LOCUS       sequence-362479-        6776 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast surface display of the SARS-CoV-2 Omicron BA.1 variant RBD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6776)
  AUTHORS   Starr TN, Greaney AJ, Hannon WW, Loes AN, Hauser K, Dillen JR, Ferri
            E, Farrell AG, Dadonaite B, McCallum M, Matreyek KA, Corti D, 
            Veesler D, Snell G, Bloom JD
  TITLE     Shifting mutational constraints in the SARS-CoV-2 receptor-binding 
            domain during viral evolution.
  JOURNAL   Science. 2022 Jun 28:eabo7896. doi: 10.1126/science.abo7896.
  PUBMED    35762884
REFERENCE   2  (bases 1 to 6776)
  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: "Science. 
            2022 Jun 28:eabo7896. doi: 10.1126/science.abo7896."
COMMENT     Sequence Label: 3294_pETcon-SARS2-RBD_Omicron-BA1
FEATURES             Location/Qualifiers
     source          1..6776
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    27..43
                     /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     32..54
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     51..67
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     51..67
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        88..106
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        136..788
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    340..457
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     752..775
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1107..1118
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1119..1145
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1836..1865
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2194..2213)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2195..2213)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      2405..2860
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2492..2511)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2702..2723
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2963..3637)
                     /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(3638..3918)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3939..3958)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4058..4080
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4118..4136)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4177..4680
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4717..4821
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4822..4890,4891..5682)
                     /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(5040..5059)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5853..6441
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6342..6361
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6595..6612
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6729..6750
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6765..6770,6771..6776,1..12,13..19)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
       61 ctatgaccat gattacgcca agctcgaaat taaccctcac taaagggaac aaaagctggt
      121 accaattcct tgaattttca aaaattctta cttttttttt ggatggacgc aaagaagttt
      181 aataatcata ttacatggca ttaccaccat atacatatcc atatctaatc ttacttatat
      241 gttgtggaaa tgtaaagagc cccattatct tagcctaaaa aaaccttctc tttggaactt
      301 tcagtaatac gcttaactgc tcattgctat attgaagtac ggattagaag ccgccgagcg
      361 ggtgacagcc ctccgaagga agactctcct ccgtgcgtcc tcgtcttcac cggtcgcgtt
      421 cctgaaacgc agatgtgcct cgcgccgcac tgctccgaac aataaagatt ctacaatact
      481 agcttttatg gttatgaaga ggaaaaattg gcagtaacct ggccccacaa accttcaaat
      541 gaacgaatca aattaacaac cataggatga taatgcgatt agttttttag ccttatttct
      601 ggggtaatta atcagcgaag cgatgatttt tgatctatta acagatatat aaatgcaaaa
      661 actgcataac cactttaact aatactttca acattttcgg tttgtattac ttcttattca
      721 aatgtaataa aagtatcaac aaaaaattgt taatatacct ctatacttta acgtcaagga
      781 gaaaaaaccc cggatcgaat tccctacttc atacattttc aattaagatg cagttacttc
      841 gctgtttttc aatattttct gttattgctt cagttttagc acaggaactg acaactatat
      901 gcgagcaaat cccctcacca actttagaat cgacgccgta ctctttgtca acgactacta
      961 ttttggccaa cgggaaggca atgcaaggag tttttgaata ttacaaatca gtaacgtttg
     1021 tcagtaattg cggttctcac ccctcaacaa ctagcaaagg cagccccata aacacacagt
     1081 atgtttttaa ggacaatagc tcgacgattg aaggtagata cccatacgac gttccagact
     1141 acgctctgca ggctagtggt ggaggaggct ctggtggagg cggccgcgga ggcggagggt
     1201 cggctagcca tatgaacata acaaacttat gcccctttga cgaagtattt aatgctacta
     1261 gattcgcatc ggtttatgcc tggaatagaa agaggatcag taactgcgtt gctgattatt
     1321 ctgttttgta taacctggcc cctttcttca catttaagtg ctacggggtc tcgcctacca
     1381 aattaaacga tttatgcttc accaatgtgt acgccgattc ttttgtgatc aggggtgacg
     1441 aagttagaca gatcgctcca gggcaaactg gtaatattgc cgattacaac tacaagcttc
     1501 cagacgactt cactggttgc gtaatagcat ggaactcaaa caagttagac tcaaaggtct
     1561 caggaaatta taactacctg taccgtcttt tcaggaaatc aaatttgaag ccgttcgaaa
     1621 gggacatctc cacggagata tatcaggccg gtaacaagcc ctgcaatggc gtcgccggct
     1681 tcaactgtta cttcccccta aggtcatact ctttcaggcc tacatacgga gttggccatc
     1741 agccatacag agttgtggtt ttatctttcg agttgttgca cgcccctgct acggtttgtg
     1801 gtcctaagaa gtccactctc gaggggggcg gttccgaaca aaagcttatt tctgaagagg
     1861 acttgtaata gagatctgat aacaacagtg tagatgtaac aaaatcgact ttgttcccac
     1921 tgtactttta gctcgtacaa aatacaatat acttttcatt tctccgtaaa caacatgttt
     1981 tcccatgtaa tatccttttc tatttttcgt tccgttacca actttacaca tactttatat
     2041 agctattcac ttctatacac taaaaaacta agacaatttt aattttgctg cctgccatat
     2101 ttcaatttgt tataaattcc tataatttat cctattagta gctaaaaaaa gatgaatgtg
     2161 aatcgaatcc taagagaatt gagctccaat tcgccctata gtgagtcgta ttacaattca
     2221 ctggccgtcg ttagatcgga agagcacgaa ctccagtcac ttacaacgtc gtgactggga
     2281 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     2341 taatagcgaa gaggcccgca ccgatcgcct ttcccaacag ttgcgcagcc tgaatggcga
     2401 atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2461 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2521 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2581 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2641 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2701 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2761 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2821 tttaacgcga attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc
     2881 ttacgcatct gtgcggtatt tcacaccgca tagatcggca agtgcacaaa caatacttaa
     2941 ataaatacta ctcagtaata acctatttct tagcattttt gacgaaattt gctattttgt
     3001 tagagtcttt tacaccattt gtctccacac ctccgcttac atcaacacca ataacgccat
     3061 ttaatctaag cgcatcacca acattttctg gcgtcagtcc accagctaac ataaaatgta
     3121 agctttcggg gctctcttgc cttccaaccc agtcagaaat cgagttccaa tccaaaagtt
     3181 cacctgtccc acctgcttct gaatcaaaca agggaataaa cgaatgaggt ttctgtgaag
     3241 ctgcactgag tagtatgttg cagtcttttg gaaatacgag tcttttaata actggcaaac
     3301 cgaggaactc ttggtattct tgccacgact catctccatg cagttggacg atatcaatgc
     3361 cgtaatcatt gaccagagcc aaaacatcct ccttaggttg attacgaaac acgccaacca
     3421 agtatttcgg agtgcctgaa ctatttttat atgcttttac aagacttgaa attttccttg
     3481 caataaccgg gtcaattgtt ctctttctat tgggcacaca tataataccc agcaagtcag
     3541 catcggaatc tagagcacat tctgcggcct ctgtgctctg caagccgcaa actttcacca
     3601 atggaccaga actacctgtg aaattaataa cagacatact ccaagctgcc tttgtgtgct
     3661 taatcacgta tactcacgtg ctcaatagtc accaatgccc tccctcttgg ccctctcctt
     3721 ttcttttttc gaccgaatta attcttaatc ggcaaaaaaa gaaaagctcc ggatcaagat
     3781 tgtacgtaag gtgacaagct atttttcaat aaagaatatc ttccactact gccatctggc
     3841 gtcataactg caaagtacac atatattacg atgctgttct attaaatgct tcctatatta
     3901 tatatatagt aatgtcgtga tctatggtgc actctcagta caatctgctc tgatgccgca
     3961 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     4021 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4081 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     4141 taggttaatg tcatgataat aatggtttct tagacggatc gcttgcctgt aacttacacg
     4201 cgcctcgtat cttttaatga tggaataatt tgggaattta ctctgtgttt atttattttt
     4261 atgttttgta tttggatttt agaaagtaaa taaagaaggt agaagagtta cggaatgaag
     4321 aaaaaaaaat aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt
     4381 tattagacaa gaaaagcaga ttaaatagat atacattcga ttaacgataa gtaaaatgta
     4441 aaatcacagg attttcgtgt gtggtcttct acacagacaa gatgaaacaa ttcggcatta
     4501 atacctgaga gcaggaagag caagataaaa ggtagtattt gttggcgatc cccctagagt
     4561 cttttacatc ttcggaaaac aaaaactatt ttttctttaa tttctttttt tactttctat
     4621 ttttaattta tatatttata ttaaaaaatt taaattataa ttatttttat agcacgtgat
     4681 gaaaaggacc caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4741 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4801 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4861 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4921 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4981 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5041 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5101 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5161 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5221 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5281 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5341 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5401 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5461 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5521 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5581 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5641 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5701 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5761 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5821 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5881 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5941 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6001 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6061 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6121 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6181 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6241 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6301 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6361 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6421 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6481 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6541 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     6601 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     6661 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     6721 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttat
//
