LOCUS       sequence-371277-        9209 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express chimeric SARS-CoV spike with the Khosta-2 receptor binding 
            domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9209)
  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 9209)
  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[Khosta2 RBD]
FEATURES             Location/Qualifiers
     source          1..9209
                     /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(4568..4585)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4574..4798
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4844..5272
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4931..4950)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5141..5162
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5281..5301)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5286..5615
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5466..5601
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5736..5755)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6344..6363
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6648..6769
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6685..6704)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6739..6758
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6818..6834)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6818..6834)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6831..6853)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6842..6858
                     /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(6866..6872,6873..6890,6891..6896))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6911..6932
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7049..7066)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7220..7808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7300..7319)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7979..8770,8771..8839))
                     /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     8602..8621
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8840..8944)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9019..9038)
                     /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 agccctagtt tggacgtagt tcgattccca
     1681 aatatgacca acatttgtcc tttcgatcaa gtcttcaaca aaacgcaatt ccccagcgta
     1741 tatgcttggg agcgagtaag aatctccgac tgcgtttctg attacactgt cctttataac
     1801 tctagcgcca gttttagtac cttcaaatgc tatggggttt caccgacaaa acttaatgat
     1861 ctttgctttt caggagtata tgcagattac ttcgtcgtca agggcgatca cgtacatcag
     1921 atagccccag gtcaaacggg tgtaatcgcg gattataact ataagctccc ctctgaattt
     1981 gtcggatgta tcctggcttg gaatacacga accatagact ctaaacgagg gttctactac
     2041 agactttttc ggcatgggaa tatacgaccg tatgaacggg atacttccaa cgtcccatac
     2101 aatgctgctg gcgggacgtg caatcaacct gggacgcaca actgttacga acccctccag
     2161 gactacgggt tcacctctac ttctggtgtt ggataccagc cgtttcgcgt cgtcgtactt
     2221 tcctttgaac tcctcaatgc tcctgcgaca gtttgcggcc ctaaacagtc aacggacctc
     2281 gtgaaaaaca agtgcgtcaa tttcaatttc aacggcctga ccggcaccgg cgtgctgaca
     2341 gagagcaaca agaagttcct gccattccag cagttcggcc gcgacgtcag cgacttcacc
     2401 gacagcgtgc gcgacccgaa gaccagcgag atcctggaca tctctccctg ctccttcggc
     2461 ggtgtcagcg tgatcacgcc cggcaccaac gccagctccg aggtggccgt cctgtaccag
     2521 gacgtgaact gcaccgacgt cagcaccgcc atccacgcgg accagctgac gcccgcctgg
     2581 cgcatctaca gcaccggcaa caacgtgttc cagacccagg ccggctgcct gatcggcgcc
     2641 gagcacgtcg acaccagcta cgagtgcgac atcccgattg gcgccgggat ctgcgccagc
     2701 taccacaccg tgagcctgct ccgctccacc agccagaagt ccatcgtggc ctacaccatg
     2761 agcctgggcg ccgacagcag catcgcctac agcaacaaca ccatcgccat cccgaccaac
     2821 ttctccatca gcatcacgac cgaggtgatg cccgtgagca tggccaagac cagcgtggac
     2881 tgcaacatgt acatctgcgg cgacagcacc gagtgcgcca acctgctcct gcagtacggc
     2941 tccttctgca cccagctgaa ccgcgccctg agcggcatcg ccgccgagca ggaccgcaac
     3001 acccgcgagg tgttcgccca ggtgaagcag atgtacaaga cgcccaccct gaagtacttc
     3061 ggcggcttca acttcagcca gatcctgccc gacccgctga agcccaccaa gcgcagcttc
     3121 atcgaggacc tgctcttcaa caaggtgacg ctggccgacg ccggcttcat gaagcagtac
     3181 ggcgagtgcc tgggcgacat caacgcccgc gacctgatct gcgcccagaa gttcaacggc
     3241 ctgaccgtgc tcccgccgct gctgaccgac gacatgatcg ccgcgtacac cgccgcgctg
     3301 gtgagcggca ccgccacggc cggctggacc ttcggcgccg gcgcggccct gcagatcccg
     3361 ttcgccatgc agatggccta ccgcttcaac ggcatcggcg tgacccagaa cgtgctgtac
     3421 gagaaccaga agcagatcgc caaccagttc aacaaggcca tcagccagat ccaggagagc
     3481 ctgaccacga ccagcacggc cctgggcaag ctgcaggacg tggtcaacca gaacgcccag
     3541 gccctgaaca ccctggtgaa gcagctgtcc agcaacttcg gcgccatcag ctccgtgctg
     3601 aacgatatcc tgagccgcct ggacaaggtg gaggccgagg tgcagatcga tcgcctgatc
     3661 accggccgcc tgcagagcct gcagacctac gtgacccagc agctgatccg cgccgcggag
     3721 atccgcgcca gcgcgaacct ggccgccacc aagatgtccg agtgcgtgct gggccagagc
     3781 aagcgcgtgg acttctgcgg caagggctac cacctgatga gcttcccgca ggccgcgccg
     3841 cacggcgtgg tcttcctgca cgtgacctac gtgcccagcc aggagcgcaa cttcaccacg
     3901 gcgcccgcca tctgccacga gggcaaggcc tacttcccgc gcgagggcgt gttcgtgttc
     3961 aacggcacca gctggttcat cacccagcgc aacttcttca gcccgcaaat catcaccacg
     4021 gacaacacct tcgtgagcgg caactgcgac gtcgtgatcg gcatcatcaa caacaccgtg
     4081 tacgacccgc tgcagcccga gctggactcc ttcaaggagg agctggacaa gtacttcaag
     4141 aaccacacca gcccggacgt ggacctgggc gacatcagcg gcatcaacgc cagcgtcgtg
     4201 aacatccaga aggagatcga ccgcctgaac gaggtggcca agaacctgaa cgagagcctg
     4261 atcgacctcc aggagctggg caagtacgag cagtacatca agtggccttg gtatgtgtgg
     4321 ctgggcttca tcgccgggct gatcgccatc gtgatggtga ccatcctgct ctgctgcatg
     4381 accagctgct gctcctgcct gaagggcgcc tgcagctgcg gcagctgctg caagttcgac
     4441 gaggacgaca gcgaccccgt gctgaagggc gtgaagctgc actacacggg cggcaccgag
     4501 accagccagg tggcgcccgc ctaggatccg agctcggtac caagcttaag tttaaaccgc
     4561 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     4621 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     4681 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     4741 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     4801 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     4861 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4921 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4981 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     5041 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5101 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5161 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     5221 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     5281 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5341 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     5401 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     5461 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5521 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     5581 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     5641 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     5701 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcaacag
     5761 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     5821 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     5881 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     5941 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     6001 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     6061 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     6121 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     6181 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     6241 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     6301 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     6361 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     6421 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     6481 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     6541 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     6601 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     6661 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6721 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     6781 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     6841 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     6901 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     6961 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     7021 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     7081 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     7141 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     7201 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     7261 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     7321 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7381 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     7441 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7501 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7561 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7621 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     7681 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7741 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7801 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7861 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7921 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7981 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     8041 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     8101 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     8161 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     8221 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     8281 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8341 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8401 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8461 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8521 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8581 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8641 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8701 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8761 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8821 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8881 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8941 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     9001 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     9061 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     9121 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
     9181 tcgcgatgta cgggccagat atacgcgtt
//
