LOCUS       sequence-300716-        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-m4133806
FEATURES             Location/Qualifiers
     source          1..8073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1619..1821)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1648..1668)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1823..2126
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    2207..2255
                     /note="synthetic polyadenylation signal"
     terminator      2273..2319
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    2324..2395
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2547..2683)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(2696..3487,3488..3556))
                     /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     3319..3338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4302..4890
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4791..4810
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5116..5187
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5232..5368)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5398..5444)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5559..5680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5596..5615)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5650..5669
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5733..5777)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(5778..5798)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
ORIGIN
        1 aaagaaagta ctactactct gtatggttgg taaccaacac cattagtggg ttggaaacca
       61 tatgattgta aaggaaagta acaattaaaa ccttcaacac cattacaagg tgtgctaccg
      121 gcctgataga tttcagttga aatatctctc tcaaaaggtt tgagattaga cttcctaaac
      181 aatctataca ggtaattata attaccacca accttagaat caagattgtt agaattccaa
      241 gctataacgc agcctgtaaa atcatctggt aatttataat tataatcagc aatctttcca
      301 gtttgccctg gagcgatttg tctgacttca tcacctctaa ttacaaatga atctgcatag
      361 acattagtaa agcagagatc atttaattta gtaggagaca ctccataaca cttaaaagtg
      421 gaaaatgatg cggaattata taggacagaa taatcagcaa cacagttgct gattctcttc
      481 ctgttccaag cataaacaga tgcaaatctg gtggcgttaa aaacttcacc aaaagggcac
      541 aagtttgtaa tattaggaaa tctaacaata gattctgttg gttggactct aaagttagaa
      601 gtttgataga ttcctttttc tacagtgaag gatttcaacg tacactttgt ttctgagaga
      661 gggtcaagtg cacagtctac agcatctgta atggttccat tttcattata ttttaataga
      721 aaagtcctag gttgaagata acccacataa taagctgcag caccagctgt ccaacctgaa
      781 gaagaatcac caggagtcaa ataacttcta tgtaaagcaa gtaaagtttg aaacctagtg
      841 atgttaatac ctattggcaa atctaccaat ggttctaaag ccgaaaaacc ctgagggaga
      901 tcacgcacta aattaatagg cgtgtgctta gaatatattt taaaataacc atcaatattc
      961 ttaaacacaa attccctaag atttttgaaa ttaccctgtt ttccttcaag gtccataaga
     1021 aaaggctgag agacatattc aaaagtgcaa ttattcgcac tagaataaac tctgaactca
     1081 ctttccatcc aacttttgtt gtttttgtgg taataaacac ccaaaaatgg atcattacaa
     1141 aattgaaatt cacagacttt aataacaaca ttagtagcgt tattaacaat aagtagggac
     1201 tgggtcttcg aatctaaagt agtaccaaaa atccagcctc ttattatgtt agacttctca
     1261 gtggaagcaa aataaacacc atcattaaat ggtaggacag ggttatcaaa cctcttagta
     1321 ccattggtcc cagagacatg tatagcatgg aaccaagtaa cattggaaaa gaaaggtaag
     1381 aacaagtcct gagttgaatg taaaactgag gatctgaaaa ctttgtcagg gtaataaaca
     1441 ccacgtgtga aagaattagt gtatgcaggg ggtaattgag ttctggttgt aagattaaca
     1501 cactgactag agactagtgg caataaaaca agaaaaacaa acatggtggc ggccgctggc
     1561 gggtgtgtcg agtggatggt aggatcgaca aagatctgat ctgacggttc actaaaccag
     1621 ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg
     1681 agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
     1741 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga
     1801 tgtactgcca aaaccgcatc agcatggtaa tagcgatgac taatacgtag atgtactgcc
     1861 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     1921 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     1981 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     2041 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     2101 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     2161 ccgtaattga ttactagatc tggctacact cgatcagcag ttagataata aaatcgctat
     2221 ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa cgcaacgatt gatagcataa
     2281 ccccttgggg cctctaaacg ggtcttgagg ggttttttga attgagaggt agctgaagct
     2341 gctgttatga ccactagagg gaagaagata cctgtggagc taatggtcca agatgtcggg
     2401 agccgcgatc gctgccaaaa aggtaagctg acgatagctg acgagattcg atgcccgacg
     2461 agcaaatgag cgccacggct gttcgcgtaa gctcgaaacg acggagattc gatagagcca
     2521 gagtcgagtt taaacgattc gggatcttaa ggataaattt ctggtaagga ggacacgtat
     2581 ggaagtgggc aagttgggga agccgtatcc gttgctgaat ctggcatatg tgggagtata
     2641 agacgcgcag cgtcgcatca ggcatttttt tctgcgccaa tgcacctaga atcggttacc
     2701 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     2761 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     2821 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     2881 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     2941 tcaactgttg ccgggaagct agagtaagta gttcgccagt taatagttta cgcaaggttg
     3001 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3061 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3121 gctccttcgg tcctccaatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3181 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3241 ctggtgaata ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3301 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3361 ttggaaagcg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3421 cgatgtaacc cactcgcgca cccaactgat cttcagcatc ttttactttc accagcgttt
     3481 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     3541 aatgttgaat actcatgcga aagcctcctg ctgatcctgt ctcttgatag atgtttgcaa
     3601 aagctaggac tcgattatac gttgaccgat cagactatca atatctaggt ttaaacagat
     3661 cggtacgctg ccgtgaccag gcttgagtac aatatctgac gattggactt cctgggtccc
     3721 aagctggatt ccgactggct gactgccaag caacgatatg gatcgtcgag actcgcactg
     3781 agaggatcgt tctcggtaca ccaggcgcgc cccgatttcg atattcctcg gatctcggat
     3841 agatcgggat tgaggatgta ggttgaactt ctctcgggtg atcccgatct cggatattcg
     3901 ggattggagt aatacctggc gataggcgct gccaagcaat agccaaggcg caaggccggc
     3961 caagagactt gcgatagttc gttcgagagg tagcctgcaa gatggatcgg acgaaaggtt
     4021 tgaagatttt ccgacgaggg ccttggagag ttcggaaatt taaatgcaac tatggatgaa
     4081 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4141 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4201 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4261 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4321 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4381 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4441 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4501 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4561 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4621 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4681 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4741 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4801 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4861 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4921 ctggcctttt gctggccttt tgctcacatg atttaaatga aactcgtaat cggagaacgg
     4981 gtgcaatagc aaggaccaga tcgaacttga ctgacgcaat tctgagcgta gtcgaaaccc
     5041 agacaagtcg atctcgccaa gtagctgact tgactggtta attaagtttc gatagcccaa
     5101 ggtaaccaat gagattaggg agggaagaaa gtttgatgaa ctacttctga cccctagtgg
     5161 tgtccagaaa agaccattaa aggaatgtct tgatcgaaat cgagagatcg cctgcaggat
     5221 agctgacgac cttaaggata aatttctggt aaggaggaca cgtatggaag tgggcaagtt
     5281 ggggaagccg tatccgttgc tgaatctggc atatgtggga gtataagacg cgcagcgtcg
     5341 catcaggcat ttttttctgc gccaatgccg gtagatagca atccggcccg aggggcacaa
     5401 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctacgatac cgatagagat
     5461 gggccaatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     5521 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     5581 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5641 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt gagctgaagg
     5701 tacgctgtat ctcagtcagt caagctagct taactgtcca tgtgctgcct ctcgaactta
     5761 gctgccgcgg tttctttcga ctgaaagtaa aggttttctg tgtaatgtaa tttgactcct
     5821 ttgagcactg gctcagagtc gtcttcatca aatttgcagc aggatccaca agaacaacag
     5881 cccttgagac aactacagca actggtcata cagcaaagca taattgtcac cattactatg
     5941 gcaatcaagc cagctataaa acctagccaa atgtaccatg gccattttat atactgctca
     6001 tactttccaa gttcttggag atcgatgaga gattcattta aattcttggc aacctcattg
     6061 aggcggtcaa tttctttttg aatgtttaca actgaagcat taatgccaga gatgtcacct
     6121 aaatcaacat ctggtgatgt atgattctta aaatatttat ctaactcctc cttgaatgag
     6181 tctaattcag gttgcaaagg atcataaact gtgttgttga caattcctat tacaacatca
     6241 cagttaccag acacaaatgt gttgtctgta gtaatgattt gtggttcata aaaattcctt
     6301 tgtgttacaa accagtgtgt gccatttgaa acaaagacac cttcacgagg aaagtgtgct
     6361 tttccatcat gacaaatggc aggagcagtt gtgaagttct tttcttgtgc agggacataa
     6421 gtcacatgca agaagactac accatgaggt gctgactgag ggaaggacat aagatgatag
     6481 ccctttccac aaaaatcaac tctttttgat tgtccaagta cacactctga cattttagta
     6541 gcagcaagat tagcagaagc tctgatttct gcagctctaa ttaattgttg agtcacatat
     6601 gtctgcaaac tttgaagtct gcctgtgatc aacctatcaa tttgcacttc agcctcaact
     6661 ttgtcaagac gtgaaaggat atcatttaaa acacttgaaa ttgcaccaaa attggagcta
     6721 agttgtttaa caagcgtgtt taaagcttgt gcattttggt tgaccacatc ttgaagtttt
     6781 ccaagtgcac ttgctgtgga agaaagtgag tcttgaattt tgccaatagc actattaaat
     6841 tggttggcaa tcaatttttg gttctcatag agaacattct gtgtaactcc aataccatta
     6901 aacctataag ccatttgcat agcaaatggt atttgtaatg cagcacctgc accaaaggtc
     6961 caaccagaag tgattgtacc cgctaacagt gcagaagtgt attgagcaat catttcatct
     7021 gtgagcaaag gtggcaaaac agtaaggccg ttaaactttt gtgcacaaat gaggtctcta
     7081 gcagcaatat caccaaggca atcaccatat tgtttgatga agccagcatc tgcaagtgtc
     7141 actttgttga aaagtagatc ttcaataaat gacctcttgc ttggttttga tggatctggt
     7201 aatatttgtg aaaaattaaa accaccaaaa tctttaattg gtggtgtttt gtaaatttgt
     7261 ttgacttgtg caaaaacttc ttgggtgttt ttgtcttgtt caacagctat tccagttaaa
     7321 gcacggttta attgtgtaca aaaactgcca tattgcaaca aaagattgct gcattcagtt
     7381 gaatcaccac aaatgtacat tgtacaatct actgatgtct tggtcataga cactggtaga
     7441 atttctgtgg taacactaat agtaaaattt gtgggtatgg caatagagtt attagagtaa
     7501 gcaactgaat tttctgcacc aagtgacata gtgtaggcaa tgatggattg actagctaca
     7561 ctacgtgccc gccgaggaga attagtctga gtctgataac tagcgcatat acctgcacca
     7621 atgggtatgt cacactcata tgagttgttg acatgttcag cccctattaa acagcctgca
     7681 cgtgtttgaa aaacattaga acctgtagaa taaacacgcc aagtaggagt aagttgatct
     7741 gcatgaatag caacagggac ttctgtgcag ttaacatcct gataaagaac agcaacctgg
     7801 ttagaagtat ttgttcctgg tgttataaca ctgacaccac caaaagaaca tggtgtaatg
     7861 tcaagaatct caagtgtctg tggatcacgg acagcatcag tagtgtcagc aatgtctctg
     7921 ccaaattgtt ggaaaggcag aaactttttg ttagactcag taagaacacc tgtgcctgtt
     7981 aaaccattga agttgaaatt gacacatttg tttttaacca aattagtaga ctttttaggt
     8041 ccacaaacag ttgctggtgc atgtagaagt tca
//
