LOCUS       sequence-459016-        8607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of SARS-CoV-2 spike protein with glycan deletion at 
            N1098, N1173 by implementing Asn-to-Gln mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8607)
  AUTHORS   Yang Q, Kelkar A, Manicassamy B, Neelamegham S
  TITLE     Conserved role of spike S2 domain N-glycosylation across 
            betacoronaviruses.
  JOURNAL   Npj Viruses. 2025 Jan 25;3(1):4. doi: 10.1038/s44298-024-00085-7.
  PUBMED    40295734
REFERENCE   2  (bases 1 to 8607)
  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: "Npj 
            Viruses."; date: "2025-01-25"; volume: "3(1):4. doi"; pages: " 
            10.1038/s44298-024-00085-7"
COMMENT     Sequence Label: SARS-CoV-2 spike G14 mutant
FEATURES             Location/Qualifiers
     source          1..8607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          668..1688
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1608..1630
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1696..1715
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             5577..5600
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5628..5647)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5693..5748
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5747..5766)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6109..6125)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6109..6125)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6122..6144)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6133..6149
                     /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(6157..6163,6164..6181,6182..6187))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6202..6223
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6282..6477
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6328..6463
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6483..6617
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6520..6539)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6574..6593
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6685..6702)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6856..7444)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6936..6955)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7615..8406,8407..8475))
                     /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     8238..8257
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8476..8580)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg gggggggggg ccgggggggg gggggggggg gggggggggg
      541 gggggggggg ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg
      601 aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg
      661 gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg
      721 ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc
      781 ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct
      841 gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg
      901 tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg
      961 tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc
     1021 ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg
     1081 gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc
     1141 cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac
     1201 ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg
     1261 cggggcgggg ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag
     1321 cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga
     1381 gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg
     1441 ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg
     1501 cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg
     1561 ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg
     1621 gcgtgtgacc ggcggctcta gctagagcct ctgctaacca tgttcatgcc ttcttctttt
     1681 tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat
     1741 tcgccgccac catgtttgtg ttcctggtcc tgctcccact ggtttccagc caatgtgtta
     1801 acctcacaac ccgcacccag ctgcctcccg cctataccaa cagttttact cggggagtgt
     1861 attaccctga caaagttttc aggagctcag ttctgcactc aacccaggat ctctttctgc
     1921 cttttttttc caacgtgact tggttccatg caattcacgt ttccgggacc aacggcacca
     1981 aaaggttcga taaccccgtg ctgcccttta acgacggcgt ctatttcgca agcactgaga
     2041 aaagcaatat catcaggggc tggatttttg ggacaacctt ggatagcaag acccagtcac
     2101 tgctgatcgt taataatgca accaatgtgg tgatcaaagt gtgcgaattt cagttttgca
     2161 atgacccctt tctgggcgtc tactatcata aaaacaataa atcttggatg gagtctgagt
     2221 ttagggtgta ttctagcgcg aataattgta cctttgaata tgtatcacaa ccatttctca
     2281 tggatctcga aggcaaacaa ggcaatttca aaaacttgag agagttcgtc ttcaaaaaca
     2341 tcgatggata ttttaaaata tacagcaaac ataccccaat aaacctggtc cgcgacctcc
     2401 ctcagggttt cagcgccctc gagcctctcg tcgatcttcc tatcgggatc aacatcacaa
     2461 gatttcaaac cttgctggct cttcatcgat cctacctgac ccctggcgat tcctcttccg
     2521 ggtggaccgc cggcgccgct gcctattatg tggggtatct gcaacctcgc acgtttctcc
     2581 tcaagtataa tgagaatgga actatcaccg atgcggtaga ctgtgcgttg gacccactct
     2641 ctgagaccaa gtgcactttg aagtccttta ccgttgaaaa ggggatctat cagacatcca
     2701 attttagagt ccaacccacg gaatccatcg ttcggttccc caatatcaca aatctctgcc
     2761 ctttcggaga ggtattcaac gcaactcgct tcgcttctgt gtatgcgtgg aacagaaaac
     2821 gcatctccaa ctgcgtcgct gattactctg ttctgtataa ctccgcctcc tttagcacct
     2881 tcaagtgtta cggcgtcagc cccacaaagc tcaacgactt gtgtttcact aacgtctacg
     2941 ccgactcctt cgtgatccgg ggggacgaag ttcggcagat tgcgcctgga cagactggta
     3001 aaatcgctga ctataactat aagctcccag atgactttac cggctgtgtg attgcttgga
     3061 actctaataa tctcgacagc aaagttggag gcaactataa ttatctctac cggttgtttc
     3121 ggaagagcaa tctcaagcca ttcgagagag acataagcac ggagatctac caagccggct
     3181 ccactccatg caatggggtt gaaggcttta attgttattt tcccctccaa tcctatggtt
     3241 ttcaaccgac taatggagtc gggtatcaac cctacagggt tgtcgtcttg agcttcgagc
     3301 tcttgcacgc tcctgctacg gtgtgtggtc cgaaaaaatc aacaaacctc gtcaaaaaca
     3361 aatgtgtgaa ttttaacttt aacggactga ctggcaccgg ggtgctgact gagagcaata
     3421 agaagtttct tccatttcaa caattcggtc gggatatcgc ggacaccact gatgctgtgc
     3481 gcgatccgca aaccttggag atcttggaca taactccgtg cagcttcgga ggcgtctctg
     3541 ttatcacccc tggaacgaat acatctaacc aagttgcggt cctttatcag ggcgttaact
     3601 gcacagaagt acccgtcgcg atacatgcgg atcagctgac ccctacttgg cgggtctact
     3661 caaccggctc taatgtgttt caaactcggg cgggatgttt gatcggagct gagcacgtga
     3721 ataactctta tgaatgcgat ataccaatcg gcgcagggat ttgcgcatct taccaaaccc
     3781 aaaccaactc tcctcggcgg gctcgctccg tcgcatccca atccattatc gcctatacca
     3841 tgagtctcgg agctgagaac tctgtcgctt acagcaacaa ttctatagcc atcccaacca
     3901 acttcacgat tagcgttaca acagagatac tccccgtcag catgacgaaa acttccgttg
     3961 attgtacgat gtatatctgt ggggattcta cagagtgtag caatctgctg ttgcagtatg
     4021 gctccttctg tacccaattg aatcgggcac tcaccggcat cgctgtggaa caagacaaaa
     4081 atacgcaaga agttttcgca caggtgaagc aaatctacaa gacgcctcca atcaaagact
     4141 ttggcggatt caatttttct cagattctgc ctgatccctc aaagcccagc aaaagaagct
     4201 tcatcgagga cctgctcttc aacaaggtca ctctcgcaga tgccgggttc atcaaacaat
     4261 atggagattg cttgggcgac attgctgcgc gggatcttat ttgtgctcaa aagttcaatg
     4321 ggctgaccgt cctgccgccc ctcttgaccg atgaaatgat tgctcaatac acctccgcgt
     4381 tgctcgcggg aactatcact tccggctgga ccttcggcgc cggcgctgcc ctccagatac
     4441 cgtttgccat gcaaatggcc tacagattca acggcatcgg agtcacacag aatgtcctgt
     4501 acgaaaatca aaaattgata gccaatcagt ttaattctgc aatcggcaaa atacaagact
     4561 ccctgtcctc tactgcctcc gcacttggga aattgcaaga cgtcgtgaac caaaacgcac
     4621 aggcccttaa cactctggtg aaacagttga gctccaactt cggcgctata agctccgtct
     4681 tgaacgatat actctctcgc ctggacaaag tcgaggctga agtccagatc gacagattga
     4741 taacaggcag gcttcaatcc ctgcaaacct acgtcacgca gcagctgatc agagccgcag
     4801 agatcagggc ttccgcaaat cttgctgcaa caaaaatgtc cgaatgcgtc ttgggccaat
     4861 caaagcgcgt cgatttctgt ggaaagggct accatctcat gagctttccc caaagcgccc
     4921 cgcacggcgt ggtttttctc cacgtcacct acgtccccgc tcaagagaag aactttacaa
     4981 ccgctcctgc aatttgccat gatgggaaag cgcactttcc tcgcgaaggc gtctttgtct
     5041 cccagggcac acattggttt gttactcaac ggaactttta tgaaccacaa atcattacca
     5101 cggacaacac gttcgtgtcc ggcaactgtg acgttgtcat cggaattgtt aacaacacgg
     5161 tttacgatcc tctgcaaccc gagcttgact cctttaagga agaactggac aagtacttca
     5221 aaaatcacac ctcaccggat gtggacttgg gcgatatctc tggcatacag gcgtcagtcg
     5281 tgaacattca aaaggaaatt gatcgactca acgaggtcgc aaagaatctt aatgaatccc
     5341 tgatagatct gcaggagttg gggaagtacg agcagtatat aaagtggcct tggtatatat
     5401 ggcttggctt tatcgccggc ctcattgcaa tcgtgatggt caccatcatg ctgtgctgca
     5461 tgacctcatg ttgctcctgt cttaaggggt gctgctcctg cggttcctgt tgcaaatttg
     5521 acgaggacga cagcgaaccg gtgcttaagg gtgtgaagct gcattacacc ggaggcgatt
     5581 acaaggatga cgatgacaag tgacggccgc ctgcaggtta attaagatct ttttccctct
     5641 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
     5701 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
     5761 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
     5821 tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg
     5881 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
     5941 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
     6001 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
     6061 ctcttatgga gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt
     6121 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6181 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6241 gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca
     6301 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     6361 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     6421 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     6481 ctaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6541 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6601 cttatcatgt ctggatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6661 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6721 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6781 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6841 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6901 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6961 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     7021 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     7081 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     7141 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     7201 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7261 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     7321 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7381 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7441 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7501 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7561 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7621 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7681 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7741 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7801 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7861 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7921 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7981 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     8041 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     8101 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     8161 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     8221 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8281 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8341 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8401 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8461 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8521 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     8581 cacatttccc cgaaaagtgc cacctgg
//
