LOCUS       sequence-323869-        8331 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human expression vector encoding full length SARS CoV-2 spike 
            protein, containing biotinylation site and 8His-tag. Modification of
            Addgene plasmid #154754..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8331)
  AUTHORS   Reyes RA, Clarke K, Gonzales SJ, Cantwell AM, Garza R, Catano G, 
            Tragus RE, Patterson TF, Bol S, Bunnik EM
  TITLE     SARS-CoV-2 spike-specific memory B cells express higher levels of 
            T-bet and FcRL5 after non-severe COVID-19 as compared to severe 
            disease.
  JOURNAL   PLoS One. 2021 Dec 22;16(12):e0261656. doi: 
            10.1371/journal.pone.0261656. eCollection 2021.
  PUBMED    34936684
REFERENCE   2  (bases 1 to 8331)
  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: "PLoS One.";
            date: "2021-12-22"; volume: "16(12):e0261656. doi"; pages: " 
            10.1371/journal.pone.0261656. eCollection 2021"
COMMENT     Sequence Label: SARS CoV-2 HexaPro spike (bio-His)
FEATURES             Location/Qualifiers
     source          1..8331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     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             5561..5584
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(5686..5705)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5751..5806
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5805..5824)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6167..6183)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6167..6183)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6180..6202)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6191..6207
                     /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(6215..6221,6222..6239,6240..6245))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6260..6281
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6410..6427)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6581..7169)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6661..6680)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7340..8131,8132..8200))
                     /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     7963..7982
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8201..8305)
                     /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 gggggggcgc gccrgggggg gggggggggg gggggggggg
      541 ggggggggsg rggsggagag gkgcggcggc 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 rgggctcggg 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 ccacccgaac ccagctccca ccagcctaca ccaacagctt
     1861 tacacggggc gtgtactacc ctgacaaggt gttcagatct agcgtcctgc acagcactca
     1921 ggacctcttc ctgccgttct tcagcaacgt gacatggttc cacgccatcc acgtgagcgg
     1981 cacaaacgga accaagcggt ttgataaccc cgtcctgcca ttcaatgatg gagtttactt
     2041 cgccagtacc gagaagagta acatcatccg gggctggatc ttcggcacca ccctggatag
     2101 caaaacacag agcctcctga tcgtgaacaa tgccacgaac gtcgtgatca aggtgtgcga
     2161 gttccagttt tgcaatgatc ctttcctggg tgtgtactac cacaagaaca acaagagctg
     2221 gatggaaagc gagttcagag tctacagcag cgccaacaac tgcacattcg agtacgtctc
     2281 tcagcctttt ctgatggacc ttgaggggaa acaaggcaac ttcaagaacc tgagagaatt
     2341 cgtgttcaag aacatcgacg gctacttcaa aatctactcc aagcacacac ccatcaacct
     2401 ggtccgggac ctccctcagg gcttcagcgc cctggaaccc ctggtcgacc tgcccatagg
     2461 catcaacata acgcggttcc aaaccctgct ggccctgcat agatcctacc tgactcctgg
     2521 cgacagcagc agcggatgga ccgccggagc tgcagcctac tatgtgggct acctgcaacc
     2581 tagaaccttc ctgctgaagt acaacgagaa cggcacaatc acagacgccg tcgactgcgc
     2641 cctggaccct ctctctgaga caaagtgcac cctgaagtcc ttcaccgtgg aaaagggcat
     2701 ctaccagacc agcaacttcc gggtgcagcc tacagagagc atcgtgcgat ttccaaacat
     2761 taccaacctc tgccccttcg gcgaggtgtt taacgccaca agatttgcct ccgtttacgc
     2821 ctggaataga aagagaatca gcaattgtgt ggccgactac tccgtgctgt ataacagcgc
     2881 ctctttcagc accttcaagt gctacggcgt ttccccaaca aagctgaatg acctgtgctt
     2941 caccaacgtg tacgccgact ccttcgtaat tagaggcgat gaggtgcggc agatcgcacc
     3001 aggccagacc ggtaagatcg ctgactacaa ctataagctg cctgatgatt ttacaggctg
     3061 cgtgatcgcc tggaactcta acaacctgga tagcaaggtg ggcggcaact acaactacct
     3121 gtaccggctg tttcgcaagt ctaacctgaa acctttcgag agagacatct ccacagagat
     3181 ctaccaggcc ggttctacac cttgtaacgg ggtggaaggc ttcaactgtt acttccctct
     3241 gcaaagctac ggcttccagc ctaccaatgg agtcggctac cagccatacc gggtggtcgt
     3301 gctgtccttc gagttactcc acgcccccgc caccgtctgc ggtcctaaga agtccaccaa
     3361 tctggttaag aacaaatgcg tgaacttcaa cttcaacggc ctgaccggga ccggcgtgct
     3421 gaccgaaagc aacaaaaagt tcctcccctt ccagcagttc ggccgtgata tcgctgacac
     3481 cacagatgcc gtcagagatc cacagaccct ggaaatcctg gatattacac cctgctcctt
     3541 cggaggagtt tctgtgatca cccccgggac caataccagc aaccaggtgg ctgtgctgta
     3601 ccaagatgtt aactgcaccg aggttcctgt ggccatccac gccgatcagc tgacacctac
     3661 ttggagagtg tactccactg gctccaatgt gttccagacc agggccggat gtctgatcgg
     3721 cgccgagcac gtgaataaca gttacgagtg cgacatccct atcggcgccg gcatctgtgc
     3781 cagctaccag acccagacaa acagccctgg gtctgcttcc tctgtagcta gccagagcat
     3841 catcgcctac accatgagcc tgggcgcaga gaacagcgtg gcctattcca acaactctat
     3901 cgccattccc accaacttta caattagcgt cacaacagag atcctgcccg tgagcatgac
     3961 caagaccagc gtggactgta caatgtacat ctgtggcgac agcactgaat gcagcaacct
     4021 gctgctgcaa tacggctcct tttgcaccca actgaaccgg gcgctgaccg gaatcgccgt
     4081 ggaacaggac aaaaataccc aggaggtgtt cgcccaagtg aagcagatct acaagacccc
     4141 acctatcaag gacttcggcg gctttaactt tagccagatt ctccctgatc cttctaagcc
     4201 tagcaagcgg agccctatcg aggatctgct gttcaacaag gtcaccctgg ccgatgccgg
     4261 ctttatcaaa cagtatggcg attgcctggg cgacatagcc gccagagatc tgatctgcgc
     4321 ccagaaattc aacggcctga cagttctccc acctctgctg accgacgaga tgatcgctca
     4381 gtacacctct gccctgctgg ctggcaccat cacatctggg tggacatttg gcgccggccc
     4441 cgccctgcag atcccctttc ccatgcagat ggcctataga ttcaacggaa tcggcgtgac
     4501 ccagaacgtg ctgtatgaaa accagaagct gatcgctaac cagttcaatt ctgccatcgg
     4561 caagatccag gactccctct cctctacccc cagcgccctg ggcaaactgc aggacgtggt
     4621 gaatcagaac gcccaagccc tgaacaccct ggtgaagcag ctcagcagca attttggcgc
     4681 catcagctct gtgctgaacg atatcctgtc tagactggac cctccagaag ccgaagtcca
     4741 gatcgataga ctgatcacag gcagactgca gtccctgcaa acctacgtga cccaacagct
     4801 gatcagggcc gctgaaataa gagccagcgc caatctcgcc gctaccaaga tgtccgagtg
     4861 tgtgctggga cagtctaaac gcgttgactt ctgcggcaaa ggctatcacc tgatgagctt
     4921 cccccagagc gcgccgcacg gcgtggtgtt cctgcatgtg acatacgtgc ctgcccaaga
     4981 gaagaatttc acaaccgccc ctgccatctg ccacgacggc aaggcccact tcccaagaga
     5041 gggcgttttc gtttccaatg gcacacactg gttcgtgaca caaagaaact tctacgaacc
     5101 ccagattatc accaccgaca acaccttcgt gagtggcaat tgtgacgtgg tcatcggaat
     5161 cgtgaacaac acagtgtacg accctctgca acctgagctg gactctttta aggaagagct
     5221 ggacaagtac tttaaaaacc acaccagccc cgatgtggac ctgggcgaca tcagtggcat
     5281 taacgccagc gtggtgaaca tccaaaagga aatcgacaga ctgaacgagg tggccaagaa
     5341 cctgaacgag tccctgatcg acctgcagga gctcggcaaa tacgagcagg gatccggata
     5401 catccccgag gcccccagag atggccaggc ctacgtgcgg aaggacggcg agtgggtact
     5461 gctgagcaca ttcctgggca gatccggagg cggtggttcc ctgcatcata ttctggatgc
     5521 acagaaaatg gtgtggaatc atcgtggcgg tggaggttcc catcaccacc atcaccacca
     5581 tcattgataa tgactcgagc gataattcac tcctcaggtg caggctgcct atcagaaggt
     5641 ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc
     5701 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
     5761 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     5821 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
     5881 tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa
     5941 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
     6001 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
     6061 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
     6121 cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc
     6181 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6241 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6301 ccgctttcca gtcgggaaac ctgtcgtgcc agcggatcga tccgctgcat taatgaatcg
     6361 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6421 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6481 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6541 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6601 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6661 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6721 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6781 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6841 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6901 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6961 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7021 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7081 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7141 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7201 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7261 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7321 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7381 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7441 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7501 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7561 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7621 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7681 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7741 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7801 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7861 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7921 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7981 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8041 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8101 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8161 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8221 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8281 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct g
//
