LOCUS       sequence-301657-        8073 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 8073)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8073)
  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-m4133745
FEATURES             Location/Qualifiers
     source          1..8073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      48..94
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    99..170
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(322..458)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(471..1262,1263..1331))
                     /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     1094..1113
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2077..2665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2566..2585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2891..2962
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3007..3143)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(3173..3219)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3334..3455
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3371..3390)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3425..3444
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3508..3552)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(3553..3573)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(7467..7669)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(7496..7516)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        7671..7974
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    join(8055..8073,1..30)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 gaagatggat gtgtgttggt tttttgtgtg tgtaacgcaa cgattgatag cataacccct
       61 tggggcctct aaacgggtct tgaggggttt tttgaattga gaggtagctg aagctgctgt
      121 tatgaccact agagggaaga agatacctgt ggagctaatg gtccaagatg tcgggagccg
      181 cgatcgctgc caaaaaggta agctgacgat agctgacgag attcgatgcc cgacgagcaa
      241 atgagcgcca cggctgttcg cgtaagctcg aaacgacgga gattcgatag agccagagtc
      301 gagtttaaac gattcgggat cttaaggata aatttctggt aaggaggaca cgtatggaag
      361 tgggcaagtt ggggaagccg tatccgttgc tgaatctggc atatgtggga gtataagacg
      421 cgcagcgtcg catcaggcat ttttttctgc gccaatgcac ctagaatcgg ttaccaatgc
      481 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      541 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      601 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      661 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctatcaac
      721 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttacgcaa ggttgttgcc
      781 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      841 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      901 ttcggtcctc caatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      961 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     1021 gaatactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     1081 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     1141 aagcgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     1201 taacccactc gcgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     1261 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1321 tgaatactca tgcgaaagcc tcctgctgat cctgtctctt gatagatgtt tgcaaaagct
     1381 aggactcgat tatacgttga ccgatcagac tatcaatatc taggtttaaa cagatcggta
     1441 cgctgccgtg accaggcttg agtacaatat ctgacgattg gacttcctgg gtcccaagct
     1501 ggattccgac tggctgactg ccaagcaacg atatggatcg tcgagactcg cactgagagg
     1561 atcgttctcg gtacaccagg cgcgccccga tttcgatatt cctcggatct cggatagatc
     1621 gggattgagg atgtaggttg aacttctctc gggtgatccc gatctcggat attcgggatt
     1681 ggagtaatac ctggcgatag gcgctgccaa gcaatagcca aggcgcaagg ccggccaaga
     1741 gacttgcgat agttcgttcg agaggtagcc tgcaagatgg atcggacgaa aggtttgaag
     1801 attttccgac gagggccttg gagagttcgg aaatttaaat gcaactatgg atgaacgaaa
     1861 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     1921 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     1981 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2041 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2101 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2161 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2221 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2281 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2341 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2401 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2461 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2521 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2581 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     2641 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     2701 cttttgctgg ccttttgctc acatgattta aatgaaactc gtaatcggag aacgggtgca
     2761 atagcaagga ccagatcgaa cttgactgac gcaattctga gcgtagtcga aacccagaca
     2821 agtcgatctc gccaagtagc tgacttgact ggttaattaa gtttcgatag cccaaggtaa
     2881 ccaatgagat tagggaggga agaaagtttg atgaactact tctgacccct agtggtgtcc
     2941 agaaaagacc attaaaggaa tgtcttgatc gaaatcgaga gatcgcctgc aggatagctg
     3001 acgaccttaa ggataaattt ctggtaagga ggacacgtat ggaagtgggc aagttgggga
     3061 agccgtatcc gttgctgaat ctggcatatg tgggagtata agacgcgcag cgtcgcatca
     3121 ggcatttttt tctgcgccaa tgccggtaga tagcaatccg gcccgagggg cacaaaaaac
     3181 ccctcaagac ccgtttagag gccccaaggg gttatgctac gataccgata gagatgggcc
     3241 aataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     3301 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     3361 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3421 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtgagct gaaggtacgc
     3481 tgtatctcag tcagtcaagc tagcttaact gtccatgtgc tgcctctcga acttagctgc
     3541 cgcggtttct ttcgactgaa agtaaaggtt ttctgtgtag tgtagcttaa cgcctttcaa
     3601 cacgggttca gagtcatctt catcgaattt gcaacagcta ccacaggagc agcagccttt
     3661 caaacaagaa cagcaggagg tcatacagca caacataatc gtcaccatga ctatggcaat
     3721 aagacccgcg ataaaaccca gccatatgta ccatggccat ttgatatact gctcgtactt
     3781 ccctagctct tgcaggtcaa tcagagattc gttgaggttc ttagctactt cattcagtct
     3841 gtcaatctcc ttttgtatat tgaccacact ggcgttaatg cccgatatat cgccgagatc
     3901 tacgtctgga ctggtatgat ttttaaagta cttatccaat tcctctttga agctgtcgag
     3961 ttccggctga agtggatcat aaacagtgtt attcacaatg cctatgacta catcgcagtt
     4021 gccactcacg aaggtattgt ctgtcgtgat gatttgaggc tcataaaagt tccgctgagt
     4081 tacaaaccag tgtgtcccgt ttgaaacgaa cactccctcc cgtggaaaat gagcctttcc
     4141 gtcatggcaa atagcgggtg ccgtagtaaa attcttttcc tgcgcgggaa cgtatgtcac
     4201 gtggaggaaa acaacaccgt gaggggcgga ctgagggaaa gacatgagat gatacccttt
     4261 gccgcaaaag tcgactcgtt tactttggcc cagcacgcac tcactcattt tagtagcggc
     4321 caggttcgcg cttgctcgaa tttccgctgc tcggatcagc tgttgggtca cataggtctg
     4381 taaactctga agacgccccg tgattagtct gtcaatctgg acctcggcct ctactttatc
     4441 aagcctactc agtatatcgt ttaagacact actgattgcg ccgaaattgc tactcagctg
     4501 tttgacgagt gtattcaatg cctgcgcgtt ctggttgacc acgtcctgga gctttcccag
     4561 ggcagaggct gtactggaga gtgaatcctg gatcttacca atggctgaat taaactggtt
     4621 agcgattagc ttttgatttt catacaatac gttctgtgtt acccctatgc cattgaatcg
     4681 ataggccatc tgcatggcga agggaatttg caaagcagca ccagccccaa aggtccatcc
     4741 cgaagtgatg gtcccagcca gcagtgcgga ggtgtactgc gcgatcattt cgtcggtgag
     4801 aaggggtggc agtacggtaa ggccattgaa tttttgtgcg caaatcagat cccgggctgc
     4861 tatatcgcca agacagtcgc cgtactgttt gataaaaccg gcatcggcta aagtaacctt
     4921 gttgaacaac aggtcctcaa taaagctcct ttttgaaggc ttgctggggt cggggagaat
     4981 ctgcgagaag ttaaaacccc cgaagtcttt tataggtggg gtcttgtaga tttgcttcac
     5041 ctgagcaaag acctcttgag tgttcttgtc ctgttccaca gcgatgccgg tgagggcgcg
     5101 attcagctgc gtacagaatg atccatattg cagcaacaag ttgctacact ctgtggaatc
     5161 cccacaaata tacattgtgc aatccacgga tgtcttagtc atagacactg gcagaatctc
     5221 agtcgtcacg gaaatagtaa agttagtggg gatagcgatg ctattatttg agtatgccac
     5281 actattctcc gcgccaaggg acatggtata ggctattatg ctctgagagg ccacgctacg
     5341 agcccgtctg ggactgtttg tttgggtctg gtagcttgcg cagatcccgg ctccaatggg
     5401 aatatcacat tcgtaagagt tgtttacgtg ttctgcgcct atcaagcatc cagcccgggt
     5461 ctggaagacg ttggagccgg tagaatagac gcgccacgta ggtgtcaatt ggtcggcatg
     5521 gatagccacc gggacctctg tacagttaac atcctgataa agcacggcca cctgattaga
     5581 ggtattagtg cctggtgtaa ttacggagac acctccgaaa gagcagggtg ttatgtcaag
     5641 gatctcaagg gtctgtggat ctctgaccgc gtccgtagtg tctgcgatat ccctcccgaa
     5701 ttgctgaaac ggtagaaatt tcttgtttga ctccgtgagc actccagttc cggttaagcc
     5761 attgaagtta aaattaacac acttattttt aaccaggtta gttgattttt taggcccgca
     5821 cactgtagct ggagcgtgca acagttcgaa cgacagcacc actaccctat aaggctggta
     5881 accaaccccg ttcgtaggct ggaacccgta gctctgtaat ggaaagtagc aattgaatcc
     5941 ttcaacccca ttgcaggggg tagaccctgc ttggtaaatc tctgtggaga tgtcccgctc
     6001 aaagggtttc aggtttgact tacggaacag tctatacagg tagttatagt tacctccgac
     6061 cttagaatcc agattattgc tattccaggc tataacacat ccggtgaagt cgtcaggcaa
     6121 cttgtagttg taatcggcaa tcttgccagt ctggcccggt gcgatttggc gtacctcatc
     6181 accgcgaatc acaaaagaat cggcgtaaac gtttgtaaaa cagaggtcat ttagtttggt
     6241 cggggacacg ccataacatt taaaggtcga gaaacttgca ctattataga gcacgctgta
     6301 gtccgcgacg caatttgaga tacgcttcct gttccaggca taaaccgaag caaatcgtgt
     6361 agcattgaag acttccccaa aagggcacaa attggtgata tttgggaacc taactattga
     6421 ttctgtcggc tggaccctaa agttggaagt ctggtaaatt cccttctcga cggtgaagct
     6481 tttaagcgta cacttagttt cgcttagggg gtcaagggcg caatccacgg catctgtaat
     6541 ggtgccgttc tcgttgtatt tcagcagaaa cgtccgaggc tgcaaatatc ccacgtaata
     6601 tgcagccgcg cctgcggtcc acccagaact actgtcaccg ggagtaaggt atgatctatg
     6661 cagggctaac agcgtctgga atcttgtgat gttaatgccg atggggaggt ccactagagg
     6721 ttccagagcg ctgaaccctt gcgggagatc ccgcaccaaa ttgattggtg tgtgcttgga
     6781 ataaatctta aagtatccgt ctatattttt gaatacaaac tcgcggaggt tcttaaagtt
     6841 gccctgttta ccttccagat ccattaagaa gggctgagac acatactcga aagtacaatt
     6901 gttggcagac gagtataccc taaattcgct ctccatccag gacttattat ttttatggta
     6961 atacacccca aggaatggat cgttgcagaa ttgaaattca cagaccttga taacgacatt
     7021 tgtggcgtta ttcacaatca gcaatgactg agttttactg tcgagggtag ttccgaagat
     7081 ccagccccga attatgttgc tcttctcggt agaagcaaaa tacacaccat cattgaatgg
     7141 cagcacaggg ttgtcaaagc gctttgtccc gttggtgccg ctaacgtgga tcgcatgaaa
     7201 ccatgtgaca ttggagaaga aggggagaaa caggtcctga gtagagtgta gtacgcttga
     7261 tctaaacact ttatcagggt agtatactcc acgggtgaaa ctatttgtat atgctggcgg
     7321 cagttgggtt cgcgtagtca agttcacgca ctgggacgaa accaggggga gtaacacaag
     7381 aaagacgaac atggtggcgg ccgctggcgg gtgtgtcgag tggatggtag gatcgacaaa
     7441 gatctgatct gacggttcac taaaccagct ctgcttatat agacctccca ccgtacacgc
     7501 ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga
     7561 ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt
     7621 gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcag catggtaata
     7681 gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg
     7741 gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata
     7801 tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt
     7861 caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg
     7921 cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac
     7981 tccatatatg ggctatgaac taatgacccc gtaattgatt actagatctg gctacactcg
     8041 atcagcagtt agataataaa atcgctatcc atc
//
