LOCUS       sequence-371281-        9221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express BtKY72 spike.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9221)
  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 9221)
  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_BtKY72-Spike
FEATURES             Location/Qualifiers
     source          1..9221
                     /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(4580..4597)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4586..4810
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4856..5284
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4943..4962)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5153..5174
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5293..5313)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5298..5627
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5478..5613
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5748..5767)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6356..6375
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6660..6781
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6697..6716)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6751..6770
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6830..6846)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6830..6846)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6843..6865)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6854..6870
                     /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(6878..6884,6885..6902,6903..6908))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6923..6944
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7061..7078)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7232..7820)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7312..7331)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7991..8782,8783..8851))
                     /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     8614..8633
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8852..8956)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9031..9050)
                     /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 cctttaccac agctcaagag
      781 ggctgtggga tactcagtaa caaatcaaac ccagcactta cacaatactt ctcatcacga
      841 cgcggctttt attattttga cgatactttt aggtcaagcg ttagagtgtt gacgactggg
      901 tactttctcc catttaacag caacctcaca ggttactcta gccgcaacag cgtaacaggg
      961 cgactcatcc aatttgataa tcctaacata ccattcaagg acggtcttta tttcgcggct
     1021 actgagcgga gcaatgtaat cagaggttgg atatttggga gtacgttgga taatacgacc
     1081 cagtcagcgg ttttgtttaa caacggcacg cacatcgtaa taaacgtatg taatttttat
     1141 ttttgccaag acccgatgtt ggccgtcgcc aacggcagcc attttaagtc ctgggtcttc
     1201 ctcaatgcga cgaattgtac ctataatcga gttcatggtt tcgagatcga cccatcaccg
     1261 aacacaggca gctttatcca ccttcgagag cacgtgttcc ggaacgtgga tgggtttctg
     1321 tatgtatacc acaactacga gcgagttgat gtatatgaca attttcccca aggtttctct
     1381 gtactgaaac cgatttttaa gctcccattc ggactcaata ttacccagtt caaagttatc
     1441 atgaccctct ttagcccaac tacgtcatcc tttaatgctg acgcctcagt atactttgtt
     1501 ggtcatctca agccattgac gatgctcgct gagtttgacg agaacgggac gatcacggat
     1561 gccgttgatt gctcacagga cccgctctct gaacttaaat gtacaacaaa aagtctcaca
     1621 gtagagaagg gaatctatca aaccagtaac tttcgagtca gcccatccac cgaagtggtc
     1681 cggttcccga acataactaa tttgtgccct ttcggccagg tatttaacgc gtcaaatttc
     1741 ccatctgttt acgcctggga aagattgcgc ataagtgact gtgtcgcaga ctatgctgtg
     1801 ttgtacaaca gctccagcag ctttagtaca tttaagtgct atggggtatc accaaccaaa
     1861 ttgaatgatc tctgcttctc atccgtttac gctgattact tcgtcgttaa gggggatgac
     1921 gtgaggcaga tcgccccggc gcaaacagga gtaattgcgg attacaacta caaattgccg
     1981 gacgatttca ctggttgtgt acttgcctgg aataccaata gcgttgactc caaatccggg
     2041 aataattttt actaccgact tttccgacac ggcaagataa agccatatga acgggatatt
     2101 agcaacgtcc tctataactc tgctgggggg acctgctcct caattagcca actggggtgc
     2161 tatgagcctc ttaaatccta cggatttacc cctactgtcg gagttggcta ccaaccctac
     2221 agggtagtcg tcctctcatt cgagcttttg aatgcgccgg caactgtatg tggaccaaag
     2281 aagagcacag agctggtcaa gaacaagtgc gtgaacttca atttcaacgg cctgaccggc
     2341 accggcgtgc tgacgtctag cactaagaaa tttcagccat tccagcaatt tggtagagac
     2401 gtcagcgatt ttacggactc agtaagggat cctaagactt tcgagatcct tgacatttcc
     2461 ccctgttcct atggtggggt ttcagttatt acccccggca ccaacacctc caaagcagtt
     2521 gcagtcctgt accaggacgt aaactgtacc gatgtgccca cgatgataca tgtggagcag
     2581 gttagctccg attggagggt ctacgccttc aatagctatg gtaatatgtt ccaaacccaa
     2641 gcgggctgtc tcgttggtgc gatctacgag aacacaactt acgagtgtga tattccaatc
     2701 ggtgctggaa tttgcgcgaa gttcggaagc gataaaataa ggatgggaca ggaaagtata
     2761 gttgcgtaca ctatgtctat cggagaagat caatcaatcg cgtattctaa taatataata
     2821 gcaattccga ccaatttctc cattagcgta actaccgaag tattgccggt gtctatgacc
     2881 aaaacgtctg tcgactgtaa tatgtacata tgcggagact ccaccgagtg cagtaatttg
     2941 ctgcttcagt atggatcatt ctgcacacaa ctcaacagag cgctgagtgg tattgcggta
     3001 gaacaggacc ggaataccag agacgtattc gcgcagacta aaagtatata taaaacccca
     3061 aatatcaaag acttcggtgg attcaacttc agccaaatcc ttccggatcc gaaaaagctc
     3121 tcatatcgga gcttcatcga agaccttctc tacaacaagg tcacattgtc agatcccggg
     3181 tttatgaagc aatacggaga ctgtctgggc ggtataaatg cgcgcgattt gatttgcgcc
     3241 cagaagttca atggtcttac cgtattgccc cccctcttga ctgacgatat gatagcagca
     3301 tacaccgccg cgttgatttc agggactgca acggctggct atacttttgg ggcaggcgcg
     3361 gccctgcaga tcccgttcgc catgcagatg gcctaccgct tcaacggcat cggcgtgacc
     3421 cagaacgtgc tgtacgagaa ccagaagcag atcgccaacc agttcaacaa cgccataagc
     3481 aagatacagg attcccttac tacaacgtct gcggcattgg gtaagctgca ggatgttatc
     3541 aaccaaaatg cagttgccct gaacaccctg gtgaagcagc tgtccagcaa cttcggcgcc
     3601 atcagctccg tgctgaacga tatcctgagc cgcctggaca aggtggaggc cgaggtgcag
     3661 atcgatcgcc tgatcaccgg ccgcctgcag agcctgcaga cctacgtgac ccagcagctg
     3721 atccgcgccg cggagatccg cgccagcgcg aacctggccg ccaccaagat gtccgagtgc
     3781 gtgctgggcc agagcaagcg cgtggacttc tgcggcaagg gctaccacct gatgagcttc
     3841 ccgcaggccg cgccgcacgg cgtggtcttc ctgcacgtga cctacgtgcc cagccagcag
     3901 cagaacttta ccactgcgcc cgcaatttgc cataacggga aggcgtattt cccgcgggaa
     3961 ggagtatttg taatgaacgg tactcattgg tttattaccc agcggaattt ttatagtcca
     4021 caggtcatta ccacagataa tacctttgaa tcagggtcct gcgatgtggt tataggtatc
     4081 atcaataaca cggtctacga tcccttgcag ccggaactgg aatcttttaa gcaagagttg
     4141 gataaatatt tcaagaatca cacgtctcct gacgtggatt ttggggatat ttcaggtatc
     4201 aatgcttccg ttgttgacat taagaaagaa atagcgcacc tcaacgagat agccaaaaac
     4261 ctgaacgaat ccctgataga tctccaggaa ttggggaaat acgaacagta tgtaaaatgg
     4321 ccttggtacg tttggctggg tttcgtcgct gggatcgtgg ctatcttgat gagcgttata
     4381 atgctctgtt gtatgaccaa ctgctgtagc tgtttcaagg gtatgtgcag ttgtgggagt
     4441 tgttgcaagt tcgacgagga tcacagcgag cctgttagtg gcgtgaagct gcactacacg
     4501 ggcggcaccg agaccagcca ggtggcgccc gcctaggatc cgagctcggt accaagctta
     4561 agtttaaacc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     4621 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     4681 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     4741 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     4801 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     4861 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4921 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4981 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5041 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     5101 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5161 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     5221 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5281 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     5341 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     5401 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     5461 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     5521 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     5581 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     5641 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     5701 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     5761 actgggcaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     5821 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     5881 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5941 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     6001 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     6061 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     6121 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     6181 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     6241 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     6301 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     6361 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     6421 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     6481 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     6541 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     6601 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     6661 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     6721 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     6781 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     6841 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6901 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6961 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     7021 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     7081 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     7141 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     7201 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     7261 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     7321 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     7381 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     7441 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7501 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7561 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7621 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     7681 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7741 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7801 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7861 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7921 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7981 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     8041 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     8101 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     8161 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     8221 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     8281 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     8341 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     8401 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     8461 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8521 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8581 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8641 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8701 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8761 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8821 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8881 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8941 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     9001 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     9061 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     9121 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     9181 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt t
//
