LOCUS       sequence-459015-        8602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of SARS-CoV-2 spike protein with glycan deletion at 
            N1098, N1194 by implementing Asn-to-Gln mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8602)
  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 8602)
  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 G15 mutant
FEATURES             Location/Qualifiers
     source          1..8602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..663
                     /label=chicken beta-actin promoter
     intron          664..1684
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1604..1626
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1692..1711
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             5573..5596
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5624..5643)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5689..5744
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5743..5762)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6105..6121)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6105..6121)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6118..6140)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6129..6145
                     /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(6153..6159,6160..6177,6178..6183))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6198..6219
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6278..6473
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6324..6459
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6479..6613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6516..6535)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6570..6589
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6681..6698)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6852..7440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6932..6951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7611..8402,8403..8471))
                     /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     8234..8253
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8472..8576)
                     /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 gggggggggc gcgcccgggg gggggggggg gggggggggg
      541 gggggggggg ggaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
      601 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
      661 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
      721 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
      781 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
      841 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag
      961 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc
     1021 gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt
     1081 gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg
     1141 cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg
     1201 cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg
     1261 gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc
     1321 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
     1381 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
     1441 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
     1501 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt
     1561 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
     1621 gtgaccggcg gctctagcta gagcctctgc taaccatgtt catgccttct tctttttcct
     1681 acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcgc
     1741 cgccaccatg tttgtgttcc tggtcctgct cccactggtt tccagccaat gtgttaacct
     1801 cacaacccgc acccagctgc ctcccgccta taccaacagt tttactcggg gagtgtatta
     1861 ccctgacaaa gttttcagga gctcagttct gcactcaacc caggatctct ttctgccttt
     1921 tttttccaac gtgacttggt tccatgcaat tcacgtttcc gggaccaacg gcaccaaaag
     1981 gttcgataac cccgtgctgc cctttaacga cggcgtctat ttcgcaagca ctgagaaaag
     2041 caatatcatc aggggctgga tttttgggac aaccttggat agcaagaccc agtcactgct
     2101 gatcgttaat aatgcaacca atgtggtgat caaagtgtgc gaatttcagt tttgcaatga
     2161 cccctttctg ggcgtctact atcataaaaa caataaatct tggatggagt ctgagtttag
     2221 ggtgtattct agcgcgaata attgtacctt tgaatatgta tcacaaccat ttctcatgga
     2281 tctcgaaggc aaacaaggca atttcaaaaa cttgagagag ttcgtcttca aaaacatcga
     2341 tggatatttt aaaatataca gcaaacatac cccaataaac ctggtccgcg acctccctca
     2401 gggtttcagc gccctcgagc ctctcgtcga tcttcctatc gggatcaaca tcacaagatt
     2461 tcaaaccttg ctggctcttc atcgatccta cctgacccct ggcgattcct cttccgggtg
     2521 gaccgccggc gccgctgcct attatgtggg gtatctgcaa cctcgcacgt ttctcctcaa
     2581 gtataatgag aatggaacta tcaccgatgc ggtagactgt gcgttggacc cactctctga
     2641 gaccaagtgc actttgaagt cctttaccgt tgaaaagggg atctatcaga catccaattt
     2701 tagagtccaa cccacggaat ccatcgttcg gttccccaat atcacaaatc tctgcccttt
     2761 cggagaggta ttcaacgcaa ctcgcttcgc ttctgtgtat gcgtggaaca gaaaacgcat
     2821 ctccaactgc gtcgctgatt actctgttct gtataactcc gcctccttta gcaccttcaa
     2881 gtgttacggc gtcagcccca caaagctcaa cgacttgtgt ttcactaacg tctacgccga
     2941 ctccttcgtg atccgggggg acgaagttcg gcagattgcg cctggacaga ctggtaaaat
     3001 cgctgactat aactataagc tcccagatga ctttaccggc tgtgtgattg cttggaactc
     3061 taataatctc gacagcaaag ttggaggcaa ctataattat ctctaccggt tgtttcggaa
     3121 gagcaatctc aagccattcg agagagacat aagcacggag atctaccaag ccggctccac
     3181 tccatgcaat ggggttgaag gctttaattg ttattttccc ctccaatcct atggttttca
     3241 accgactaat ggagtcgggt atcaacccta cagggttgtc gtcttgagct tcgagctctt
     3301 gcacgctcct gctacggtgt gtggtccgaa aaaatcaaca aacctcgtca aaaacaaatg
     3361 tgtgaatttt aactttaacg gactgactgg caccggggtg ctgactgaga gcaataagaa
     3421 gtttcttcca tttcaacaat tcggtcggga tatcgcggac accactgatg ctgtgcgcga
     3481 tccgcaaacc ttggagatct tggacataac tccgtgcagc ttcggaggcg tctctgttat
     3541 cacccctgga acgaatacat ctaaccaagt tgcggtcctt tatcagggcg ttaactgcac
     3601 agaagtaccc gtcgcgatac atgcggatca gctgacccct acttggcggg tctactcaac
     3661 cggctctaat gtgtttcaaa ctcgggcggg atgtttgatc ggagctgagc acgtgaataa
     3721 ctcttatgaa tgcgatatac caatcggcgc agggatttgc gcatcttacc aaacccaaac
     3781 caactctcct cggcgggctc gctccgtcgc atcccaatcc attatcgcct ataccatgag
     3841 tctcggagct gagaactctg tcgcttacag caacaattct atagccatcc caaccaactt
     3901 cacgattagc gttacaacag agatactccc cgtcagcatg acgaaaactt ccgttgattg
     3961 tacgatgtat atctgtgggg attctacaga gtgtagcaat ctgctgttgc agtatggctc
     4021 cttctgtacc caattgaatc gggcactcac cggcatcgct gtggaacaag acaaaaatac
     4081 gcaagaagtt ttcgcacagg tgaagcaaat ctacaagacg cctccaatca aagactttgg
     4141 cggattcaat ttttctcaga ttctgcctga tccctcaaag cccagcaaaa gaagcttcat
     4201 cgaggacctg ctcttcaaca aggtcactct cgcagatgcc gggttcatca aacaatatgg
     4261 agattgcttg ggcgacattg ctgcgcggga tcttatttgt gctcaaaagt tcaatgggct
     4321 gaccgtcctg ccgcccctct tgaccgatga aatgattgct caatacacct ccgcgttgct
     4381 cgcgggaact atcacttccg gctggacctt cggcgccggc gctgccctcc agataccgtt
     4441 tgccatgcaa atggcctaca gattcaacgg catcggagtc acacagaatg tcctgtacga
     4501 aaatcaaaaa ttgatagcca atcagtttaa ttctgcaatc ggcaaaatac aagactccct
     4561 gtcctctact gcctccgcac ttgggaaatt gcaagacgtc gtgaaccaaa acgcacaggc
     4621 ccttaacact ctggtgaaac agttgagctc caacttcggc gctataagct ccgtcttgaa
     4681 cgatatactc tctcgcctgg acaaagtcga ggctgaagtc cagatcgaca gattgataac
     4741 aggcaggctt caatccctgc aaacctacgt cacgcagcag ctgatcagag ccgcagagat
     4801 cagggcttcc gcaaatcttg ctgcaacaaa aatgtccgaa tgcgtcttgg gccaatcaaa
     4861 gcgcgtcgat ttctgtggaa agggctacca tctcatgagc tttccccaaa gcgccccgca
     4921 cggcgtggtt tttctccacg tcacctacgt ccccgctcaa gagaagaact ttacaaccgc
     4981 tcctgcaatt tgccatgatg ggaaagcgca ctttcctcgc gaaggcgtct ttgtctccca
     5041 gggcacacat tggtttgtta ctcaacggaa cttttatgaa ccacaaatca ttaccacgga
     5101 caacacgttc gtgtccggca actgtgacgt tgtcatcgga attgttaaca acacggttta
     5161 cgatcctctg caacccgagc ttgactcctt taaggaagaa ctggacaagt acttcaaaaa
     5221 tcacacctca ccggatgtgg acttgggcga tatctctggc ataaatgcgt cagtcgtgaa
     5281 cattcaaaag gaaattgatc gactcaacga ggtcgcaaag aatcttcagg aatccctgat
     5341 agatctgcag gagttgggga agtacgagca gtatataaag tggccttggt atatatggct
     5401 tggctttatc gccggcctca ttgcaatcgt gatggtcacc atcatgctgt gctgcatgac
     5461 ctcatgttgc tcctgtctta aggggtgctg ctcctgcggt tcctgttgca aatttgacga
     5521 ggacgacagc gaaccggtgc ttaagggtgt gaagctgcat tacaccggag gcgattacaa
     5581 ggatgacgat gacaagtgac ggccgcctgc aggttaatta agatcttttt ccctctgcca
     5641 aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat
     5701 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     5761 ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg
     5821 cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac
     5881 agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt
     5941 ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt
     6001 tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct
     6061 tatggagatc cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct
     6121 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6181 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6241 gctttccagt cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca
     6301 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     6361 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     6421 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa
     6481 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     6541 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     6601 tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6661 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6721 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6781 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6841 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6901 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6961 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7021 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     7081 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7141 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7201 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7261 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     7321 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7381 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7441 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7501 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7561 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7621 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7681 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7741 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7801 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7861 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7921 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7981 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8041 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8101 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8161 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8221 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8281 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8341 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8401 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8461 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8521 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8581 tttccccgaa aagtgccacc tg
//
