LOCUS       sequence-297356-        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-m4133729
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 caccatgttt gtttttcttg ttttattgcc actagtctct agtcagtgtg ttaatcttac
       61 aaccagaact caattacccc ctgcatacac taattctttc acacgtggtg tttattaccc
      121 tgacaaagtt ttcagatcct cagttttaca ttcaactcag gacttgttct tacctttctt
      181 ttccaatgtt acttggttcc atgctataca tgtctctggg accaatggta ctaagaggtt
      241 tgataaccct gtcctaccat ttaatgatgg tgtttatttt gcttccactg agaagtctaa
      301 cataataaga ggctggattt ttggtactac tttagattcg aagacccagt ccctacttat
      361 tgttaataac gctactaatg ttgttattaa agtctgtgaa tttcaatttt gtaatgatcc
      421 atttttgggt gtttattacc acaaaaacaa caaaagttgg atggaaagtg agttcagagt
      481 ttattctagt gcgaataatt gcacttttga atatgtctct cagccttttc ttatggacct
      541 tgaaggaaaa cagggtaatt tcaaaaatct tagggaattt gtgtttaaga atattgatgg
      601 ttattttaaa atatattcta agcacacgcc tattaattta gtgcgtgatc tccctcaggg
      661 tttttcggct ttagaaccat tggtagattt gccaataggt attaacatca ctaggtttca
      721 aactttactt gctttacata gaagttattt gactcctggt gattcttctt caggttggac
      781 agctggtgct gcagcttatt atgtgggtta tcttcaacct aggacttttc tattaaaata
      841 taatgaaaat ggaaccatta cagatgctgt agactgtgca cttgaccctc tctcagaaac
      901 aaagtgtacg ttgaaatcct tcactgtaga aaaaggaatc tatcaaactt ctaactttag
      961 agtccaacca acagaatcta ttgttagatt tcctaatatt acaaacttgt gcccttttgg
     1021 tgaagttttt aacgccacca gatttgcatc tgtttatgct tggaacagga agagaatcag
     1081 caactgtgtt gctgattatt ctgtcctata taattccgca tcattttcca cttttaagtg
     1141 ttatggagtg tctcctacta aattaaatga tctctgcttt actaatgtct atgcagattc
     1201 atttgtaatt agaggtgatg aagtcagaca aatcgctcca gggcaaactg gaaagattgc
     1261 tgattataat tataaattac cagatgattt tacaggctgc gttatagctt ggaattctaa
     1321 caatcttgat tctaaggttg gtggtaatta taattacctg tatagattgt ttaggaagtc
     1381 taatctcaaa ccttttgaga gagatatttc aactgaaatc tatcaggccg gtagcacacc
     1441 ttgtaatggt gttgaaggtt ttaattgtta ctttccttta caatcatatg gtttccaacc
     1501 cactaatggt gttggttacc aaccatacag agtagtagta ctttcttttg aacttctaca
     1561 tgcaccagca actgtttgtg gacctaaaaa gtctactaat ttggttaaaa acaaatgtgt
     1621 caatttcaac ttcaatggtt taacaggcac aggtgttctt actgagtcta acaaaaagtt
     1681 tctgcctttc caacaatttg gcagagacat tgctgacact actgatgctg tccgtgatcc
     1741 acagacactt gagattcttg acattacacc atgttctttt ggtggtgtca gtgttataac
     1801 accaggaaca aatacttcta accaggttgc tgttctttat caggatgtta actgcacaga
     1861 agtccctgtt gctattcatg cagatcaact tactcctact tggcgtgttt attctacagg
     1921 ttctaatgtt tttcaaacac gtgcaggctg tttaataggg gctgaacatg tcaacaactc
     1981 atatgagtgt gacataccca ttggtgcagg tatatgcgct agttatcaga ctcagactaa
     2041 ttctcctcgg cgggcacgta gtgtagctag tcaatccatc attgcctaca ctatgtcact
     2101 tggtgcagaa aattcagttg cttactctaa taactctatt gccataccca caaattttac
     2161 tattagtgtt accacagaaa ttctaccagt gtctatgacc aagacatcag tagattgtac
     2221 aatgtacatt tgtggtgatt caactgaatg cagcaatctt ttgttgcaat atggcagttt
     2281 ttgtacacaa ttaaaccgtg ctttaactgg aatagctgtt gaacaagaca aaaacaccca
     2341 agaagttttt gcacaagtca aacaaattta caaaacacca ccaattaaag attttggtgg
     2401 ttttaatttt tcacaaatat taccagatcc atcaaaacca agcaagaggt catttattga
     2461 agatctactt ttcaacaaag tgacacttgc agatgctggc ttcatcaaac aatatggtga
     2521 ttgccttggt gatattgctg ctagagacct catttgtgca caaaagttta acggccttac
     2581 tgttttgcca cctttgctca cagatgaaat gattgctcaa tacacttctg cactgttagc
     2641 gggtacaatc acttctggtt ggacctttgg tgcaggtgct gcattacaaa taccatttgc
     2701 tatgcaaatg gcttataggt ttaatggtat tggagttaca cagaatgttc tctatgagaa
     2761 ccaaaaattg attgccaacc aatttaatag tgctattggc aaaattcaag actcactttc
     2821 ttccacagca agtgcacttg gaaaacttca agatgtggtc aaccaaaatg cacaagcttt
     2881 aaacacgctt gttaaacaac ttagctccaa ttttggtgca atttcaagtg ttttaaatga
     2941 tatcctttca cgtcttgaca aagttgaggc tgaagtgcaa attgataggt tgatcacagg
     3001 cagacttcaa agtttgcaga catatgtgac tcaacaatta attagagctg cagaaatcag
     3061 agcttctgct aatcttgctg ctactaaaat gtcagagtgt gtacttggac aatcaaaaag
     3121 agttgatttt tgtggaaagg gctatcatct tatgtccttc cctcagtcag cacctcatgg
     3181 tgtagtcttc ttgcatgtga cttatgtccc tgcacaagaa aagaacttca caactgctcc
     3241 tgccatttgt catgatggaa aagcacactt tcctcgtgaa ggtgtctttg tttcaaatgg
     3301 cacacactgg tttgtaacac aaaggaattt ttatgaacca caaatcatta ctacagacaa
     3361 cacatttgtg tctggtaact gtgatgttgt aataggaatt gtcaacaaca cagtttatga
     3421 tcctttgcaa cctgaattag actcattcaa ggaggagtta gataaatatt ttaagaatca
     3481 tacatcacca gatgttgatt taggtgacat ctctggcatt aatgcttcag ttgtaaacat
     3541 tcaaaaagaa attgaccgcc tcaatgaggt tgccaagaat ttaaatgaat ctctcatcga
     3601 tctccaagaa cttggaaagt atgagcagta tataaaatgg ccatggtaca tttggctagg
     3661 ttttatagct ggcttgattg ccatagtaat ggtgacaatt atgctttgct gtatgaccag
     3721 ttgctgtagt tgtctcaagg gctgttgttc ttgtggatcc tgctgcaaat ttgatgaaga
     3781 cgactctgag ccagtgctca aaggagtcaa attacattac 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
//
