LOCUS       sequence-288058-        6685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for RBD of SARS-CoV-2 protein S (spike)
            fused with IgG1-Fc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6685)
  AUTHORS   Chan KK, Dorosky D, Sharma P, Abbasi SA, Dye JM, Kranz DM, Herbert 
            AS, Procko E
  TITLE     Engineering human ACE2 to optimize binding to the spike protein of 
            SARS coronavirus 2.
  JOURNAL   Science. 2020 Sep 4;369(6508):1261-1265. doi: 
            10.1126/science.abc0870. Epub 2020 Aug 4.
  PUBMED    32753553
REFERENCE   2  (bases 1 to 6685)
  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"; doi: 
            "10.1126/science.abc0870"; journalName: "Science"; date: "2020-09-4-
            4"; volume: "369"; issue: "6508"; pages: "1261-1265"
COMMENT     Sequence Label: pcDNA3-SARS-CoV-2-S-RBD-Fc
FEATURES             Location/Qualifiers
     source          1..6685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2042..2059)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2048..2272
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2318..2746
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2405..2424)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2615..2636
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2755..2775)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2760..3089
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2940..3075
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3002..3021
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3156..3950
                     /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(3210..3229)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3820..3839
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4124..4245
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4161..4180)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4215..4234
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4294..4310)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4294..4310)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4307..4329)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4318..4334
                     /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(4342..4348,4349..4366,4367..4372))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4387..4408
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4525..4542)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4696..5284)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4776..4795)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5455..6246,6247..6315))
                     /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     6078..6097
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6316..6420)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6495..6514)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgcca ccatgaagac catcatcgcc ctgagctaca tcttctgcct ggtgttcgcc
      721 actaatcttt gtccgttcgg tgaggttttt aacgcgacaa ggttcgctag tgtatatgct
      781 tggaaccgaa agagaatctc caattgcgta gctgattact ccgttctcta taacagtgcg
      841 tccttttcaa cctttaagtg ttacggcgtt tctccaacga agctgaatga tctctgtttt
      901 acgaacgtgt atgctgactc tttcgttata cggggggacg aagtgagaca gatagcacca
      961 ggtcagactg ggaagatagc ggattacaac tataagttgc ccgatgattt tacggggtgc
     1021 gtaatcgcat ggaactcaaa caacctcgac tccaaagtag gtggtaatta taattacttg
     1081 tatcgcctgt ttcgaaagag caatttgaag ccttttgagc gggatatttc aaccgaaatt
     1141 taccaagcag gcagtacgcc atgtaacgga gtagagggat ttaattgcta ctttcctctt
     1201 caatcttatg gctttcaacc aacaaacgga gtggggtatc aaccttatag agtggtagta
     1261 ttgtcctttg agctcctcca cgccccggct acagtttgtg ggcccaaaaa gggatccgac
     1321 aaaactcaca catgcccacc gtgcccagca cctgaactcc tggggggacc gtcagtcttc
     1381 ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc
     1441 gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc
     1501 gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag cacgtaccgt
     1561 gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc
     1621 aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa agccaaaggg
     1681 cagccccgag aaccacaggt gtacaccctg cccccatccc gggaggagat gaccaagaac
     1741 caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg
     1801 gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac
     1861 ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcaggggaac
     1921 gtcttctcat gctccgtgat gcacgaggct ctgcacaacc actacacgca gaagagcctc
     1981 tccctgtctc cgggtaaatg ataactcgag tctagagggc ccgtttaaac ccgctgatca
     2041 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2101 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2161 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2221 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     2281 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     2341 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2401 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2461 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2521 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2581 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2641 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2701 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     2761 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2821 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2881 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     2941 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3001 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3061 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3121 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3181 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3241 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3301 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3361 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3421 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3481 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3541 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3601 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3661 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     3721 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3781 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3841 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3901 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     3961 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4021 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4081 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4141 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4201 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     4261 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4321 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4381 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4441 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4501 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4561 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4621 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4681 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4741 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4801 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4861 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4921 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4981 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5041 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5101 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5161 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5221 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5281 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5341 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5401 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5461 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5521 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5581 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5641 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5701 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5761 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5821 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5881 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5941 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6001 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6061 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6121 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6181 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6241 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6301 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6361 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6421 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     6481 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     6541 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     6601 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     6661 gatgtacggg ccagatatac gcgtt
//
