LOCUS       sequence-327669-        6720 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Human expression vector encoding SARS CoV-2 receptor binding domain 
            (RBD) (amino acids 319-541), containing biotinylation site and 6His 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6720)
  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 6720)
  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: "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 RBD (bio-His)
FEATURES             Location/Qualifiers
     source          1..6720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        42..421
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        422..625
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     576..596
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     622..646
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1898..1918
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1919..1936
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1988..2043
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2042..2061)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4279..4297)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4365..4469
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4470..4538,4539..5330)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4688..4707)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5501..6089
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5990..6009
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6243..6260
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6377..6398
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6413..6418,6419..6436,6437..6443)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6451..6467
                     /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)."
     primer_bind     6456..6478
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6475..6491
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6475..6491
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gtacatttat attggctcat gtccaatatg accgccatgt tgacattgat tattgactag
       61 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      121 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      181 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      241 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      301 tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      361 gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      421 ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt
      481 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      541 ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg
      601 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcctca ctctcttccg
      661 catcgctgtc tgcgagggcc agctgttggg ctcgcggttg aggacaaact cttcgcggtc
      721 tttccagtac tcttggatcg gaaacccgtc ggcctccgaa cggtactccg ccaccgaggg
      781 acctgagcga gtccgcatcg accggatcgg aaaacctctc gagaaaggcg tctaaccagt
      841 cacagtcgca aggtaggctg agcaccgtgg cgggcggcag cgggtggcgg tcggggttgt
      901 ttctggcgga ggtgctgctg atgatgtaat taaagtaggc ggtcttgaga cggcggatgg
      961 tcgaggtgag gtgtggcagg cttgagatcc agctgttggg gtgagtactc cctctcaaaa
     1021 gcgggcatta cttctgcgct aagattgtca gtttccaaaa acgaggagga tttgatattc
     1081 acctggcccg atctggccat acacttgagt gacaatgttc gtgtttctgg tgctgctgcc
     1141 tctggtgtcc agccagcggg tgcagcccac cgaatccatc gtgcggttcc ccaatatcac
     1201 caatctgtgc cccttcggcg aggtgttcaa tgccaccaga ttcgcctctg tgtacgcctg
     1261 gaaccggaag cggatcagca attgcgtggc cgactactcc gtgctgtaca actccgccag
     1321 cttcagcacc ttcaagtgct acggcgtgtc ccctaccaag ctgaacgacc tgtgcttcac
     1381 aaacgtgtac gccgacagct tcgtgatccg gggagatgaa gtgcggcaga ttgcccctgg
     1441 acagacaggc aagatcgccg actacaacta caagctgccc gacgacttca ccggctgtgt
     1501 gattgcctgg aacagcaaca acctggactc caaagtcggc ggcaactaca attacctgta
     1561 ccggctgttc cggaagtcca atctgaagcc cttcgagcgg gacatctcca ccgagatcta
     1621 tcaggccggc agcacccctt gtaacggcgt ggaaggcttc aactgctact tcccactgca
     1681 gtcctacggc tttcagccca caaatggcgt gggctatcag ccctacagag tggtggtgct
     1741 gagcttcgaa ctgctgcatg cccctgccac agtgtgcggc cctaagaaaa gcaccaatct
     1801 cgtgaagaac aaatgcgtga acttcggagg cggtggatcc aattccggat cactgcatca
     1861 tattctggat gcacagaaaa tggtgtggaa tcatcgtgaa aacttgtact ttcaaggcca
     1921 tcaccatcac catcactaag tctagacacg tgattaatta aggatccccc gacctcgacc
     1981 tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc
     2041 tcactcggaa ggacatatgg gagggcaaat catttggtcg agatccctcg gagatctcta
     2101 gctagaggat cgatccccgc cccggacgaa ctaaacctga ctacgacatc tctgcccctt
     2161 cttcgcgggg cagtgcatgt aatcccttca gttggttggt acaacttgcc aactgggccc
     2221 tgttccacat gtgacacggg gggggaccaa acacaaaggg gttctctgac tgtagttgac
     2281 atccttataa atggatgtgc acatttgcca acactgagtg gctttcatcc tggagcagac
     2341 tttgcagtct gtggactgca acacaacatt gcctttatgt gtaactcttg gctgaagctc
     2401 ttacaccaat gctgggggac atgtacctcc caggggccca ggaagactac gggaggctac
     2461 accaacgtca atcagagggg cctgtgtagc taccgataag cggaccctca agagggcatt
     2521 agcaatagtg tttataaggc ccccttgtta accctaaacg ggtagcatat gcttcccggg
     2581 tagtagtata tactatccag actaacccta attcaatagc atatgttacc caacgggaag
     2641 catatgctat cgaattaggg ttagtaaaag ggtcctaagg aacagcgata tctcccaccc
     2701 catgagctgt cacggtttta tttacatggg gtcaggattc cacgagggta gtgaaccatt
     2761 ttagtcacaa gggcagtggc tgaagatcaa ggagcgggca gtgaactctc ctgaatcttc
     2821 gcctgcttct tcattctcct tcgtttagct aatagaataa ctgctgagtt gtgaacagta
     2881 aggtgtatgt gaggtgctcg aaaacaaggt ttcaggtgac gcccccagaa taaaatttgg
     2941 acggggggtt cagtggtggc attgtgctat gacaccaata taaccctcac aaaccccttg
     3001 ggcaataaat actagtgtag gaatgaaaca ttctgaatat ctttaacaat agaaatccat
     3061 ggggtgggga caagccgtaa agactggatg tccatctcac acgaatttat ggctatgggc
     3121 aacacataat cctagtgcaa tatgatactg gggttattaa gatgtgtccc aggcagggac
     3181 caagacaggt gaaccatgtt gttacactct atttgtaaca aggggaaaga gagtggacgc
     3241 cgacagcagc ggactccact ggttgtctct aacacccccg aaaattaaac ggggctccac
     3301 gccaatgggg cccataaaca aagacaagtg gccactcttt tttttgaaat tgtggagtgg
     3361 gggcacgcgt cagcccccac acgccgccct gcggttttgg actgtaaaat aagggtgtaa
     3421 taacttggct gattgtaacc ccgctaacca ctgcggtcaa accacttgcc cacaaaacca
     3481 ctaatggcac cccggggaat acctgcataa gtaggtgggc gggccaagat aggggcgcga
     3541 ttgctgcgat ctggaggaca aattacacac acttgcgcct gagcgccaag cacagggttg
     3601 ttggtcctca tattcacgag gtcgctgaga gcacggtggg ctaatgttgc catgggtagc
     3661 atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatag
     3721 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     3781 gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     3841 gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg ggtagcatat
     3901 gctatcctaa tagagattag ggtagtatat gctatcctaa tttatatctg ggtagcatat
     3961 actacccaaa tatctggata gcatatgcta tcctaatcta tatctgggta gcatatgcta
     4021 tcctaatcta tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta
     4081 tcctaatcta tatctgggta gtatatgcta tcctaattta tatctgggta gcataggcta
     4141 tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta
     4201 tcctaatctg tatccgggta gcatatgcta tcctcatgat aagctgtcaa acatgagaat
     4261 tttcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     4321 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4381 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4441 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4501 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4561 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4621 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4681 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     4741 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4801 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4861 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4921 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4981 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     5041 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5101 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5161 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5221 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5281 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5341 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5401 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5461 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5521 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5581 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5641 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5701 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5761 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5821 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5881 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5941 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6001 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6061 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6121 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6181 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     6241 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     6301 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     6361 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     6421 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     6481 acagctatga ccatgattac gccaagctct agctagaggt cgaccaattc tcatgtttga
     6541 cagcttatca tcgcagatcc gggcaacgtt gttgccattg ctgcaggcgc agaactggta
     6601 ggtatggaag atctatacat tgaatcaata ttggcaatta gccatattag tcattggtta
     6661 tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat
//
