LOCUS       sequence-340758-        8377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SARS-CoV-2 Spike protein P-1-like variant 
            (K417N).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8377)
  AUTHORS   Gobeil SM, Janowska K, McDowell S, Mansouri K, Parks R, Stalls V, 
            Kopp MF, Manne K, Li D, Wiehe K, Saunders KO, Edwards RJ, Korber B, 
            Haynes BF, Henderson R, Acharya P
  TITLE     Effect of natural mutations of SARS-CoV-2 on spike structure, 
            conformation, and antigenicity.
  JOURNAL   Science. 2021 Jun 24. pii: science.abi6226. doi: 
            10.1126/science.abi6226.
  PUBMED    34168071
REFERENCE   2  (bases 1 to 8377)
  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"; journalName: "Science. 
            2021 Jun 24. pii: science.abi6226. doi: 10.1126/science.abi6226."
COMMENT     Sequence Label: p-alpha-H-S-GSAS-P1-like
FEATURES             Location/Qualifiers
     source          1..8377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             5487..5510
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5514..5537
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             5544..5567
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             5607..5630
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(5732..5751)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5797..5852
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5851..5870)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6213..6229)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6213..6229)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6226..6248)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6237..6253
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6261..6267,6268..6285,6286..6291))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6306..6327
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6456..6473)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6627..7215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6707..6726)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7386..8177,8178..8246))
                     /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     8009..8028
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8247..8351)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggnsgcg cgccrggsgg ggsggggsgg ggsgrggggs
      541 ggggsggggs gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagcgcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt gttgtgctgt ctcatcattt tggcaaagaa ttgcggccgt
     1741 ctcaggccga gttcggtacc gccaccatgt tcgtgttcct ggtgctcctg cctctggtga
     1801 gcagccagtg cgtgaacttc accaaccgaa cccagctccc aagcgcctac accaacagct
     1861 ttacacgggg cgtgtactac cctgacaagg tgttcagatc tagcgtcctg cacagcactc
     1921 aggacctctt cctgccgttc ttcagcaacg tgacatggtt ccacgccatc cacgtgagcg
     1981 gcacaaacgg aaccaagcgg tttgataacc ccgtcctgcc attcaatgat ggagtttact
     2041 tcgccagtac cgagaagagt aacatcatcc ggggctggat cttcggcacc accctggata
     2101 gcaaaacaca gagcctcctg atcgtgaaca atgccacgaa cgtcgtgatc aaggtgtgcg
     2161 agttccagtt ttgcaattat cctttcctgg gtgtgtacta ccacaagaac aacaagagct
     2221 ggatggaaag cgagttcaga gtctacagca gcgccaacaa ctgcacattc gagtacgtct
     2281 ctcagccttt tctgatggac cttgagggga aacaaggcaa cttcaagaac ctgagcgaat
     2341 tcgtgttcaa gaacatcgac ggctacttca aaatctactc caagcacaca cccatcaacc
     2401 tggtccggga cctccctcag ggcttcagcg ccctggaacc cctggtcgac ctgcccatag
     2461 gcatcaacat aacgcggttc caaaccctgc tggccctgca tagatcctac ctgactcctg
     2521 gcgacagcag cagcggatgg accgccggag ctgcagccta ctatgtgggc tacctgcaac
     2581 ctagaacctt cctgctgaag tacaacgaga acggcacaat cacagacgcc gtcgactgcg
     2641 ccctggaccc tctctctgag acaaagtgca ccctgaagtc cttcaccgtg gaaaagggca
     2701 tctaccagac cagcaacttc cgggtgcagc ctacagagag catcgtgcga tttccaaaca
     2761 ttaccaacct ctgccccttc ggcgaggtgt ttaacgccac aagatttgcc tccgtttacg
     2821 cctggaatag aaagagaatc agcaattgtg tggccgacta ctccgtgctg tataacagcg
     2881 cctctttcag caccttcaag tgctacggcg tttccccaac aaagctgaat gacctgtgct
     2941 tcaccaacgt gtacgccgac tccttcgtaa ttagaggcga tgaggtgcgg cagatcgcac
     3001 caggccagac cggtaacatc gctgactaca actataagct gcctgatgat tttacaggct
     3061 gcgtgatcgc ctggaactct aacaacctgg atagcaaggt gggcggcaac tacaactacc
     3121 tgtaccggct gtttcgcaag tctaacctga aacctttcga gagagacatc tccacagaga
     3181 tctaccaggc cggttctaca ccttgtaacg gggtgaaagg cttcaactgt tacttccctc
     3241 tgcaaagcta cggcttccag cctacctacg gagtcggcta ccagccatac cgggtggtcg
     3301 tgctgtcctt cgagttactc cacgcccccg ccaccgtctg cggtcctaag aagtccacca
     3361 atctggttaa gaacaaatgc gtgaacttca acttcaacgg cctgaccggg accggcgtgc
     3421 tgaccgaaag caacaaaaag ttcctcccct tccagcagtt cggccgtgat atcgctgaca
     3481 ccacagatgc cgtcagagat ccacagaccc tggaaatcct ggatattaca ccctgctcct
     3541 tcggaggagt ttctgtgatc acccccggga ccaataccag caaccaggtg gctgtgctgt
     3601 accaaggtgt taactgcacc gaggttcctg tggccatcca cgccgatcag ctgacaccta
     3661 cttggagagt gtactccact ggctccaatg tgttccagac cagggccgga tgtctgatcg
     3721 gcgccgagta cgtgaataac agttacgagt gcgacatccc tatcggcgcc ggcatctgtg
     3781 ccagctacca gacccagaca aacagccctg ggtctgcttc ctctgtagct agccagagca
     3841 tcatcgccta caccatgagc ctgggcgcag agaacagcgt ggcctattcc aacaactcta
     3901 tcgccattcc caccaacttt acaattagcg tcacaacaga gatcctgccc gtgagcatga
     3961 ccaagaccag cgtggactgt acaatgtaca tctgtggcga cagcactgaa tgcagcaacc
     4021 tgctgctgca atacggctcc ttttgcaccc aactgaaccg ggcgctgacc ggaatcgccg
     4081 tggaacagga caaaaatacc caggaggtgt tcgcccaagt gaagcagatc tacaagaccc
     4141 cacctatcaa ggacttcggc ggctttaact ttagccagat tctccctgat ccttctaagc
     4201 ctagcaagcg gagctttatc gaggatctgc tgttcaacaa ggtcaccctg gccgatgccg
     4261 gctttatcaa acagtatggc gattgcctgg gcgacatagc cgccagagat ctgatctgcg
     4321 cccagaaatt caacggcctg acagttctcc cacctctgct gaccgacgag atgatcgctc
     4381 agtacacctc tgccctgctg gctggcacca tcacatctgg gtggacattt ggcgccggcg
     4441 ccgccctgca gatccccttt gccatgcaga tggcctatag attcaacgga atcggcgtga
     4501 cccagaacgt gctgtatgaa aaccagaagc tgatcgctaa ccagttcaat tctgccatcg
     4561 gcaagatcca ggactccctc tcctctaccg ccagcgccct gggcaaactg caggacgtgg
     4621 tgaatcagaa cgcccaagcc ctgaacaccc tggtgaagca gctcagcagc aattttggcg
     4681 ccatcagctc tgtgctgaac gatatcctgt ctagactgga caaggtggaa gccgaagtcc
     4741 agatcgatag actgatcaca ggcagactgc agtccctgca aacctacgtg acccaacagc
     4801 tgatcagggc cgctgaaata agagccagcg ccaatctcgc cgctatcaag atgtccgagt
     4861 gtgtgctggg acagtctaaa cgcgttgact tctgcggcaa aggctatcac ctgatgagct
     4921 tcccccagag cgcgccgcac ggcgtggtgt tcctgcatgt gacatacgtg cctgcccaag
     4981 agaagaattt cacaaccgcc cctgccatct gccacgacgg caaggcccac ttcccaagag
     5041 agggcgtttt cgtttccaat ggcacacact ggttcgtgac acaaagaaac ttctacgaac
     5101 cccagattat caccaccgac aacaccttcg tgagtggcaa ttgtgacgtg gtcatcggaa
     5161 tcgtgaacaa cacagtgtac gaccctctgc aacctgagct ggactctttt aaggaagagc
     5221 tggacaagta ctttaaaaac cacaccagcc ccgatgtgga cctgggcgac atcagtggca
     5281 ttaacgccag cgtggtgaac atccaaaagg aaatcgacag actgaacgag gtggccaaga
     5341 acctgaacga gtccctgatc gacctgcagg agctcggcaa atacgagcag ggatccggat
     5401 acatccccga ggcccccaga gatggccagg cctacgtgcg gaaggacggc gagtgggtac
     5461 tgctgagcac attcctgggc agatccctgg aggtgctgtt ccagggccca ggccatcacc
     5521 accatcacca ccatcatagc gcctggtccc acccccagtt cgagaagggc ggcggtagtg
     5581 gagggggcgg atctggcggc tcagcttgga gccaccccca gttcgaaaag tgataatgac
     5641 tcgagcgata attcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg
     5701 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg
     5761 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     5821 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     5881 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     5941 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     6001 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     6061 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     6121 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc
     6181 tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6241 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6301 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6361 ggaaacctgt cgtgccagcg gatcgatccg ctgcattaat gaatcggcca acgcgcgggg
     6421 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6481 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6541 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6601 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6661 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6721 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6781 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6841 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6901 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6961 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7021 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     7081 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7141 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7201 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7261 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7321 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7381 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7441 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7501 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7561 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7621 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7681 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7741 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7801 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7861 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7921 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7981 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8041 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8101 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8161 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8221 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8281 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8341 ggggttccgc gcacatttcc ccgaaaagtg ccacctg
//
