LOCUS       sequence-444395-        6779 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast surface display of the SARS-CoV-2 BA.2.86 variant RBD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6779)
  AUTHORS   Taylor AL, Starr TN
  TITLE     Deep mutational scanning of SARS-CoV-2 Omicron BA.2.86 and epistatic
            emergence of the KP.3 variant.
  JOURNAL   Virus Evol. 2024 Sep 2;10(1):veae067. doi: 10.1093/ve/veae067. 
            eCollection 2024.
  PUBMED    39310091
REFERENCE   2  (bases 1 to 6779)
  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: "Virus 
            Evol."; date: "2024-09-2"; volume: "10(1):veae067. doi"; pages: " 
            10.1093/ve/veae067. eCollection 2024"
COMMENT     Sequence Label: 297_pETcon_SARS2_Omicron-BA286
FEATURES             Location/Qualifiers
     source          1..6779
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    2..18
                     /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)."
     primer_bind     7..29
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     26..42
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     26..42
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        63..81
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        111..763
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    315..432
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     727..750
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1082..1093
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1094..1120
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1808..1837
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2166..2185)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2167..2185)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      2383..2838
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2470..2489)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2680..2701
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2941..3615)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(3616..3896)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3917..3936)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4036..4058
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4096..4114)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4155..4658
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4695..4799
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4800..4868,4869..5660)
                     /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     complement(5018..5037)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5831..6419
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6320..6339
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6573..6590
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6707..6728
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6743..6748,6749..6766,6767..6773)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
       61 gaaattaacc ctcactaaag ggaacaaaag ctggtaccaa ttccttgaat tttcaaaaat
      121 tcttactttt tttttggatg gacgcaaaga agtttaataa tcatattaca tggcattacc
      181 accatataca tatccatatc taatcttact tatatgttgt ggaaatgtaa agagccccat
      241 tatcttagcc taaaaaaacc ttctctttgg aactttcagt aatacgctta actgctcatt
      301 gctatattga agtacggatt agaagccgcc gagcgggtga cagccctccg aaggaagact
      361 ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg tgcctcgcgc
      421 cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat gaagaggaaa
      481 aattggcagt aacctggccc cacaaacctt caaatgaacg aatcaaatta acaaccatag
      541 gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag cgaagcgatg
      601 atttttgatc tattaacaga tatataaatg caaaaactgc ataaccactt taactaatac
      661 tttcaacatt ttcggtttgt attacttctt attcaaatgt aataaaagta tcaacaaaaa
      721 attgttaata tacctctata ctttaacgtc aaggagaaaa aaccccggat cgaattccct
      781 acttcataca ttttcaatta agatgcagtt acttcgctgt ttttcaatat tttctgttat
      841 tgcttcagtt ttagcacagg aactgacaac tatatgcgag caaatcccct caccaacttt
      901 agaatcgacg ccgtactctt tgtcaacgac tactattttg gccaacggga aggcaatgca
      961 aggagttttt gaatattaca aatcagtaac gtttgtcagt aattgcggtt ctcacccctc
     1021 aacaactagc aaaggcagcc ccataaacac acagtatgtt tttaaggaca atagctcgac
     1081 gattgaaggt agatacccat acgacgttcc agactacgct ctgcaggcta gtggtggagg
     1141 aggctctggt ggaggcggcc gcggaggcgg agggtcggct agccatatga acgttaccaa
     1201 cttgtgtcca ttccatgaag ttttcaatgc tactagattc gcttctgttt acgcttggaa
     1261 tagaactaga atctctaact gcgttgctga ctattctgtc ttgtacaatt ttgctccatt
     1321 cttcgctttc aagtgctatg gtgtttctcc aactaagttg aacgatttgt gtttcaccaa
     1381 cgtttacgcc gattcctttg ttattaaagg taacgaagtc tcccaaattg ctccaggtca
     1441 aactggtaat attgccgatt acaattacaa gttgccagat gatttcaccg gttgtgttat
     1501 tgcttggaac tctaacaagt tggattctaa gcattctggc aactacgatt actggtacag
     1561 gttgttccgt aagtccaaat tgaagccatt cgaaagagat atttccaccg aaatctatca
     1621 agctggtaac aagccatgta aaggtaaagg tccaaactgt tacttcccat tgcaatctta
     1681 cggtttcaga ccaacttatg gtgttggtca tcaaccatac agagttgttg ttttgtcttt
     1741 cgagttgttg catgctccag ctactgtttg tggtccaaag aaatctactc tcgagggggg
     1801 cggttccgaa caaaagctta tttctgaaga ggacttgtaa tagagatctg ataacaacag
     1861 tgtagatgta acaaaatcga ctttgttccc actgtacttt tagctcgtac aaaatacaat
     1921 atacttttca tttctccgta aacaacatgt tttcccatgt aatatccttt tctatttttc
     1981 gttccgttac caactttaca catactttat atagctattc acttctatac actaaaaaac
     2041 taagacaatt ttaattttgc tgcctgccat atttcaattt gttataaatt cctataattt
     2101 atcctattag tagctaaaaa aagatgaatg tgaatcgaat cctaagagaa ttgagctcca
     2161 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttagatcg gaagagcaca
     2221 cgtctgaact ccagtcactt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     2281 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     2341 gatcgccttt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg
     2401 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2461 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2521 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2581 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2641 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2701 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     2761 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     2821 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2881 acaccgcata gatcggcaag tgcacaaaca atacttaaat aaatactact cagtaataac
     2941 ctatttctta gcatttttga cgaaatttgc tattttgtta gagtctttta caccatttgt
     3001 ctccacacct ccgcttacat caacaccaat aacgccattt aatctaagcg catcaccaac
     3061 attttctggc gtcagtccac cagctaacat aaaatgtaag ctttcggggc tctcttgcct
     3121 tccaacccag tcagaaatcg agttccaatc caaaagttca cctgtcccac ctgcttctga
     3181 atcaaacaag ggaataaacg aatgaggttt ctgtgaagct gcactgagta gtatgttgca
     3241 gtcttttgga aatacgagtc ttttaataac tggcaaaccg aggaactctt ggtattcttg
     3301 ccacgactca tctccatgca gttggacgat atcaatgccg taatcattga ccagagccaa
     3361 aacatcctcc ttaggttgat tacgaaacac gccaaccaag tatttcggag tgcctgaact
     3421 atttttatat gcttttacaa gacttgaaat tttccttgca ataaccgggt caattgttct
     3481 ctttctattg ggcacacata taatacccag caagtcagca tcggaatcta gagcacattc
     3541 tgcggcctct gtgctctgca agccgcaaac tttcaccaat ggaccagaac tacctgtgaa
     3601 attaataaca gacatactcc aagctgcctt tgtgtgctta atcacgtata ctcacgtgct
     3661 caatagtcac caatgccctc cctcttggcc ctctcctttt cttttttcga ccgaattaat
     3721 tcttaatcgg caaaaaaaga aaagctccgg atcaagattg tacgtaaggt gacaagctat
     3781 ttttcaataa agaatatctt ccactactgc catctggcgt cataactgca aagtacacat
     3841 atattacgat gctgttctat taaatgcttc ctatattata tatatagtaa tgtcgtgatc
     3901 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     3961 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     4021 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     4081 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     4141 tggtttctta gacggatcgc ttgcctgtaa cttacacgcg cctcgtatct tttaatgatg
     4201 gaataatttg ggaatttact ctgtgtttat ttatttttat gttttgtatt tggattttag
     4261 aaagtaaata aagaaggtag aagagttacg gaatgaagaa aaaaaaataa acaaaggttt
     4321 aaaaaatttc aacaaaaagc gtactttaca tatatattta ttagacaaga aaagcagatt
     4381 aaatagatat acattcgatt aacgataagt aaaatgtaaa atcacaggat tttcgtgtgt
     4441 ggtcttctac acagacaaga tgaaacaatt cggcattaat acctgagagc aggaagagca
     4501 agataaaagg tagtatttgt tggcgatccc cctagagtct tttacatctt cggaaaacaa
     4561 aaactatttt ttctttaatt tcttttttta ctttctattt ttaatttata tatttatatt
     4621 aaaaaattta aattataatt atttttatag cacgtgatga aaaggaccca ggtggcactt
     4681 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4741 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4801 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4861 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4921 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4981 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5041 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5101 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5161 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5221 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5281 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5341 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5401 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5461 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5521 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5581 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5641 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5701 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5761 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5821 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5881 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5941 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6001 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6061 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6121 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6181 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6241 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6301 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6361 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6421 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6481 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     6541 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     6601 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     6661 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     6721 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtgga
//
