LOCUS       sequence-371274-        9215 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express chimeric SARS-CoV spike with the BtKY72 receptor binding 
            domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9215)
  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 9215)
  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[BtKY72 RBD]
FEATURES             Location/Qualifiers
     source          1..9215
                     /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(4574..4591)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4580..4804
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4850..5278
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4937..4956)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5147..5168
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5287..5307)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5292..5621
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5472..5607
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5742..5761)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6350..6369
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6654..6775
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6691..6710)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6745..6764
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6824..6840)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6824..6840)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6837..6859)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6848..6864
                     /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(6872..6878,6879..6896,6897..6902))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6917..6938
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7055..7072)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7226..7814)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7306..7325)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7985..8776,8777..8845))
                     /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     8608..8627
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8846..8950)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9025..9044)
                     /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 tgatcgccca
      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 cttccgcgtg agtccttcca ctgaagtggt ccggttcccg
     1681 aacataacta atttgtgccc tttcggccag gtatttaacg cgtcaaattt cccatctgtt
     1741 tacgcctggg aaagattgcg cataagtgac tgtgtcgcag actatgctgt gttgtacaac
     1801 agctccagca gctttagtac atttaagtgc tatggggtat caccaaccaa attgaatgat
     1861 ctctgcttct catccgttta cgctgattac ttcgtcgtta agggggatga cgtgaggcag
     1921 atcgccccgg cgcaaacagg agtaattgcg gattacaact acaaattgcc ggacgatttc
     1981 actggttgtg tacttgcctg gaataccaat agcgttgact ccaaatccgg gaataatttt
     2041 tactaccgac ttttccgaca cggcaagata aagccatatg aacgggatat tagcaacgtc
     2101 ctctataact ctgctggggg gacctgctcc tcaattagcc aactggggtg ctatgagcct
     2161 cttaaatcct acggatttac ccctactgtc ggagttggct accaacccta cagggtagtc
     2221 gtcctctcat tcgagctttt gaatgcgccg gcaactgtat gtggaccaaa gaagagcaca
     2281 gagctggtca agaacaagtg cgtgaacttc aatttcaacg gcctgaccgg caccggcgtg
     2341 ctgacgccca gctccaagcg cttccagccc ttccagcagt tcggccgcga cgtcagcgac
     2401 ttcaccgaca gcgtgcgcga cccgaagacc agcgagatcc tggacatctc tccctgctcc
     2461 ttcggcggtg tcagcgtgat cacgcccggc accaacgcca gctccgaggt ggccgtcctg
     2521 taccaggacg tgaactgcac cgacgtcagc accgccatcc acgcggacca gctgacgccc
     2581 gcctggcgca tctacagcac cggcaacaac gtgttccaga cccaggccgg ctgcctgatc
     2641 ggcgccgagc acgtcgacac cagctacgag tgcgacatcc cgattggcgc cgggatctgc
     2701 gccagctacc acaccgtgag cctgctccgc tccaccagcc agaagtccat cgtggcctac
     2761 accatgagcc tgggcgccga cagcagcatc gcctacagca acaacaccat cgccatcccg
     2821 accaacttct ccatcagcat cacgaccgag gtgatgcccg tgagcatggc caagaccagc
     2881 gtggactgca acatgtacat ctgcggcgac agcaccgagt gcgccaacct gctcctgcag
     2941 tacggctcct tctgcaccca gctgaaccgc gccctgagcg gcatcgccgc cgagcaggac
     3001 cgcaacaccc gcgaggtgtt cgcccaggtg aagcagatgt acaagacgcc caccctgaag
     3061 tacttcggcg gcttcaactt cagccagatc ctgcccgacc cgctgaagcc caccaagcgc
     3121 agcttcatcg aggacctgct cttcaacaag gtgacgctgg ccgacgccgg cttcatgaag
     3181 cagtacggcg agtgcctggg cgacatcaac gcccgcgacc tgatctgcgc ccagaagttc
     3241 aacggcctga ccgtgctccc gccgctgctg accgacgaca tgatcgccgc gtacaccgcc
     3301 gcgctggtga gcggcaccgc cacggccggc tggaccttcg gcgccggcgc ggccctgcag
     3361 atcccgttcg ccatgcagat ggcctaccgc ttcaacggca tcggcgtgac ccagaacgtg
     3421 ctgtacgaga accagaagca gatcaccaac cagttcaaca aggccatcag ccagatccag
     3481 gagagcctga ccacgaccag cacggccctg ggcaagctgc aggacgtggt caaccagaac
     3541 gcccaggccc tgaacaccct ggtgaagcag ctgtccagca acttcggcgc catcagctcc
     3601 gtgctgaacg atatcctgag ccgcctggac aaggtggagg ccgaggtgca gatcgatcgc
     3661 ctgatcaccg gccgcctgca gagcctgcag acctacgtga cccagcagct gatccgcgcc
     3721 gcggagatcc gcgccagcgc gaacctggcc gccaccaaga tgtccgagtg cgtgctgggc
     3781 cagagcaagc gcgtggactt ctgcggcaag ggctaccacc tgatgagctt cccgcaggcc
     3841 gcgccgcacg gcgtggtctt cctgcacgtg acctacgtgc ccagccagga gcgcaacttc
     3901 accacggcgc ccgccatctg ccacgagggc aaggcctact tcccgcgcga gggcgtgttc
     3961 gtgttcaacg gcaccagctg gttcatcacc cagcgcaact tcttcagccc gcaaatcatc
     4021 accacggaca acaccttcgt gagcggcaac tgcgacgtcg tgatcggcat catcaacaac
     4081 accgtgtacg acccgctgca gcccgagctg gactccttca aggaggagct ggacaagtac
     4141 ttcaagaacc acaccagccc ggacgtggac ctgggcgaca tcagcggcat caacgccagc
     4201 gtcgtgaaca tccagaagga gatcgaccgc ctgaacgagg tggccaagaa cctgaacgag
     4261 agcctgatcg acctccagga gctgggcaag tacgagcagt acatcaagtg gccttggtat
     4321 gtgtggctgg gcttcatcgc cgggctgatc gccatcgtga tggtgaccat cctgctctgc
     4381 tgcatgacca gctgctgctc ctgcctgaag ggcgcctgca gctgcggcag ctgctgcaag
     4441 ttcgacgagg acgacagcga ccccgtgctg aagggcgtga agctgcacta cacgggcggc
     4501 accgagacca gccaggtggc gcccgcctag gatccgagct cggtaccaag cttaagttta
     4561 aaccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     4621 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     4681 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     4741 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     4801 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     4861 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4921 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4981 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     5041 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     5101 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     5161 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     5221 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     5281 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5341 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     5401 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     5461 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5521 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     5581 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     5641 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     5701 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     5761 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5821 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     5881 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5941 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     6001 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     6061 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     6121 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     6181 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     6241 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     6301 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     6361 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     6421 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     6481 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     6541 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     6601 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     6661 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6721 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6781 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     6841 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     6901 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     6961 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     7021 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     7081 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     7141 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     7201 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     7261 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     7321 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     7381 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     7441 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     7501 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     7561 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     7621 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     7681 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     7741 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7801 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7861 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7921 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7981 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     8041 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     8101 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     8161 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     8221 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     8281 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     8341 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     8401 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     8461 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     8521 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     8581 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     8641 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     8701 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8761 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8821 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8881 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8941 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     9001 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     9061 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     9121 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     9181 gctgcttcgc gatgtacgga ccagatatac gcgtt
//
