LOCUS       sequence-304902-       10013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 surface glycoprotein 
            (Spike protein).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10013)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 10013)
  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-m4134122
FEATURES             Location/Qualifiers
     source          1..10013
                     /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             3845..3922
                     /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"
     polyA_signal    3975..4096
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3986..4005)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4040..4059
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4211..4257
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4287..4423
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4468..4539
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4765..5353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4845..4864)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5853..7062
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6899..6918
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             7066..7665
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7066..7085)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7562..7582
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7692..7740
                     /note="synthetic polyadenylation signal"
     CDS             join(8006..8074,8075..8866)
                     /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(8224..8243)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      8879..9015
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    9167..9238
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(9243..9289)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    9307..9355
                     /note="synthetic polyadenylation signal"
     enhancer        9436..9739
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9741..9943
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9894..9914
                     /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 gtcgaagagg cgatggaaaa agaattttat cgcggtgagt gcagccaaca gattcaaaaa
     3901 gatatcctcg tctggtgccc tctaagctag cttgactgac tgagatacag cgtaccttca
     3961 gctcacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     4021 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     4081 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     4141 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct
     4201 atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc
     4261 ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc
     4321 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     4381 cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg
     4441 caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta
     4501 ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc
     4561 tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt
     4621 cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc
     4681 tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4741 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4801 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4861 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4921 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4981 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5041 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5101 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5161 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5221 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5281 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5341 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5401 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5461 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5521 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5581 catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt
     5641 tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc
     5701 cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata
     5761 tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga
     5821 gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc
     5881 cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt
     5941 cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta
     6001 gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg
     6061 gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg
     6121 gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca
     6181 cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat
     6241 cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc
     6301 tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa
     6361 ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt
     6421 gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt
     6481 gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc ttattcgggt
     6541 gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact
     6601 cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt
     6661 acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat
     6721 gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag
     6781 ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag
     6841 ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg
     6901 aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag
     6961 gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gacttgtaaa
     7021 ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatgac cgagtacaag
     7081 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     7141 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     7201 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     7261 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     7321 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     7381 cagcaacaga tggagggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     7441 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     7501 cccggagtgg aggccgccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     7561 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     7621 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctaggctag tatgtaagcc
     7681 tagtcttaga taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg
     7741 tgtaacgcta ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat
     7801 ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat
     7861 cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata
     7921 ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga
     7981 gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc
     8041 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     8101 agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg
     8161 taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt
     8221 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     8281 catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac
     8341 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     8401 ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa
     8461 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     8521 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt
     8581 aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga
     8641 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     8701 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     8761 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     8821 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc
     8881 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca
     8941 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca
     9001 gaaatttatc cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg
     9061 tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg
     9121 tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt
     9181 agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa
     9241 ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc
     9301 gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta
     9361 actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc
     9421 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     9481 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     9541 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     9601 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     9661 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     9721 agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     9781 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     9841 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     9901 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
     9961 gatcagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc cgc
//
