LOCUS       sequence-316774-        6037 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for RBD of bat coronavirus isolate 
            Rs4231.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6037)
  AUTHORS   Chan KK, Tan TJC, Narayanan KK, Procko E
  TITLE     An engineered decoy receptor for SARS-CoV-2 broadly binds protein S 
            sequence variants.
  JOURNAL   Sci Adv. 2021 Feb 17;7(8). pii: 7/8/eabf1738. doi: 
            10.1126/sciadv.abf1738. Print 2021 Feb.
  PUBMED    33597251
REFERENCE   2  (bases 1 to 6037)
  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: "Sci Adv."; 
            date: "2021-02-17"; volume: "7(8). pii: 7/8/eabf1738. doi"; pages: "
            10.1126/sciadv.abf1738. Print 2021 Feb"
COMMENT     Sequence Label: pcDNA3-Rs4231-RBD-8h
FEATURES             Location/Qualifiers
     source          1..6037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        117..496
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        497..700
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     651..671
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     745..764
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        745..763
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1443..1466
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(1510..1527)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1516..1740
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1786..2214
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1873..1892)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2083..2104
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2223..2243)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2228..2557
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2408..2543
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2470..2489
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2624..3418
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2678..2697)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3288..3307
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3592..3713
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3629..3648)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3683..3702
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3762..3778)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3762..3778)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3775..3797)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3786..3802
                     /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(3810..3816,3817..3834,3835..3840))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3855..3876
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3993..4010)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4164..4752)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4244..4263)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4923..5714,5715..5783))
                     /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     5546..5565
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5784..5888)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5963..5982)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
       61 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      121 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      181 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      241 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      301 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      361 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      421 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      481 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      541 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      601 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      661 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      721 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      781 gcgccaccat gaagaccatc atcgccctga gctacatctt ctgcctggtg ttcgccacaa
      841 acttgtgtcc gtttggtgag gtatttaacg caacgacttt tccgtcagtc tatgcctggg
      901 agaggaaaag gataagcaac tgtgttgctg attattccgt gctctacaac tccactagtt
      961 tttctacgtt caaatgttat ggggttagcg ccaccaagct taacgacctt tgtttctcaa
     1021 atgtttacgc tgactcattt gttgttaaag gggacgacgt gcggcaaatc gctccaggcc
     1081 agacaggcgt tatcgcggat tacaattaca agctccctga tgatttcctg gggtgtgtac
     1141 tggcgtggaa tactaacagt aaagattcct caactagcgg gaactacaat tatctgtata
     1201 gatgggtaag gcgcagcaaa ctgaaccctt atgaacggga tctgtctaat gacatctaca
     1261 gcccaggggg acaatcttgc tcagcgattg gaccaaactg ctataatccc ttgagacctt
     1321 acgggttttt cactaccgct ggagtcgggc atcagccata ccgggttgtt gtcctttcct
     1381 ttgagctcct taatgcccca gccacggtct gcggccccaa actttccacg gacggatccg
     1441 gtcaccatca ccaccaccat catcattgat aactcgagtc tagagggccc gtttaaaccc
     1501 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     1561 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     1621 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     1681 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     1741 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     1801 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     1861 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     1921 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     1981 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     2041 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2101 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     2161 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     2221 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2281 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     2341 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     2401 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     2461 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     2521 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     2581 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     2641 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     2701 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     2761 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     2821 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     2881 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     2941 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     3001 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     3061 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     3121 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     3181 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     3241 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     3301 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     3361 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     3421 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     3481 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     3541 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     3601 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3661 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     3721 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     3781 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     3841 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     3901 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3961 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4021 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4081 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4141 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4201 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4261 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4321 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4381 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4441 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4501 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4561 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4621 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4681 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4741 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4801 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4861 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4921 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4981 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5041 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5101 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5161 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5221 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5281 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5341 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5401 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5461 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5521 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5581 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5641 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5701 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5761 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5821 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5881 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     5941 atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     6001 tgctccctgc ttgtgtgttg gaggtcgctg agtagtg
//
