LOCUS       sequence-300222-        9677 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 spike 
            prefusion-stabilized.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9677)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 9677)
  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-m4134900
FEATURES             Location/Qualifiers
     source          1..9677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    3639..3760
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3650..3669)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3704..3723
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3875..3921
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3951..4087
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4132..4203
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4429..5017)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4509..4528)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5517..6726
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6563..6582
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             6730..7329
                     /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(6730..6749)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7226..7246
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7356..7404
                     /note="synthetic polyadenylation signal"
     CDS             join(7670..7738,7739..8530)
                     /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(7888..7907)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      8543..8679
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8831..8902
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8907..8953)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    8971..9019
                     /note="synthetic polyadenylation signal"
     enhancer        9100..9403
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9405..9607
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9558..9578
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(9676..9677,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggtt aatcttacaa ccagaactca attaccccct gcatacacta attctttcac
       61 acgtggtgtt tattaccctg acaaagtttt cagatcctca gttttacatt caactcagga
      121 cttgttctta cctttctttt ccaatgttac ttggttccat gctatacatg tctctgggac
      181 caatggtact aagaggtttg ataaccctgt cctaccattt aatgatggtg tttattttgc
      241 ttccactgag aagtctaaca taataagagg ctggattttt ggtactactt tagattcgaa
      301 gacccagtcc ctacttattg ttaataacgc tactaatgtt gttattaaag tctgtgaatt
      361 tcaattttgt aatgatccat ttttgggtgt ttattaccac aaaaacaaca aaagttggat
      421 ggaaagtgag ttcagagttt attctagtgc gaataattgc acttttgaat atgtctctca
      481 gccttttctt atggaccttg aaggaaaaca gggtaatttc aaaaatctta gggaatttgt
      541 gtttaagaat attgatggtt attttaaaat atattctaag cacacgccta ttaatttagt
      601 gcgtgatctc cctcagggtt tttcggcttt agaaccattg gtagatttgc caataggtat
      661 taacatcact aggtttcaaa ctttacttgc tttacataga agttatttga ctcctggtga
      721 ttcttcttca ggttggacag ctggtgctgc agcttattat gtgggttatc ttcaacctag
      781 gacttttcta ttaaaatata atgaaaatgg aaccattaca gatgctgtag actgtgcact
      841 tgaccctctc tcagaaacaa agtgtacgtt gaaatccttc actgtagaaa aaggaatcta
      901 tcaaacttct aactttagag tccaaccaac agaatctatt gttagatttc ctaatattac
      961 aaacttgtgc ccttttggtg aagtttttaa cgccaccaga tttgcatctg tttatgcttg
     1021 gaacaggaag agaatcagca actgtgttgc tgattattct gtcctatata attccgcatc
     1081 attttccact tttaagtgtt atggagtgtc tcctactaaa ttaaatgatc tctgctttac
     1141 taatgtctat gcagattcat ttgtaattag aggtgatgaa gtcagacaaa tcgctccagg
     1201 gcaaactgga aagattgctg attataatta taaattacca gatgatttta caggctgcgt
     1261 tatagcttgg aattctaaca atcttgattc taaggttggt ggtaattata attacctgta
     1321 tagattgttt aggaagtcta atctcaaacc ttttgagaga gatatttcaa ctgaaatcta
     1381 tcaggccggt agcacacctt gtaatggtgt tgaaggtttt aattgttact ttcctttaca
     1441 atcatatggt ttccaaccca ctaatggtgt tggttaccaa ccatacagag tagtagtact
     1501 ttcttttgaa cttctacatg caccagcaac tgtttgtgga cctaaaaagt ctactaattt
     1561 ggttaaaaac aaatgtgtca atttcaactt caatggttta acaggcacag gtgttcttac
     1621 tgagtctaac aaaaagtttc tgcctttcca acaatttggc agagacattg ctgacactac
     1681 tgatgctgtc cgtgatccac agacacttga gattcttgac attacaccat gttcttttgg
     1741 tggtgtcagt gttataacac caggaacaaa tacttctaac caggttgctg ttctttatca
     1801 ggatgttaac tgcacagaag tccctgttgc tattcatgca gatcaactta ctcctacttg
     1861 gcgtgtttat tctacaggtt ctaatgtttt tcaaacacgt gcaggctgtt taataggggc
     1921 tgaacatgtc aacaactcat atgagtgtga catacccatt ggtgcaggta tatgcgctag
     1981 ttatcagact cagactaatt ctcctgcacg tgcaggtagt gtagctagtc aatccatcat
     2041 tgcctacact atgtcacttg gtgcagaaaa ttcagttgct tactctaata actctattgc
     2101 catacccaca aattttacta ttagtgttac cacagaaatt ctaccagtgt ctatgaccaa
     2161 gacatcagta gattgtacaa tgtacatttg tggtgattca actgaatgca gcaatctttt
     2221 gttgcaatat ggcagttttt gtacacaatt aaaccgtgct ttaactggaa tagctgttga
     2281 acaagacaaa aacacccaag aagtttttgc acaagtcaaa caaatttaca aaacaccacc
     2341 aattaaagat tttggtggtt ttaatttttc acaaatatta ccagatccat caaaaccaag
     2401 caagaggtca tttattgaag atctactttt caacaaagtg acacttgcag atgctggctt
     2461 catcaaacaa tatggtgatt gccttggtga tattgctgct agagacctca tttgtgcaca
     2521 aaagtttaac ggccttactg ttttgccacc tttgctcaca gatgaaatga ttgctcaata
     2581 cacttctgca ctgttagcgg gtacaatcac ttctggttgg acctttggtg caggtgctgc
     2641 attacaaata ccatttgcta tgcaaatggc ttataggttt aatggtattg gagttacaca
     2701 gaatgttctc tatgagaacc aaaaattgat tgccaaccaa tttaatagtg ctattggcaa
     2761 aattcaagac tcactttctt ccacagcaag tgcacttgga aaacttcaag atgtggtcaa
     2821 ccaaaatgca caagctttaa acacgcttgt taaacaactt agctccaatt ttggtgcaat
     2881 ttcaagtgtt ttaaatgata tcctttcacg tcttgaccca ccagaggctg aagtgcaaat
     2941 tgataggttg atcacaggca gacttcaaag tttgcagaca tatgtgactc aacaattaat
     3001 tagagctgca gaaatcagag cttctgctaa tcttgctgct actaaaatgt cagagtgtgt
     3061 acttggacaa tcaaaaagag ttgatttttg tggaaagggc tatcatctta tgtccttccc
     3121 tcagtcagca cctcatggtg tagtcttctt gcatgtgact tatgtccctg cacaagaaaa
     3181 gaacttcaca actgctcctg ccatttgtca tgatggaaaa gcacactttc ctcgtgaagg
     3241 tgtctttgtt tcaaatggca cacactggtt tgtaacacaa aggaattttt atgaaccaca
     3301 aatcattact acagacaaca catttgtgtc tggtaactgt gatgttgtaa taggaattgt
     3361 caacaacaca gtttatgatc ctttgcaacc tgaattagac tcattcaagg aggagttaga
     3421 taaatatttt aagaatcata catcaccaga tgttgattta ggtgacatct ctggcattaa
     3481 tgcttcagtt gtaaacattc aaaaagaaat tgaccgcctc aatgaggttg ccaagaattt
     3541 aaatgaatct ctcatcgatc tccaagaact tggaaagtat gagcagtaag ctagcttgac
     3601 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
     3661 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3721 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3781 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     3841 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
     3901 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
     3961 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     4021 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     4081 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
     4141 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
     4201 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
     4261 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
     4321 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
     4381 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4441 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4501 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4561 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4621 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4681 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4741 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4801 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4861 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4921 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4981 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5041 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5101 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5161 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5221 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     5281 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     5341 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     5401 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     5461 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc
     5521 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac
     5581 gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
     5641 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
     5701 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     5761 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     5821 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     5881 ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg
     5941 gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg
     6001 gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca
     6061 aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg
     6121 ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta
     6181 atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg
     6241 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
     6301 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
     6361 tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg tgtgcggtag
     6421 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
     6481 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
     6541 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
     6601 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
     6661 aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg
     6721 ttagacaaca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     6781 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     6841 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     6901 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     6961 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     7021 ttgagcggtt cccggctggc cgcgcagcaa cagatggagg gcctcctggc gccgcaccgg
     7081 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     7141 ggtctgggca gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc cggggtgccc
     7201 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     7261 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     7321 ggtgcctagg ctagtatgta agcctagtct tagataataa aatcgctatc catcgaagat
     7381 ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat
     7441 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     7501 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     7561 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     7621 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca
     7681 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     7741 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta
     7801 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt
     7861 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     7921 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc
     7981 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     8041 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa
     8101 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     8161 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     8221 ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca
     8281 gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     8341 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     8401 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     8461 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     8521 gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     8581 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     8641 cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc gtttaaactc
     8701 gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat
     8761 ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg
     8821 cggctcccga catcttggac cattagctcc acaggtatct tcttccctct agtggtcata
     8881 acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca
     8941 aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc
     9001 gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat
     9061 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     9121 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     9181 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     9241 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     9301 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     9361 tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca
     9421 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     9481 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     9541 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     9601 cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca
     9661 cacccgccag cggccgc
//
