LOCUS       sequence-300225-        8583 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 8583)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8583)
  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-m4133712
FEATURES             Location/Qualifiers
     source          1..8583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4098..4118
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4152..4229
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             4257..4277
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    4726..4847
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4737..4756)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4791..4810
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4962..5008
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5038..5174
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5219..5290
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(5516..6104)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5596..5615)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(6850..6918,6919..7710)
                     /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(7068..7087)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7723..7859
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8011..8082
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8087..8133)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    8151..8199
                     /note="synthetic polyadenylation signal"
     enhancer        8280..8583
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ctgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc
      241 ctaccatcca ctcgacacac ccgccagcgg ccgccaccat gttcgtcttt cttgtgttac
      301 tccccctggt ttcgtcccag tgcgtgaact tgactacgcg aacccaactg ccgccagcat
      361 atacaaatag tttcacccgt ggagtatact accctgataa agtgtttaga tcaagcgtac
      421 tacactctac tcaggacctg tttctcccct tcttctccaa tgtcacatgg tttcatgcga
      481 tccacgttag cggcaccaac gggacaaagc gctttgacaa ccctgtgctg ccattcaatg
      541 atggtgtgta ttttgcttct accgagaaga gcaacataat tcggggctgg atcttcggaa
      601 ctaccctcga cagtaaaact cagtcattgc tgattgtgaa taacgccaca aatgtcgtta
      661 tcaaggtctg tgaatttcaa ttctgcaacg atccattcct tggggtgtat taccataaaa
      721 ataataagtc ctggatggag agcgaattta gggtatactc gtctgccaac aattgtactt
      781 tcgagtatgt gtctcagccc ttcttaatgg atctggaagg taaacagggc aactttaaga
      841 acctccgcga gtttgtattc aaaaatatag acggatactt taagatttat tccaagcaca
      901 caccaatcaa tttggtgcgg gatctcccgc aagggttcag cgctctggaa cctctagtgg
      961 acctccccat cggcattaac atcacaagat tccagacgct gttagccctg catagatcat
     1021 accttactcc cggtgacagt agttctgggt ggaccgcagg cgcggctgca tattacgtgg
     1081 gatatttgca gcctcggacg tttctgctga aatacaacga gaacggcacc attacagatg
     1141 ccgtggattg cgcccttgac cccctaagcg aaactaagtg tacgcttaaa agcttcaccg
     1201 tcgagaaggg aatttaccag acttccaact ttagggtcca gccgacagaa tcaatagtta
     1261 ggttcccaaa tatcaccaat ttgtgccctt ttggggaagt cttcaatgct acacgatttg
     1321 cttcggttta tgcctggaac aggaagcgta tctcaaattg cgtcgcggac tacagcgtgc
     1381 tctataatag tgcaagtttc tcgaccttta aatgttatgg cgtgtccccg accaaactaa
     1441 atgacctctg ttttacaaac gtttacgccg attcttttgt gattcgcggt gatgaggtac
     1501 gccaaatcgc accgggccag actggcaaga ttgccgatta caactacaag ttgcctgacg
     1561 acttcaccgg atgtgttata gcctggaata gcaataatct ggattctaag gtcggaggta
     1621 actataacta cctgtataga ctgttccgta agtcaaacct gaaacccttt gagcgggaca
     1681 tctccacaga gatttaccaa gcagggtcta ccccctgcaa tggggttgaa ggattcaatt
     1741 gctactttcc attacagagc tacgggttcc agcctacgaa cggggttggt taccagcctt
     1801 atagggtagt ggtgctgtcg ttcgaactgt tgcacgctcc agctacagtg tgcgggccta
     1861 aaaaatcaac taacctggtt aaaaataagt gtgttaattt taacttcaat ggcttaaccg
     1921 gaactggagt gctcacggag tcaaacaaga aatttctacc gtttcagcaa ttcgggaggg
     1981 atatcgcaga cactacggac gcggtcagag atccacagac ccttgagatc cttgacataa
     2041 caccctgctc tttcggaggt gtctccgtaa ttacaccagg cactaatacc tctaatcagg
     2101 tggccgtgct ttatcaggat gttaactgta cagaggtccc ggtggctatc catgccgacc
     2161 aattgacacc tacgtggcgc gtctattcta ccggctccaa cgtcttccag acccgggctg
     2221 gatgcttgat aggcgcagaa cacgtaaaca actcttacga atgtgatatt cccattggag
     2281 ccgggatctg cgcaagctac cagacccaaa caaacagtcc cagacgggct cgtagcgtgg
     2341 cctctcagag cataatagcc tataccatgt cccttggcgc ggagaatagt gtggcatact
     2401 caaataatag catcgctatc cccactaact ttactatttc cgtgacgact gagattctgc
     2461 cagtgtctat gactaagaca tccgtggatt gcacaatgta tatttgtggg gattccacag
     2521 agtgtagcaa cttgttgctg caatatggat cattctgtac gcagctgaat cgcgccctca
     2581 ccggcatcgc tgtggaacag gacaagaaca ctcaagaggt ctttgctcag gtgaagcaaa
     2641 tctacaagac cccacctata aaagacttcg ggggttttaa cttctcgcag attctccccg
     2701 accccagcaa gccttcaaaa aggagcttta ttgaggacct gttgttcaac aaggttactt
     2761 tagccgatgc cggttttatc aaacagtacg gcgactgtct tggcgatata gcagcccggg
     2821 atctgatttg cgcacaaaaa ttcaatggcc ttaccgtact gccacccctt ctcaccgacg
     2881 aaatgatcgc gcagtacacc tccgcactgc tggctgggac catcacttcg ggatggacct
     2941 ttggggctgg tgctgctttg caaattccct tcgccatgca gatggcctat cgattcaatg
     3001 gcataggggt aacacagaac gtattgtatg aaaatcaaaa gctaatcgct aaccagttta
     3061 attcagccat tggtaagatc caggattcac tctccagtac agcctctgcc ctgggaaagc
     3121 tccaggacgt ggtcaaccag aacgcgcagg cattgaatac actcgtcaaa cagctgagta
     3181 gcaatttcgg cgcaatcagt agtgtcttaa acgatatact gagtaggctt gataaagtag
     3241 aggccgaggt ccagattgac agactaatca cggggcgtct tcagagttta cagacctatg
     3301 tgacccaaca gctgatccga gcagcggaaa ttcgagcaag cgcgaacctg gccgctacta
     3361 aaatgagtga gtgcgtgctg ggccaaagta aacgagtcga cttttgcggc aaagggtatc
     3421 atctcatgtc tttccctcag tccgcccctc acggtgttgt tttcctccac gtgacatacg
     3481 ttcccgcgca ggaaaagaat tttactacgg cacccgctat ttgccatgac ggaaaggctc
     3541 attttccacg ggagggagtg ttcgtttcaa acgggacaca ctggtttgta actcagcgga
     3601 acttttatga gcctcaaatc atcacgacag acaatacctt cgtgagtggc aactgcgatg
     3661 tagtcatagg cattgtgaat aacactgttt atgatccact tcagccggaa ctcgacagct
     3721 tcaaagagga attggataag tactttaaaa atcataccag tccagacgta gatctcggcg
     3781 atatatcggg cattaacgcc agtgtggtca atatacaaaa ggagattgac agactgaatg
     3841 aagtagctaa gaacctcaac gaatctctga ttgacctgca agagctaggg aagtacgagc
     3901 agtatatcaa atggccatgg tacatatggc tgggttttat cgcgggtctt attgccatag
     3961 tcatggtgac gattatgttg tgctgtatga cctcctgctg ttcttgtttg aaaggctgct
     4021 gctcctgtgg tagctgttgc aaattcgatg aagatgactc tgaacccgtg ttgaaaggcg
     4081 ttaagctaca ctacacagaa aacctttact ttcagtcggg acggcgaatc ccggggctga
     4141 taaatccctg gaaaaggaga tggaagaaga actttattgc agtgtctgcg gctaaccggt
     4201 tcaaaaagat cagttcaagc ggcgccctag attacgacat tcctacgacc gcctccgaga
     4261 atctctattt ccagggtgaa tttggcttgg cacaacatga cgaggccgtc gataacaagt
     4321 ttaataaaga acagcagaac gctttctatg agatccttca cttaccaaat ctgaacgaag
     4381 agcaacgcaa cgctttcatt cagagcctga aagacgatcc ctcgcagtct gcaaatctcc
     4441 tggccgaagc caagaagctg aatgacgcac aggctccaaa agttgataac aagtttaaca
     4501 aggagcagca gaatgctttc tacgagatat tgcaccttcc taatctgaat gaggaacaaa
     4561 gaaatgcatt tatccagtcc ctaaaagacg atccatcaca gagtgccaac ctgcttgcgg
     4621 aagccaaaaa gctcaatgat gcccaggccc cgaaagtaga cgcgaaccat caataagcta
     4681 gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg
     4741 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     4801 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     4861 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     4921 acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc
     4981 ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca
     5041 ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag
     5101 attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag
     5161 aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca
     5221 ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct
     5281 tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca
     5341 gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag
     5401 ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt
     5461 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5521 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5581 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5641 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5701 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5761 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5821 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5881 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5941 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6001 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6061 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6121 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6181 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6241 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6301 ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc
     6361 caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct
     6421 cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc
     6481 ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt
     6541 caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg
     6601 cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg
     6661 cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact
     6721 caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg
     6781 tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga
     6841 ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6901 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6961 ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     7021 tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct atgtggcgcg
     7081 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     7141 aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg catgacagta
     7201 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     7261 acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     7321 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     7381 accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg cgaactactt
     7441 actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt tgcaggacca
     7501 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     7561 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     7621 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7681 ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc gcagaaaaaa
     7741 atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata
     7801 cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag
     7861 atcccgaatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg
     7921 cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct
     7981 tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct
     8041 tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca
     8101 agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa
     8161 accaacacac atccatcttc gatggatagc gattttatta tctaactgct gatcgagtgt
     8221 agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     8281 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     8341 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     8401 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     8461 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     8521 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     8581 atg
//
