LOCUS       sequence-313661-        8097 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 8097)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8097)
  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-m4133151
FEATURES             Location/Qualifiers
     source          1..8097
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4402..4422
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4426..4491
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    4544..4665
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4555..4574)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4609..4628
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4780..4826
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4856..4992
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5037..5108
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(5334..5922)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5414..5433)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(6668..6736,6737..7528)
                     /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(6886..6905)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7541..7677
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7829..7900
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7905..7951)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7969..8017
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgttcgt ctttcttgtg
      601 ttactccccc tggtttcgtc ccagtgcgtg aacttgacta cgcgaaccca actgccgcca
      661 gcatatacaa atagtttcac ccgtggagta tactaccctg ataaagtgtt tagatcaagc
      721 gtactacact ctactcagga cctgtttctc cccttcttct ccaatgtcac atggtttcat
      781 gcgatccacg ttagcggcac caacgggaca aagcgctttg acaaccctgt gctgccattc
      841 aatgatggtg tgtattttgc ttctaccgag aagagcaaca taattcgggg ctggatcttc
      901 ggaactaccc tcgacagtaa aactcagtca ttgctgattg tgaataacgc cacaaatgtc
      961 gttatcaagg tctgtgaatt tcaattctgc aacgatccat tccttggggt gtattaccat
     1021 aaaaataata agtcctggat ggagagcgaa tttagggtat actcgtctgc caacaattgt
     1081 actttcgagt atgtgtctca gcccttctta atggatctgg aaggtaaaca gggcaacttt
     1141 aagaacctcc gcgagtttgt attcaaaaat atagacggat actttaagat ttattccaag
     1201 cacacaccaa tcaatttggt gcgggatctc ccgcaagggt tcagcgctct ggaacctcta
     1261 gtggacctcc ccatcggcat taacatcaca agattccaga cgctgttagc cctgcataga
     1321 tcatacctta ctcccggtga cagtagttct gggtggaccg caggcgcggc tgcatattac
     1381 gtgggatatt tgcagcctcg gacgtttctg ctgaaataca acgagaacgg caccattaca
     1441 gatgccgtgg attgcgccct tgacccccta agcgaaacta agtgtacgct taaaagcttc
     1501 accgtcgaga agggaattta ccagacttcc aactttaggg tccagccgac agaatcaata
     1561 gttaggttcc caaatatcac caatttgtgc ccttttgggg aagtcttcaa tgctacacga
     1621 tttgcttcgg tttatgcctg gaacaggaag cgtatctcaa attgcgtcgc ggactacagc
     1681 gtgctctata atagtgcaag tttctcgacc tttaaatgtt atggcgtgtc cccgaccaaa
     1741 ctaaatgacc tctgttttac aaacgtttac gccgattctt ttgtgattcg cggtgatgag
     1801 gtacgccaaa tcgcaccggg ccagactggc aagattgccg attacaacta caagttgcct
     1861 gacgacttca ccggatgtgt tatagcctgg aatagcaata atctggattc taaggtcgga
     1921 ggtaactata actacctgta tagactgttc cgtaagtcaa acctgaaacc ctttgagcgg
     1981 gacatctcca cagagattta ccaagcaggg tctaccccct gcaatggggt tgaaggattc
     2041 aattgctact ttccattaca gagctacggg ttccagccta cgaacggggt tggttaccag
     2101 ccttataggg tagtggtgct gtcgttcgaa ctgttgcacg ctccagctac agtgtgcggg
     2161 cctaaaaaat caactaacct ggttaaaaat aagtgtgtta attttaactt caatggctta
     2221 accggaactg gagtgctcac ggagtcaaac aagaaatttc taccgtttca gcaattcggg
     2281 agggatatcg cagacactac ggacgcggtc agagatccac agacccttga gatccttgac
     2341 ataacaccct gctctttcgg aggtgtctcc gtaattacac caggcactaa tacctctaat
     2401 caggtggccg tgctttatca ggatgttaac tgtacagagg tcccggtggc tatccatgcc
     2461 gaccaattga cacctacgtg gcgcgtctat tctaccggct ccaacgtctt ccagacccgg
     2521 gctggatgct tgataggcgc agaacacgta aacaactctt acgaatgtga tattcccatt
     2581 ggagccggga tctgcgcaag ctaccagacc caaacaaaca gtcccagacg ggctcgtagc
     2641 gtggcctctc agagcataat agcctatacc atgtcccttg gcgcggagaa tagtgtggca
     2701 tactcaaata atagcatcgc tatccccact aactttacta tttccgtgac gactgagatt
     2761 ctgccagtgt ctatgactaa gacatccgtg gattgcacaa tgtatatttg tggggattcc
     2821 acagagtgta gcaacttgtt gctgcaatat ggatcattct gtacgcagct gaatcgcgcc
     2881 ctcaccggca tcgctgtgga acaggacaag aacactcaag aggtctttgc tcaggtgaag
     2941 caaatctaca agaccccacc tataaaagac ttcgggggtt ttaacttctc gcagattctc
     3001 cccgacccca gcaagccttc aaaaaggagc tttattgagg acctgttgtt caacaaggtt
     3061 actttagccg atgccggttt tatcaaacag tacggcgact gtcttggcga tatagcagcc
     3121 cgggatctga tttgcgcaca aaaattcaat ggccttaccg tactgccacc ccttctcacc
     3181 gacgaaatga tcgcgcagta cacctccgca ctgctggctg ggaccatcac ttcgggatgg
     3241 acctttgggg ctggtgctgc tttgcaaatt cccttcgcca tgcagatggc ctatcgattc
     3301 aatggcatag gggtaacaca gaacgtattg tatgaaaatc aaaagctaat cgctaaccag
     3361 tttaattcag ccattggtaa gatccaggat tcactctcca gtacagcctc tgccctggga
     3421 aagctccagg acgtggtcaa ccagaacgcg caggcattga atacactcgt caaacagctg
     3481 agtagcaatt tcggcgcaat cagtagtgtc ttaaacgata tactgagtag gcttgataaa
     3541 gtagaggccg aggtccagat tgacagacta atcacggggc gtcttcagag tttacagacc
     3601 tatgtgaccc aacagctgat ccgagcagcg gaaattcgag caagcgcgaa cctggccgct
     3661 actaaaatga gtgagtgcgt gctgggccaa agtaaacgag tcgacttttg cggcaaaggg
     3721 tatcatctca tgtctttccc tcagtccgcc cctcacggtg ttgttttcct ccacgtgaca
     3781 tacgttcccg cgcaggaaaa gaattttact acggcacccg ctatttgcca tgacggaaag
     3841 gctcattttc cacgggaggg agtgttcgtt tcaaacggga cacactggtt tgtaactcag
     3901 cggaactttt atgagcctca aatcatcacg acagacaata ccttcgtgag tggcaactgc
     3961 gatgtagtca taggcattgt gaataacact gtttatgatc cacttcagcc ggaactcgac
     4021 agcttcaaag aggaattgga taagtacttt aaaaatcata ccagtccaga cgtagatctc
     4081 ggcgatatat cgggcattaa cgccagtgtg gtcaatatac aaaaggagat tgacagactg
     4141 aatgaagtag ctaagaacct caacgaatct ctgattgacc tgcaagagct agggaagtac
     4201 gagcagtata tcaaatggcc atggtacata tggctgggtt ttatcgcggg tcttattgcc
     4261 atagtcatgg tgacgattat gttgtgctgt atgacctcct gctgttcttg tttgaaaggc
     4321 tgctgctcct gtggtagctg ttgcaaattc gatgaagatg actctgaacc cgtgttgaaa
     4381 ggcgttaagc tacactacac agaaaacctt tactttcagt cgatggatta taaggaccac
     4441 gatggagact acaaagatca tgacattgat tacaaggacg acgatgacaa ataagctagc
     4501 ttgactgact gagatacagc gtaccttcag ctcacagaca tgataagata cattgatgag
     4561 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     4621 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     4681 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     4741 ctctacaaat gtggtattgg cccatctcta tcggtatcgt agcataaccc cttggggcct
     4801 ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg gattgctatc taccggcatt
     4861 ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat
     4921 tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa
     4981 atttatcctt aaggtcgtca gctatcctgc aggcgatctc tcgatttcga tcaagacatt
     5041 cctttaatgg tcttttctgg acaccactag gggtcagaag tagttcatca aactttcttc
     5101 cctccctaat ctcattggtt accttgggct atcgaaactt aattaaccag tcaagtcagc
     5161 tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt cagtcaagtt
     5221 cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta aatcatgtga
     5281 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5341 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5401 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5461 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5521 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5581 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5641 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5701 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5761 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5821 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5881 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5941 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6001 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6061 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6121 atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc gaactctcca
     6181 aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg
     6241 aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt ggcagcgcct
     6301 atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga gagaagttca
     6361 acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc
     6421 tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg ttgcttggca
     6481 gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat attgtactca
     6541 agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt ctgatcggtc
     6601 aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat cagcaggagg
     6661 ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6721 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6781 gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6841 tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6901 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6961 tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     7021 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     7081 aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     7141 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     7201 cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg aactacttac
     7261 tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg caggaccact
     7321 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     7381 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     7441 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     7501 aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc agaaaaaaat
     7561 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
     7621 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaagat
     7681 cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg agcttacgcg
     7741 aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat cgtcagctta
     7801 cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac aggtatcttc
     7861 ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa acccctcaag
     7921 acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca cacaaaaaac
     7981 caacacacat ccatcttcga tggatagcga ttttattatc taactgctga tcgagtgtag
     8041 ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccg
//
