LOCUS       sequence-295690-        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"
     CDS             3824..3844
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3878..3955
                     /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             3983..4003
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    4452..4573
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4463..4482)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4517..4536
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4688..4734
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4764..4900
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4945..5016
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(5242..5830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5322..5341)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(6576..6644,6645..7436)
                     /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(6794..6813)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7449..7585
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7737..7808
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7813..7859)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7877..7925
                     /note="synthetic polyadenylation signal"
     enhancer        8006..8309
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8311..8513
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8464..8484
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgttc gtctttcttg tgttactccc cctggtttcg tcccagtgcg tgaacttgac
       61 tacgcgaacc caactgccgc cagcatatac aaatagtttc acccgtggag tatactaccc
      121 tgataaagtg tttagatcaa gcgtactaca ctctactcag gacctgtttc tccccttctt
      181 ctccaatgtc acatggtttc atgcgatcca cgttagcggc accaacggga caaagcgctt
      241 tgacaaccct gtgctgccat tcaatgatgg tgtgtatttt gcttctaccg agaagagcaa
      301 cataattcgg ggctggatct tcggaactac cctcgacagt aaaactcagt cattgctgat
      361 tgtgaataac gccacaaatg tcgttatcaa ggtctgtgaa tttcaattct gcaacgatcc
      421 attccttggg gtgtattacc ataaaaataa taagtcctgg atggagagcg aatttagggt
      481 atactcgtct gccaacaatt gtactttcga gtatgtgtct cagcccttct taatggatct
      541 ggaaggtaaa cagggcaact ttaagaacct ccgcgagttt gtattcaaaa atatagacgg
      601 atactttaag atttattcca agcacacacc aatcaatttg gtgcgggatc tcccgcaagg
      661 gttcagcgct ctggaacctc tagtggacct ccccatcggc attaacatca caagattcca
      721 gacgctgtta gccctgcata gatcatacct tactcccggt gacagtagtt ctgggtggac
      781 cgcaggcgcg gctgcatatt acgtgggata tttgcagcct cggacgtttc tgctgaaata
      841 caacgagaac ggcaccatta cagatgccgt ggattgcgcc cttgaccccc taagcgaaac
      901 taagtgtacg cttaaaagct tcaccgtcga gaagggaatt taccagactt ccaactttag
      961 ggtccagccg acagaatcaa tagttaggtt cccaaatatc accaatttgt gcccttttgg
     1021 ggaagtcttc aatgctacac gatttgcttc ggtttatgcc tggaacagga agcgtatctc
     1081 aaattgcgtc gcggactaca gcgtgctcta taatagtgca agtttctcga cctttaaatg
     1141 ttatggcgtg tccccgacca aactaaatga cctctgtttt acaaacgttt acgccgattc
     1201 ttttgtgatt cgcggtgatg aggtacgcca aatcgcaccg ggccagactg gcaagattgc
     1261 cgattacaac tacaagttgc ctgacgactt caccggatgt gttatagcct ggaatagcaa
     1321 taatctggat tctaaggtcg gaggtaacta taactacctg tatagactgt tccgtaagtc
     1381 aaacctgaaa ccctttgagc gggacatctc cacagagatt taccaagcag ggtctacccc
     1441 ctgcaatggg gttgaaggat tcaattgcta ctttccatta cagagctacg ggttccagcc
     1501 tacgaacggg gttggttacc agccttatag ggtagtggtg ctgtcgttcg aactgttgca
     1561 cgctccagct acagtgtgcg ggcctaaaaa atcaactaac ctggttaaaa ataagtgtgt
     1621 taattttaac ttcaatggct taaccggaac tggagtgctc acggagtcaa acaagaaatt
     1681 tctaccgttt cagcaattcg ggagggatat cgcagacact acggacgcgg tcagagatcc
     1741 acagaccctt gagatccttg acataacacc ctgctctttc ggaggtgtct ccgtaattac
     1801 accaggcact aatacctcta atcaggtggc cgtgctttat caggatgtta actgtacaga
     1861 ggtcccggtg gctatccatg ccgaccaatt gacacctacg tggcgcgtct attctaccgg
     1921 ctccaacgtc ttccagaccc gggctggatg cttgataggc gcagaacacg taaacaactc
     1981 ttacgaatgt gatattccca ttggagccgg gatctgcgca agctaccaga cccaaacaaa
     2041 cagtcccaga cgggctcgta gcgtggcctc tcagagcata atagcctata ccatgtccct
     2101 tggcgcggag aatagtgtgg catactcaaa taatagcatc gctatcccca ctaactttac
     2161 tatttccgtg acgactgaga ttctgccagt gtctatgact aagacatccg tggattgcac
     2221 aatgtatatt tgtggggatt ccacagagtg tagcaacttg ttgctgcaat atggatcatt
     2281 ctgtacgcag ctgaatcgcg ccctcaccgg catcgctgtg gaacaggaca agaacactca
     2341 agaggtcttt gctcaggtga agcaaatcta caagacccca cctataaaag acttcggggg
     2401 ttttaacttc tcgcagattc tccccgaccc cagcaagcct tcaaaaagga gctttattga
     2461 ggacctgttg ttcaacaagg ttactttagc cgatgccggt tttatcaaac agtacggcga
     2521 ctgtcttggc gatatagcag cccgggatct gatttgcgca caaaaattca atggccttac
     2581 cgtactgcca ccccttctca ccgacgaaat gatcgcgcag tacacctccg cactgctggc
     2641 tgggaccatc acttcgggat ggacctttgg ggctggtgct gctttgcaaa ttcccttcgc
     2701 catgcagatg gcctatcgat tcaatggcat aggggtaaca cagaacgtat tgtatgaaaa
     2761 tcaaaagcta atcgctaacc agtttaattc agccattggt aagatccagg attcactctc
     2821 cagtacagcc tctgccctgg gaaagctcca ggacgtggtc aaccagaacg cgcaggcatt
     2881 gaatacactc gtcaaacagc tgagtagcaa tttcggcgca atcagtagtg tcttaaacga
     2941 tatactgagt aggcttgata aagtagaggc cgaggtccag attgacagac taatcacggg
     3001 gcgtcttcag agtttacaga cctatgtgac ccaacagctg atccgagcag cggaaattcg
     3061 agcaagcgcg aacctggccg ctactaaaat gagtgagtgc gtgctgggcc aaagtaaacg
     3121 agtcgacttt tgcggcaaag ggtatcatct catgtctttc cctcagtccg cccctcacgg
     3181 tgttgttttc ctccacgtga catacgttcc cgcgcaggaa aagaatttta ctacggcacc
     3241 cgctatttgc catgacggaa aggctcattt tccacgggag ggagtgttcg tttcaaacgg
     3301 gacacactgg tttgtaactc agcggaactt ttatgagcct caaatcatca cgacagacaa
     3361 taccttcgtg agtggcaact gcgatgtagt cataggcatt gtgaataaca ctgtttatga
     3421 tccacttcag ccggaactcg acagcttcaa agaggaattg gataagtact ttaaaaatca
     3481 taccagtcca gacgtagatc tcggcgatat atcgggcatt aacgccagtg tggtcaatat
     3541 acaaaaggag attgacagac tgaatgaagt agctaagaac ctcaacgaat ctctgattga
     3601 cctgcaagag ctagggaagt acgagcagta tatcaaatgg ccatggtaca tatggctggg
     3661 ttttatcgcg ggtcttattg ccatagtcat ggtgacgatt atgttgtgct gtatgacctc
     3721 ctgctgttct tgtttgaaag gctgctgctc ctgtggtagc tgttgcaaat tcgatgaaga
     3781 tgactctgaa cccgtgttga aaggcgttaa gctacactac acagaaaacc tttactttca
     3841 gtcgggacgg cgaatcccgg ggctgataaa tccctggaaa aggagatgga agaagaactt
     3901 tattgcagtg tctgcggcta accggttcaa aaagatcagt tcaagcggcg ccctagatta
     3961 cgacattcct acgaccgcct ccgagaatct ctatttccag ggtgaatttg gcttggcaca
     4021 acatgacgag gccgtcgata acaagtttaa taaagaacag cagaacgctt tctatgagat
     4081 ccttcactta ccaaatctga acgaagagca acgcaacgct ttcattcaga gcctgaaaga
     4141 cgatccctcg cagtctgcaa atctcctggc cgaagccaag aagctgaatg acgcacaggc
     4201 tccaaaagtt gataacaagt ttaacaagga gcagcagaat gctttctacg agatattgca
     4261 ccttcctaat ctgaatgagg aacaaagaaa tgcatttatc cagtccctaa aagacgatcc
     4321 atcacagagt gccaacctgc ttgcggaagc caaaaagctc aatgatgccc aggccccgaa
     4381 agtagacgcg aaccatcaat aagctagctt gactgactga gatacagcgt accttcagct
     4441 cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     4501 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     4561 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     4621 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc
     4681 ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc
     4741 tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc
     4801 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
     4861 acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag
     4921 gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg
     4981 gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat
     5041 cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga
     5101 ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc
     5161 gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5221 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5281 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5341 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5401 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5461 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5521 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5581 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5641 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5701 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5761 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5821 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5881 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5941 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6001 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6061 agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg
     6121 tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt
     6181 gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc
     6241 gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat
     6301 ccgaggaata tcgaaatcgg ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag
     6361 tctcgacgat ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag
     6421 gaagtccaat cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta
     6481 aacctagata ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca
     6541 tctatcaaga gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg
     6601 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     6661 tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc
     6721 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca
     6781 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     6841 tcggtcgccg catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa
     6901 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     6961 ataacactgc ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt
     7021 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     7081 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc
     7141 gtaaactatt aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga
     7201 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     7261 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     7321 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     7381 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga
     7441 ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
     7501 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
     7561 ctccttacca gaaatttatc cttaagatcc cgaatcgttt aaactcgact ctggctctat
     7621 cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca
     7681 tcgaatctcg tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc
     7741 ttggaccatt agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc
     7801 tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat
     7861 caatcgttgc gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt
     7921 ttattatcta actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag
     7981 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     8041 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     8101 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     8161 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     8221 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     8281 tctacgtatt agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc
     8341 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     8401 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     8461 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
     8521 tgaaccgtca gatcagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc
     8581 cgc
//
