LOCUS       sequence-422363-        5486 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression of SARS-CoV-2 Spike RBD protein of 
            OMICRON.EG5 variant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5486)
  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 5486)
  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: "bioRxiv 
            [Preprint]. 2024 Mar 12:2024.02.12.580004. doi: 
            10.1101/2024.02.12.580004."
COMMENT     Sequence Label: pCAGGS/EG5-RBD
FEATURES             Location/Qualifiers
     source          1..5486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..661
                     /label=chicken beta-actin promoter
     primer_bind     1589..1611
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1673..1692
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2432..2449
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2507..2526)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2572..2627
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2626..2645)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2988..3004)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2988..3004)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3001..3023)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3012..3028
                     /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(3036..3042,3043..3060,3061..3066))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3081..3102
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3161..3356
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3207..3342
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3362..3496
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3399..3418)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3453..3472
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3564..3581)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3735..4323)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3815..3834)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4494..5285,5286..5354))
                     /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     5117..5136
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5355..5459)
                     /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 gggggcgcgc 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 gcgccggcgg ctgtcgaggc
     1321 gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct
     1381 ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg
     1441 cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg
     1501 tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc
     1561 tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct
     1621 agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg
     1681 ctggttattg tgctgtctca tcattttggc aaaggccacc atgttcgtgt ttctggtgct
     1741 gctgcctctg gtgtccagcc agcgggtgca gcctacagag agcatcgtgc gatttccaaa
     1801 cattaccaac ctctgcccct tccacgaggt gtttaacgcc acaacatttg cctccgttta
     1861 cgcctggaat agaaagagaa tcagcaattg tgtggccgac tactccgtga tctataactt
     1921 cgcccctttc ttcgccttca agtgctacgg cgtttcccca acaaagctga atgacctgtg
     1981 cttcaccaac gtgtacgccg actccttcgt aattagaggc aatgaggtgt cccagatcgc
     2041 accaggccag accggtaaca tcgctgacta caactataag ctgcctgatg attttacagg
     2101 ctgcgtgatc gcctggaact ccaacaagct ggatagcaag cctagcggca actacaacta
     2161 cctgtaccgg ctgctgcgca agtctaagct gaaacctttc gagagagaca tctccacaga
     2221 gatctaccag gccggtaaca agccttgtaa cggggtggcc ggccccaact gttactcccc
     2281 tctgcaaagc tacggcttcc ggcctaccta tggagtcggc caccagccat accgggtggt
     2341 cgtgctgtcc ttcgagttac tccacgcccc cgccaccgtc tgcggtccta agaagtccac
     2401 caatctggtt aagaacaaat gcgtgaactt ccaccatcac catcaccatt gataaaattc
     2461 gagctcgcgg ccgcatcgat cttaagtcgc gactcgagct agcagatctt tttccctctg
     2521 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     2581 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     2641 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     2701 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     2761 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     2821 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     2881 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     2941 tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt
     3001 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3061 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3121 cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa
     3181 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3241 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     3301 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3361 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3421 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3481 ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3541 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3601 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3661 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3721 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3781 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3841 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3901 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3961 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4021 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4081 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4141 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4201 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4261 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4321 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4381 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4441 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4501 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4561 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4621 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4681 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4741 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4801 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4861 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4921 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4981 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5041 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5101 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5161 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5221 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5281 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5341 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5401 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5461 acatttcccc gaaaagtgcc acctgg
//
