LOCUS       sequence-459014-        8602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of SARS-CoV-2 spike protein with glycan deletion at 
            N1098, N1134 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 G12 mutant
FEATURES             Location/Qualifiers
     source          1..8602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..662
                     /label=chicken beta-actin promoter
     intron          663..1683
                     /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"
     primer_bind     1691..1710
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             5572..5595
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5623..5642)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5688..5743
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5742..5761)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6104..6120)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6104..6120)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6117..6139)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6128..6144
                     /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(6152..6158,6159..6176,6177..6182))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6197..6218
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6277..6472
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6323..6458
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6478..6612
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6515..6534)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6569..6588
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6680..6697)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6851..7439)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6931..6950)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7610..8401,8402..8470))
                     /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     8233..8252
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8471..8575)
                     /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 cgatgggggg gggggggggg ggggggcgcg cggggggcgg gggggggcgg ggcggggggc
      541 ggggcggggc 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 ctctagctag agcctctgct aaccatgttc atgccttctt ctttttccta
     1681 cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattcgcc
     1741 gccaccatgt ttgtgttcct ggtcctgctc ccactggttt ccagccaatg tgttaacctc
     1801 acaacccgca cccagctgcc tcccgcctat accaacagtt ttactcgggg agtgtattac
     1861 cctgacaaag ttttcaggag ctcagttctg cactcaaccc aggatctctt tctgcctttt
     1921 ttttccaacg tgacttggtt ccatgcaatt cacgtttccg ggaccaacgg caccaaaagg
     1981 ttcgataacc ccgtgctgcc ctttaacgac ggcgtctatt tcgcaagcac tgagaaaagc
     2041 aatatcatca ggggctggat ttttgggaca accttggata gcaagaccca gtcactgctg
     2101 atcgttaata atgcaaccaa tgtggtgatc aaagtgtgcg aatttcagtt ttgcaatgac
     2161 ccctttctgg gcgtctacta tcataaaaac aataaatctt ggatggagtc tgagtttagg
     2221 gtgtattcta gcgcgaataa ttgtaccttt gaatatgtat cacaaccatt tctcatggat
     2281 ctcgaaggca aacaaggcaa tttcaaaaac ttgagagagt tcgtcttcaa aaacatcgat
     2341 ggatatttta aaatatacag caaacatacc ccaataaacc tggtccgcga cctccctcag
     2401 ggtttcagcg ccctcgagcc tctcgtcgat cttcctatcg ggatcaacat cacaagattt
     2461 caaaccttgc tggctcttca tcgatcctac ctgacccctg gcgattcctc ttccgggtgg
     2521 accgccggcg ccgctgccta ttatgtgggg tatctgcaac ctcgcacgtt tctcctcaag
     2581 tataatgaga atggaactat caccgatgcg gtagactgtg cgttggaccc actctctgag
     2641 accaagtgca ctttgaagtc ctttaccgtt gaaaagggga tctatcagac atccaatttt
     2701 agagtccaac ccacggaatc catcgttcgg ttccccaata tcacaaatct ctgccctttc
     2761 ggagaggtat tcaacgcaac tcgcttcgct tctgtgtatg cgtggaacag aaaacgcatc
     2821 tccaactgcg tcgctgatta ctctgttctg tataactccg cctcctttag caccttcaag
     2881 tgttacggcg tcagccccac aaagctcaac gacttgtgtt tcactaacgt ctacgccgac
     2941 tccttcgtga tccgggggga cgaagttcgg cagattgcgc ctggacagac tggtaaaatc
     3001 gctgactata actataagct cccagatgac tttaccggct gtgtgattgc ttggaactct
     3061 aataatctcg acagcaaagt tggaggcaac tataattatc tctaccggtt gtttcggaag
     3121 agcaatctca agccattcga gagagacata agcacggaga tctaccaagc cggctccact
     3181 ccatgcaatg gggttgaagg ctttaattgt tattttcccc tccaatccta tggttttcaa
     3241 ccgactaatg gagtcgggta tcaaccctac agggttgtcg tcttgagctt cgagctcttg
     3301 cacgctcctg ctacggtgtg tggtccgaaa aaatcaacaa acctcgtcaa aaacaaatgt
     3361 gtgaatttta actttaacgg actgactggc accggggtgc tgactgagag caataagaag
     3421 tttcttccat ttcaacaatt cggtcgggat atcgcggaca ccactgatgc tgtgcgcgat
     3481 ccgcaaacct tggagatctt ggacataact ccgtgcagct tcggaggcgt ctctgttatc
     3541 acccctggaa cgaatacatc taaccaagtt gcggtccttt atcagggcgt taactgcaca
     3601 gaagtacccg tcgcgataca tgcggatcag ctgaccccta cttggcgggt ctactcaacc
     3661 ggctctaatg tgtttcaaac tcgggcggga tgtttgatcg gagctgagca cgtgaataac
     3721 tcttatgaat gcgatatacc aatcggcgca gggatttgcg catcttacca aacccaaacc
     3781 aactctcctc ggcgggctcg ctccgtcgca tcccaatcca ttatcgccta taccatgagt
     3841 ctcggagctg agaactctgt cgcttacagc aacaattcta tagccatccc aaccaacttc
     3901 acgattagcg ttacaacaga gatactcccc gtcagcatga cgaaaacttc cgttgattgt
     3961 acgatgtata tctgtgggga ttctacagag tgtagcaatc tgctgttgca gtatggctcc
     4021 ttctgtaccc aattgaatcg ggcactcacc ggcatcgctg tggaacaaga caaaaatacg
     4081 caagaagttt tcgcacaggt gaagcaaatc tacaagacgc ctccaatcaa agactttggc
     4141 ggattcaatt tttctcagat tctgcctgat ccctcaaagc ccagcaaaag aagcttcatc
     4201 gaggacctgc tcttcaacaa ggtcactctc gcagatgccg ggttcatcaa acaatatgga
     4261 gattgcttgg gcgacattgc tgcgcgggat cttatttgtg ctcaaaagtt caatgggctg
     4321 accgtcctgc cgcccctctt gaccgatgaa atgattgctc aatacacctc cgcgttgctc
     4381 gcgggaacta tcacttccgg ctggaccttc ggcgccggcg ctgccctcca gataccgttt
     4441 gccatgcaaa tggcctacag attcaacggc atcggagtca cacagaatgt cctgtacgaa
     4501 aatcaaaaat tgatagccaa tcagtttaat tctgcaatcg gcaaaataca agactccctg
     4561 tcctctactg cctccgcact tgggaaattg caagacgtcg tgaaccaaaa cgcacaggcc
     4621 cttaacactc tggtgaaaca gttgagctcc aacttcggcg ctataagctc cgtcttgaac
     4681 gatatactct ctcgcctgga caaagtcgag gctgaagtcc agatcgacag attgataaca
     4741 ggcaggcttc aatccctgca aacctacgtc acgcagcagc tgatcagagc cgcagagatc
     4801 agggcttccg caaatcttgc tgcaacaaaa atgtccgaat gcgtcttggg ccaatcaaag
     4861 cgcgtcgatt tctgtggaaa gggctaccat ctcatgagct ttccccaaag cgccccgcac
     4921 ggcgtggttt ttctccacgt cacctacgtc cccgctcaag agaagaactt tacaaccgct
     4981 cctgcaattt gccatgatgg gaaagcgcac tttcctcgcg aaggcgtctt tgtctcccag
     5041 ggcacacatt ggtttgttac tcaacggaac ttttatgaac cacaaatcat taccacggac
     5101 aacacgttcg tgtccggcaa ctgtgacgtt gtcatcggaa ttgttcagaa cacggtttac
     5161 gatcctctgc aacccgagct tgactccttt aaggaagaac tggacaagta cttcaaaaat
     5221 cacacctcac cggatgtgga cttgggcgat atctctggca taaatgcgtc agtcgtgaac
     5281 attcaaaagg aaattgatcg actcaacgag gtcgcaaaga atcttaatga atccctgata
     5341 gatctgcagg agttggggaa gtacgagcag tatataaagt ggccttggta tatatggctt
     5401 ggctttatcg ccggcctcat tgcaatcgtg atggtcacca tcatgctgtg ctgcatgacc
     5461 tcatgttgct cctgtcttaa ggggtgctgc tcctgcggtt cctgttgcaa atttgacgag
     5521 gacgacagcg aaccggtgct taagggtgtg aagctgcatt acaccggagg cgattacaag
     5581 gatgacgatg acaagtgacg gccgcctgca ggttaattaa gatctttttc cctctgccaa
     5641 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
     5701 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     5761 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
     5821 ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca
     5881 gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt
     5941 tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt
     6001 actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt
     6061 atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg
     6121 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     6181 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     6241 ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat
     6301 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     6361 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     6421 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac
     6481 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     6541 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     6601 catgtctgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6661 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6721 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6781 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6841 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6901 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6961 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     7021 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     7081 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7141 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7201 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7261 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     7321 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7381 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7441 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7501 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7561 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7621 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7681 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7741 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7801 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7861 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7921 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7981 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     8041 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     8101 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     8161 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8221 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8281 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8341 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8401 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8461 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8521 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8581 ttccccgaaa agtgccacct gg
//
