LOCUS       sequence-423196-        8364 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian cell expression of SARS-CoV-2 Spike protein of 
            Omicron.XBB.1.5 with with furin side intact.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8364)
  AUTHORS   Zhang QE, Lindenberger J, Parsons R, Thakur B, Parks R, Park CS, 
            Huang X, Sammour S, Janowska K, Spence TN, Edwards RJ, Martin M, 
            Williams WB, Gobeil S, Montefiori DC, Korber B, Saunders KO, Haynes 
            BF, Haynes BF, Henderson R, Acharya P
  TITLE     SARS-CoV-2 Omicron XBB lineage spike structures, conformations, 
            antigenicity, and receptor recognition.
  JOURNAL   bioRxiv [Preprint]. 2024 Mar 12:2024.02.12.580004. doi: 
            10.1101/2024.02.12.580004.
  PUBMED    38405707
REFERENCE   2  (bases 1 to 8364)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2024 Mar 12:2024.02.12.580004. doi: 
            10.1101/2024.02.12.580004."
COMMENT     Sequence Label: p-alpha-H-S-RRAR/XBB.1.5
FEATURES             Location/Qualifiers
     source          1..8364
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             5474..5497
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             5501..5524
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             5531..5554
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             5594..5617
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(5719..5738)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5784..5839
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5838..5857)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6200..6216)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6200..6216)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6213..6235)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6224..6240
                     /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(6248..6254,6255..6272,6273..6278))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6293..6314
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6443..6460)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6614..7202)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6694..6713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7373..8164,8165..8233))
                     /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     7996..8015
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8234..8338)
                     /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 gggggsgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagcgcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat tgcggccgtc
     1741 tcaggccgag ttcggtaccg ccaccatgtt cgtgttcctg gtgctcctgc ctctggtgag
     1801 cagccagtgc gtgaacctga tcacccgaac ccagtcctac accaacagct ttacacgggg
     1861 cgtgtactac cctgacaagg tgttcagatc tagcgtcctg cacagcactc aggacctctt
     1921 cctgccgttc ttcagcaacg tgacatggtt ccacgccatc cacgtgagcg gcacaaacgg
     1981 aaccaagcgg tttgataacc ccgccctgcc attcaatgat ggagtttact tcgccagtac
     2041 cgagaagagt aacatcatcc ggggctggat cttcggcacc accctggata gcaaaacaca
     2101 gagcctcctg atcgtgaaca atgccacgaa cgtcgtgatc aaggtgtgcg agttccagtt
     2161 ttgcaatgat cctttcctgg acgtgtacca gaagaacaac aagagctgga tggaaagcga
     2221 gttcagagtc tacagcagcg ccaacaactg cacattcgag tacgtctctc agccttttct
     2281 gatggacctt gaggggaaag aaggcaactt caagaacctg agagaattcg tgttcaagaa
     2341 catcgacggc tacttcaaaa tctactccaa gcacacaccc atcaacctgg agcgggacct
     2401 ccctcagggc ttcagcgccc tggaacccct ggtcgacctg cccataggca tcaacataac
     2461 gcggttccaa accctgctgg ccctgcatag atcctacctg actcctgtgg acagcagcag
     2521 cggatggacc gccggagctg cagcctacta tgtgggctac ctgcaaccta gaaccttcct
     2581 gctgaagtac aacgagaacg gcacaatcac agacgccgtc gactgcgccc tggaccctct
     2641 ctctgagaca aagtgcaccc tgaagtcctt caccgtggaa aagggcatct accagaccag
     2701 caacttccgg gtgcagccta cagagagcat cgtgcgattt ccaaacatta ccaacctctg
     2761 ccccttccac gaggtgttta acgccacaac atttgcctcc gtttacgcct ggaatagaaa
     2821 gagaatcagc aattgtgtgg ccgactactc cgtgatctat aacttcgccc ctttcttcgc
     2881 cttcaagtgc tacggcgttt ccccaacaaa gctgaatgac ctgtgcttca ccaacgtgta
     2941 cgccgactcc ttcgtaatta gaggcaatga ggtgtcccag atcgcaccag gccagaccgg
     3001 taacatcgct gactacaact ataagctgcc tgatgatttt acaggctgcg tgatcgcctg
     3061 gaactccaac aagctggata gcaagcctag cggcaactac aactacctgt accggctgtt
     3121 tcgcaagtct aagctgaaac ctttcgagag agacatctcc acagagatct accaggccgg
     3181 taacaagcct tgtaacgggg tggccggccc caactgttac tcccctctgc aaagctacgg
     3241 cttccggcct acctatggag tcggccacca gccataccgg gtggtcgtgc tgtccttcga
     3301 gttactccac gcccccgcca ccgtctgcgg tcctaagaag tccaccaatc tggttaagaa
     3361 caaatgcgtg aacttcaact tcaacggcct gaccgggacc ggcgtgctga ccgaaagcaa
     3421 caaaaagttc ctccccttcc agcagttcgg ccgtgatatc gctgacacca cagatgccgt
     3481 cagagatcca cagaccctgg aaatcctgga tattacaccc tgctccttcg gaggagtttc
     3541 tgtgatcacc cccgggacca ataccagcaa ccaggtggct gtgctgtacc aaggtgttaa
     3601 ctgcaccgag gttcctgtgg ccatccacgc cgatcagctg acacctactt ggagagtgta
     3661 ctccactggc tccaatgtgt tccagaccag ggccggatgt ctgatcggcg ccgagtacgt
     3721 gaataacagt tacgagtgcg acatccctat cggcgccggc atctgtgcca gctaccagac
     3781 ccagacaaag agccatagac gggctagatc tgtagctagc cagagcatca tcgcctacac
     3841 catgagcctg ggcgcagaga acagcgtggc ctattccaac aactctatcg ccattcccac
     3901 caactttaca attagcgtca caacagagat cctgcccgtg agcatgacca agaccagcgt
     3961 ggactgtaca atgtacatct gtggcgacag cactgaatgc agcaacctgc tgctgcaata
     4021 cggctccttt tgcacccaac tgaagcgggc gctgaccgga atcgccgtgg aacaggacaa
     4081 aaatacccag gaggtgttcg cccaagtgaa gcagatctac aagaccccac ctatcaagta
     4141 cttcggcggc tttaacttta gccagattct ccctgatcct tctaagccta gcaagcggag
     4201 ctttatcgag gatctgctgt tcaacaaggt caccctggcc gatgccggct ttatcaaaca
     4261 gtatggcgat tgtctgggcg acatagccgc cagagatctg atctgcgccc agaaattcaa
     4321 cggcctgaca gttctcccac ctctgctgac cgacgagatg atcgctcagt acacctctgc
     4381 cctgctggct ggcaccatca catctgggtg gacatttggc gccggcgccg ccctgcagat
     4441 cccctttgcc atgcagatgg cctatagatt caacggaatc ggcgtgaccc agaacgtgct
     4501 gtatgaaaac cagaagctga tcgctaacca gttcaattct gccatcggca agatccagga
     4561 ctccctctcc tctaccgcca gcgccctggg caaactgcag gacgtggtga atcacaacgc
     4621 ccaagccctg aacaccctgg tgaagcagct cagcagcaag tttggcgcca tcagctctgt
     4681 gctgaacgat atcctgtcta gactggacaa ggtggaagcc gaagtccaga tcgatagact
     4741 gatcacaggc agactgcagt ccctgcaaac ctacgtgacc caacagctga tcagggccgc
     4801 tgaaataaga gccagcgcca atctcgccgc taccaagatg tccgagtgtg tgctgggaca
     4861 gtctaaacgc gttgacttct gcggcaaagg ctatcacctg atgagcttcc cccagagcgc
     4921 gccgcacggc gtggtgttcc tgcatgtgac atacgtgcct gcccaagaga agaatttcac
     4981 aaccgcccct gccatctgcc acgacggcaa ggcccacttc ccaagagagg gcgttttcgt
     5041 ttccaatggc acacactggt tcgtgacaca aagaaacttc tacgaacccc agattatcac
     5101 caccgacaac accttcgtga gtggcaattg tgacgtggtc atcggaatcg tgaacaacac
     5161 agtgtacgac cctctgcaac ctgagctgga ctcttttaag gaagagctgg acaagtactt
     5221 taaaaaccac accagccccg atgtggacct gggcgacatc agtggcatta acgccagcgt
     5281 ggtgaacatc caaaaggaaa tcgacagact gaacgaggtg gccaagaacc tgaacgagtc
     5341 cctgatcgac ctgcaggagc tcggcaaata cgagcaggga tccggataca tccccgaggc
     5401 ccccagagat ggccaggcct acgtgcggaa ggacggcgag tgggtactgc tgagcacatt
     5461 cctgggcaga tccctggagg tgctgttcca gggcccaggc catcaccacc atcaccacca
     5521 tcatagcgcc tggtcccacc cccagttcga gaagggcggc ggtagtggag ggggcggatc
     5581 tggcggctca gcttggagcc acccccagtt cgaaaagtga taatgactcg agcgataatt
     5641 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     5701 tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc
     5761 ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     5821 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca
     5881 gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa
     5941 aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt
     6001 ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt
     6061 ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc
     6121 ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc
     6181 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6241 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6301 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6361 gccagcggat cgatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6421 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6481 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6541 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6601 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6661 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6721 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6781 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6841 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6901 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6961 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7021 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7081 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7141 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7201 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7261 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7321 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7381 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7441 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7501 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7561 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7621 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7681 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7741 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7801 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7861 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7921 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7981 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8041 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8101 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8161 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8221 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8281 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8341 catttccccg aaaagtgcca cctg
//
