LOCUS       sequence-307373-        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"
     rep_origin      complement(49..637)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(129..148)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(1383..1451,1452..2243)
                     /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(1601..1620)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      2256..2392
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2544..2615
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2620..2666)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    2684..2732
                     /note="synthetic polyadenylation signal"
     enhancer        2813..3116
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3118..3320
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3271..3291
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             7214..7234
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             7268..7345
                     /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             7373..7393
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    7842..7963
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7853..7872)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7907..7926
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      8078..8124
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      8154..8290
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8335..8406
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
ORIGIN
        1 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
       61 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
      121 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
      181 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
      241 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
      301 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
      361 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
      421 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
      481 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
      541 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
      601 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
      661 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
      721 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
      781 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
      841 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
      901 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
      961 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     1021 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     1081 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggtg
     1141 taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag
     1201 ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct
     1261 ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt
     1321 ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc
     1381 gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1441 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     1501 cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     1561 ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     1621 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     1681 tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     1741 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     1801 ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     1861 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     1921 tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag
     1981 cttcccggca acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc
     2041 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2101 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     2161 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     2221 cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     2281 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     2341 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga
     2401 atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag
     2461 ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt
     2521 ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc
     2581 tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg
     2641 tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca
     2701 cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga
     2761 tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     2821 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     2881 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     2941 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     3001 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     3061 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga
     3121 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     3181 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     3241 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     3301 aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac
     3361 catccactcg acacacccgc cagcggccgc caccatgttt gtttttcttg ttttattgcc
     3421 actagtctct agtcagtgtg ttaatcttac aaccagaact caattacccc ctgcatacac
     3481 taattctttc acacgtggtg tttattaccc tgacaaagtt ttcagatcct cagttttaca
     3541 ttcaactcag gacttgttct tacctttctt ttccaatgtt acttggttcc atgctataca
     3601 tgtctctggg accaatggta ctaagaggtt tgataaccct gtcctaccat ttaatgatgg
     3661 tgtttatttt gcttccactg agaagtctaa cataataaga ggctggattt ttggtactac
     3721 tttagattcg aagacccagt ccctacttat tgttaataac gctactaatg ttgttattaa
     3781 agtctgtgaa tttcaatttt gtaatgatcc atttttgggt gtttattacc acaaaaacaa
     3841 caaaagttgg atggaaagtg agttcagagt ttattctagt gcgaataatt gcacttttga
     3901 atatgtctct cagccttttc ttatggacct tgaaggaaaa cagggtaatt tcaaaaatct
     3961 tagggaattt gtgtttaaga atattgatgg ttattttaaa atatattcta agcacacgcc
     4021 tattaattta gtgcgtgatc tccctcaggg tttttcggct ttagaaccat tggtagattt
     4081 gccaataggt attaacatca ctaggtttca aactttactt gctttacata gaagttattt
     4141 gactcctggt gattcttctt caggttggac agctggtgct gcagcttatt atgtgggtta
     4201 tcttcaacct aggacttttc tattaaaata taatgaaaat ggaaccatta cagatgctgt
     4261 agactgtgca cttgaccctc tctcagaaac aaagtgtacg ttgaaatcct tcactgtaga
     4321 aaaaggaatc tatcaaactt ctaactttag agtccaacca acagaatcta ttgttagatt
     4381 tcctaatatt acaaacttgt gcccttttgg tgaagttttt aacgccacca gatttgcatc
     4441 tgtttatgct tggaacagga agagaatcag caactgtgtt gctgattatt ctgtcctata
     4501 taattccgca tcattttcca cttttaagtg ttatggagtg tctcctacta aattaaatga
     4561 tctctgcttt actaatgtct atgcagattc atttgtaatt agaggtgatg aagtcagaca
     4621 aatcgctcca gggcaaactg gaaagattgc tgattataat tataaattac cagatgattt
     4681 tacaggctgc gttatagctt ggaattctaa caatcttgat tctaaggttg gtggtaatta
     4741 taattacctg tatagattgt ttaggaagtc taatctcaaa ccttttgaga gagatatttc
     4801 aactgaaatc tatcaggccg gtagcacacc ttgtaatggt gttgaaggtt ttaattgtta
     4861 ctttccttta caatcatatg gtttccaacc cactaatggt gttggttacc aaccatacag
     4921 agtagtagta ctttcttttg aacttctaca tgcaccagca actgtttgtg gacctaaaaa
     4981 gtctactaat ttggttaaaa acaaatgtgt caatttcaac ttcaatggtt taacaggcac
     5041 aggtgttctt actgagtcta acaaaaagtt tctgcctttc caacaatttg gcagagacat
     5101 tgctgacact actgatgctg tccgtgatcc acagacactt gagattcttg acattacacc
     5161 atgttctttt ggtggtgtca gtgttataac accaggaaca aatacttcta accaggttgc
     5221 tgttctttat caggatgtta actgcacaga agtccctgtt gctattcatg cagatcaact
     5281 tactcctact tggcgtgttt attctacagg ttctaatgtt tttcaaacac gtgcaggctg
     5341 tttaataggg gctgaacatg tcaacaactc atatgagtgt gacataccca ttggtgcagg
     5401 tatatgcgct agttatcaga ctcagactaa ttctcctcgg cgggcacgta gtgtagctag
     5461 tcaatccatc attgcctaca ctatgtcact tggtgcagaa aattcagttg cttactctaa
     5521 taactctatt gccataccca caaattttac tattagtgtt accacagaaa ttctaccagt
     5581 gtctatgacc aagacatcag tagattgtac aatgtacatt tgtggtgatt caactgaatg
     5641 cagcaatctt ttgttgcaat atggcagttt ttgtacacaa ttaaaccgtg ctttaactgg
     5701 aatagctgtt gaacaagaca aaaacaccca agaagttttt gcacaagtca aacaaattta
     5761 caaaacacca ccaattaaag attttggtgg ttttaatttt tcacaaatat taccagatcc
     5821 atcaaaacca agcaagaggt catttattga agatctactt ttcaacaaag tgacacttgc
     5881 agatgctggc ttcatcaaac aatatggtga ttgccttggt gatattgctg ctagagacct
     5941 catttgtgca caaaagttta acggccttac tgttttgcca cctttgctca cagatgaaat
     6001 gattgctcaa tacacttctg cactgttagc gggtacaatc acttctggtt ggacctttgg
     6061 tgcaggtgct gcattacaaa taccatttgc tatgcaaatg gcttataggt ttaatggtat
     6121 tggagttaca cagaatgttc tctatgagaa ccaaaaattg attgccaacc aatttaatag
     6181 tgctattggc aaaattcaag actcactttc ttccacagca agtgcacttg gaaaacttca
     6241 agatgtggtc aaccaaaatg cacaagcttt aaacacgctt gttaaacaac ttagctccaa
     6301 ttttggtgca atttcaagtg ttttaaatga tatcctttca cgtcttgaca aagttgaggc
     6361 tgaagtgcaa attgataggt tgatcacagg cagacttcaa agtttgcaga catatgtgac
     6421 tcaacaatta attagagctg cagaaatcag agcttctgct aatcttgctg ctactaaaat
     6481 gtcagagtgt gtacttggac aatcaaaaag agttgatttt tgtggaaagg gctatcatct
     6541 tatgtccttc cctcagtcag cacctcatgg tgtagtcttc ttgcatgtga cttatgtccc
     6601 tgcacaagaa aagaacttca caactgctcc tgccatttgt catgatggaa aagcacactt
     6661 tcctcgtgaa ggtgtctttg tttcaaatgg cacacactgg tttgtaacac aaaggaattt
     6721 ttatgaacca caaatcatta ctacagacaa cacatttgtg tctggtaact gtgatgttgt
     6781 aataggaatt gtcaacaaca cagtttatga tcctttgcaa cctgaattag actcattcaa
     6841 ggaggagtta gataaatatt ttaagaatca tacatcacca gatgttgatt taggtgacat
     6901 ctctggcatt aatgcttcag ttgtaaacat tcaaaaagaa attgaccgcc tcaatgaggt
     6961 tgccaagaat ttaaatgaat ctctcatcga tctccaagaa cttggaaagt atgagcagta
     7021 tataaaatgg ccatggtaca tttggctagg ttttatagct ggcttgattg ccatagtaat
     7081 ggtgacaatt atgctttgct gtatgaccag ttgctgtagt tgtctcaagg gctgttgttc
     7141 ttgtggatcc tgctgcaaat ttgatgaaga cgactctgag ccagtgctca aaggagtcaa
     7201 attacattac acagaaaacc tttactttca gtcgggacgg cgaatcccgg ggctgataaa
     7261 tccctggaaa aggagatgga agaagaactt tattgcagtg tctgcggcta accggttcaa
     7321 aaagatcagt tcaagcggcg ccctagatta cgacattcct acgaccgcct ccgagaatct
     7381 ctatttccag ggtgaatttg gcttggcaca acatgacgag gccgtcgata acaagtttaa
     7441 taaagaacag cagaacgctt tctatgagat ccttcactta ccaaatctga acgaagagca
     7501 acgcaacgct ttcattcaga gcctgaaaga cgatccctcg cagtctgcaa atctcctggc
     7561 cgaagccaag aagctgaatg acgcacaggc tccaaaagtt gataacaagt ttaacaagga
     7621 gcagcagaat gctttctacg agatattgca ccttcctaat ctgaatgagg aacaaagaaa
     7681 tgcatttatc cagtccctaa aagacgatcc atcacagagt gccaacctgc ttgcggaagc
     7741 caaaaagctc aatgatgccc aggccccgaa agtagacgcg aaccatcaat aagctagctt
     7801 gactgactga gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt
     7861 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     7921 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     7981 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     8041 ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct
     8101 aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg
     8161 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     8221 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     8281 ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc
     8341 tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc
     8401 tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta
     8461 cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg
     8521 atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc
     8581 aaa
//
