LOCUS       sequence-371279-        9218 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express chimeric SARS-CoV spike with the SARS-CoV-2 receptor binding
            domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9218)
  AUTHORS   Roelle SM, Shukla N, Pham AT, Bruchez AM, Matreyek KA
  TITLE     Expanded ACE2 dependencies of diverse SARS-like coronavirus receptor
            binding domains.
  JOURNAL   PLoS Biol. 2022 Jul 27;20(7):e3001738. doi: 
            10.1371/journal.pbio.3001738.
  PUBMED    35895696
REFERENCE   2  (bases 1 to 9218)
  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 
            Biol."; date: "2022-07-27"; volume: "20(7):e3001738. doi"; pages: " 
            10.1371/journal.pbio.3001738"
COMMENT     Sequence Label: pcDNA3_SARS-CoV-Spike[SARS-CoV-2 RBD]
FEATURES             Location/Qualifiers
     source          1..9218
                     /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(4577..4594)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4583..4807
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4853..5281
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4940..4959)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5150..5171
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5290..5310)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5295..5624
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5475..5610
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5745..5764)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6353..6372
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6657..6778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6694..6713)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6748..6767
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6827..6843)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6827..6843)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6840..6862)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6851..6867
                     /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(6875..6881,6882..6899,6900..6905))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6920..6941
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7058..7075)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7229..7817)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7309..7328)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7988..8779,8780..8848))
                     /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     8611..8630
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8849..8953)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9028..9047)
                     /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 gctagcgttt aaacgggccc tctagaaacc atgcccatgg ggtctctgca accgctggcc
      721 accttgtacc tgctggggat gctggtcgct tccgtgctag ccagcggcag cgacctggac
      781 cgctgcacca cgttcgatga cgtgcaggcg cccaactaca cccagcacac ctccagcatg
      841 cgcggcgtgt actatcccga cgagatcttc cgcagcgaca ccctgtacct cacccaggac
      901 ctgttcctgc ccttctactc caacgtgacc ggcttccaca ccatcaacca caccttcggc
      961 aaccccgtga tcccgttcaa ggacggcatc tacttcgccg cgaccgagaa gagcaacgtg
     1021 gtccgcggct gggtgttcgg gagcaccatg aataacaagt cccagagcgt gatcattatc
     1081 aataactcca ccaacgtcgt gatccgcgcc tgcaacttcg agctgtgtga caacccgttc
     1141 tttgccgtga gcaagcccat gggcacccag acgcacacca tgatcttcga caacgccttc
     1201 aactgcacct tcgagtacat cagcgacgcc ttctccctgg acgtgagcga gaagtccgga
     1261 aacttcaagc acctgcgcga gttcgtgttc aagaacaaag acggcttcct gtacgtgtac
     1321 aagggctacc agcccatcga cgtcgtgcgc gacctgccca gcggcttcaa caccctgaag
     1381 cccatcttca agctgcccct cggcattaac atcaccaact tccgcgccat cctgaccgcc
     1441 ttcagccctg cccaggacat ctggggcacc agcgccgcgg cctacttcgt cggctacctg
     1501 aagcccacga ccttcatgct gaagtacgac gagaacggca cgatcaccga tgccgtggac
     1561 tgcagccaga acccgctcgc cgagctgaag tgcagcgtga agtccttcga gatcgacaag
     1621 ggcatctacc agaccagcaa cttccgggtg cagcccaccg aatccatcgt gcggttcccc
     1681 aatatcacca atctgtgccc cttcggcgag gtgttcaatg ccaccagatt cgcctctgtg
     1741 tacgcctgga accggaagcg gatcagcaat tgcgtggccg actactccgt gctgtacaac
     1801 tccgccagct tcagcacctt caagtgctac ggcgtgtccc ctaccaagct gaacgacctg
     1861 tgcttcacaa acgtgtacgc cgacagcttc gtgatccggg gagatgaagt gcggcagatt
     1921 gcccctggac agacaggcaa gatcgccgac tacaactaca agctgcccga cgacttcacc
     1981 ggctgtgtga ttgcctggaa cagcaacaac ctggactcca aagtcggcgg caactacaat
     2041 tacctgtacc ggctgttccg gaagtccaat ctgaagccct tcgagcggga catctccacc
     2101 gagatctatc aggccggcag caccccttgt aacggcgtgg aaggcttcaa ctgctacttc
     2161 ccactgcagt cctacggctt tcagcccaca aatggcgtgg gctatcagcc ctacagagtg
     2221 gtggtgctga gcttcgaact gctgcatgcc cctgccacag tgtgcggccc taagaaaagc
     2281 accaatctcg tgaagaacaa atgcgtgaac ttcaacttca acggcctgac cggcaccggc
     2341 gtgctgacag agagcaacaa gaagttcctg ccattccagc agttcggccg cgacgtcagc
     2401 gacttcaccg acagcgtgcg cgacccgaag accagcgaga tcctggacat ctctccctgc
     2461 tccttcggcg gtgtcagcgt gatcacgccc ggcaccaacg ccagctccga ggtggccgtc
     2521 ctgtaccagg acgtgaactg caccgacgtc agcaccgcca tccacgcgga ccagctgacg
     2581 cccgcctggc gcatctacag caccggcaac aacgtgttcc agacccaggc cggctgcctg
     2641 atcggcgccg agcacgtcga caccagctac gagtgcgaca tcccgattgg cgccgggatc
     2701 tgcgccagct accacaccgt gagcctgctc cgctccacca gccagaagtc catcgtggcc
     2761 tacaccatga gcctgggcgc cgacagcagc atcgcctaca gcaacaacac catcgccatc
     2821 ccgaccaact tctccatcag catcacgacc gaggtgatgc ccgtgagcat ggccaagacc
     2881 agcgtggact gcaacatgta catctgcggc gacagcaccg agtgcgccaa cctgctcctg
     2941 cagtacggct ccttctgcac ccagctgaac cgcgccctga gcggcatcgc cgccgagcag
     3001 gaccgcaaca cccgcgaggt gttcgcccag gtgaagcaga tgtacaagac gcccaccctg
     3061 aagtacttcg gcggcttcaa cttcagccag atcctgcccg acccgctgaa gcccaccaag
     3121 cgcagcttca tcgaggacct gctcttcaac aaggtgacgc tggccgacgc cggcttcatg
     3181 aagcagtacg gcgagtgcct gggcgacatc aacgcccgcg acctgatctg cgcccagaag
     3241 ttcaacggcc tgaccgtgct cccgccgctg ctgaccgacg acatgatcgc cgcgtacacc
     3301 gccgcgctgg tgagcggcac cgccacggcc ggctggacct tcggcgccgg cgcggccctg
     3361 cagatcccgt tcgccatgca gatggcctac cgcttcaacg gcatcggcgt gacccagaac
     3421 gtgctgtacg agaaccagaa gcagatcgcc aaccagttca acaaggccat cagccagatc
     3481 caggagagcc tgaccacgac cagcacggcc ctgggcaagc tgcaggacgt ggtcaaccag
     3541 aacgcccagg ccctgaacac cctggtgaag cagctgtcca gcaacttcgg cgccatcagc
     3601 tccgtgctga acgatatcct gagccgcctg gacaaggtgg aggccgaggt gcagatcgat
     3661 cgcctgatca ccggccgcct gcagagcctg cagacctacg tgacccagca gctgatccgc
     3721 gccgcggaga tccgcgccag cgcgaacctg gccgccacca agatgtccga gtgcgtgctg
     3781 ggccagagca agcgcgtgga cttctgcggc aagggctacc acctgatgag cttcccgcag
     3841 gccgcgccgc acggcgtggt cttcctgcac gtgacctacg tgcccagcca ggagcgcaac
     3901 ttcaccacgg cgcccgccat ctgccacgag ggcaaggcct acttcccgcg cgagggcgtg
     3961 ttcgtgttca acggcaccag ctggttcatc acccagcgca acttcttcag cccgcaaatc
     4021 atcaccacgg acaacacctt cgtgagcggc aactgcgacg tcgtgatcgg catcatcaac
     4081 aacaccgtgt acgacccgct gcagcccgag ctggactcct tcaaggagga gctggacaag
     4141 tacttcaaga accacaccag cccggacgtg gacctgggcg acatcagcgg catcaacgcc
     4201 agcgtcgtga acatccagaa ggagatcgac cgcctgaacg aggtggccaa gaacctgaac
     4261 gagagcctga tcgacctcca ggagctgggc aagtacgagc agtacatcaa gtggccttgg
     4321 tatgtgtggc tgggcttcat cgccgggctg atcgccatcg tgatggtgac catcctgctc
     4381 tgctgcatga ccagctgctg ctcctgcctg aagggcgcct gcagctgcgg cagctgctgc
     4441 aagttcgacg aggacgacag cgaccccgtg ctgaagggcg tgaagctgca ctacacgggc
     4501 ggcaccgaga ccagccaggt ggcgcccgcc taggatccga gctcggtacc aagcttaagt
     4561 ttaaaccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     4621 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     4681 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     4741 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     4801 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     4861 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4921 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4981 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     5041 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     5101 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     5161 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     5221 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5281 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     5341 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     5401 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     5461 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5521 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     5581 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     5641 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     5701 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     5761 gggcaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5821 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     5881 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5941 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     6001 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     6061 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     6121 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     6181 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     6241 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     6301 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     6361 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     6421 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     6481 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     6541 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     6601 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     6661 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6721 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6781 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     6841 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6901 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6961 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     7021 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     7081 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7141 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7201 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7261 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7321 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7381 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7441 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7501 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7561 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7621 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     7681 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7741 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7801 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7861 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7921 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7981 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     8041 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     8101 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     8161 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     8221 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     8281 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     8341 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     8401 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     8461 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     8521 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     8581 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8641 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8701 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8761 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8821 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8881 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8941 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     9001 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     9061 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     9121 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     9181 tgcgctgctt cgcgatgtac gggccagata tacgcgtt
//
