg
     6481 ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg
     6541 atccttccgc ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac
     6601 cccagtatca gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt
     6661 tctttccaga gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg
     6721 gatctccgtg gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc
     6781 tagttccgtc gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc
     6841 acttggtgag tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg
     6901 tgggacggaa gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt
     6961 gccctgaact gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat
     7021 ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc
     7081 ggcaagaacc caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga
     7141 tgggctgggc accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt
     7201 ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct
     7261 ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag
     7321 ggtggggcca ccagccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt
     7381 cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat
     7441 aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc
     7501 tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac
     7561 cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt
     7621 ccgctaaatt ctggccgttt ttggcttttt tgttagacaa catgaccgag tacaagccca
     7681 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     7741 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     7801 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     7861 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     7921 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     7981 aacagatgga gggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     8041 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     8101 gagtggaggc cgccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     8161 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     8221 gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta ggctagtatg taagcctagt
     8281 cttagataat aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta
     8341 acgctaggcg cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat
     8401 atcgttgctt ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc
     8461 agatattgta ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga
     8521 tagtctgatc ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca
     8581 ggatcagcag gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt
     8641 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     8701 gctgaagatc agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag
     8761 atccttgaga gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg
     8821 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     8881 cactattctc agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat
     8941 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     9001 aacttacttc tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg
     9061 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     9121 gacgagcgtg acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact
     9181 ggcgaactac ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa
     9241 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     9301 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     9361 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     9421 cagatcgctg agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg
     9481 gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt
     9541 cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa
     9601 tttatcctta agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt
     9661 ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag
     9721 ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct
     9781 ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca
     9841 aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta
     9901 cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat tat
//
