LOCUS       sequence-362502-        6771 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast surface display of the SARS-CoV-2 Omicron BA.2 variant RBD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6771)
  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 6771)
  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: 3322_pETcon_SARS2_Omicron-BA2
FEATURES             Location/Qualifiers
     source          1..6771
                     /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             1811..1840
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2169..2188)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2170..2188)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      2375..2830
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2462..2481)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2672..2693
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2933..3607)
                     /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(3608..3888)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3909..3928)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4028..4050
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4088..4106)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4147..4650
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4687..4791
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4792..4860,4861..5652)
                     /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(5010..5029)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5823..6411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6312..6331
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6565..6582
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6699..6720
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6735..6740,6741..6758,6759..6765)
                     /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 acatcaccaa
     1201 cttgtgtcca ttcgacgaag ttttcaatgc tactagattc gcttctgttt acgcttggaa
     1261 tagaaagaga atctctaact gcgttgctga ctattctgtc ttgtacaatt ttgctccatt
     1321 cttcgctttc aagtgctatg gtgtttctcc aactaagttg aacgatttgt gtttcaccaa
     1381 cgtttacgcc gattcctttg ttattagagg taacgaagtc tcccaaattg ctccaggtca
     1441 aactggtaat attgccgatt acaattacaa gttgccagat gatttcaccg gttgtgttat
     1501 tgcttggaac tctaacaagt tggattctaa ggttggtggc aactacaatt acttgtacag
     1561 gttgttccgt aagtccaact tgaagccatt cgaaagagat atttccaccg aaatctatca
     1621 agctggtaac aagccatgta atggtgttgc tggttttaac tgttacttcc cattgagatc
     1681 ttacggtttc agaccaactt atggtgttgg tcatcaacca tacagagttg ttgttttgtc
     1741 tttcgagttg ttgcatgctc cagctactgt ttgtggtcca aagaaatcta ctctcgaggg
     1801 gggcggttcc gaacaaaagc ttatttctga agaggacttg taatagagat ctgataacaa
     1861 cagtgtagat gtaacaaaat cgactttgtt cccactgtac ttttagctcg tacaaaatac
     1921 aatatacttt tcatttctcc gtaaacaaca tgttttccca tgtaatatcc ttttctattt
     1981 ttcgttccgt taccaacttt acacatactt tatatagcta ttcacttcta tacactaaaa
     2041 aactaagaca attttaattt tgctgcctgc catatttcaa tttgttataa attcctataa
     2101 tttatcctat tagtagctaa aaaaagatga atgtgaatcg aatcctaaga gaattgagct
     2161 ccaattcgcc ctatagtgag tcgtattaca attcactggc cgtcgttaga tccgtctgaa
     2221 ctccagtcac ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     2281 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgcct
     2341 ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg cgcattaagc
     2401 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2461 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2521 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2581 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2641 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2701 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2761 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     2821 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2881 tagatcggca agtgcacaaa caatacttaa ataaatacta ctcagtaata acctatttct
     2941 tagcattttt gacgaaattt gctattttgt tagagtcttt tacaccattt gtctccacac
     3001 ctccgcttac atcaacacca ataacgccat ttaatctaag cgcatcacca acattttctg
     3061 gcgtcagtcc accagctaac ataaaatgta agctttcggg gctctcttgc cttccaaccc
     3121 agtcagaaat cgagttccaa tccaaaagtt cacctgtccc acctgcttct gaatcaaaca
     3181 agggaataaa cgaatgaggt ttctgtgaag ctgcactgag tagtatgttg cagtcttttg
     3241 gaaatacgag tcttttaata actggcaaac cgaggaactc ttggtattct tgccacgact
     3301 catctccatg cagttggacg atatcaatgc cgtaatcatt gaccagagcc aaaacatcct
     3361 ccttaggttg attacgaaac acgccaacca agtatttcgg agtgcctgaa ctatttttat
     3421 atgcttttac aagacttgaa attttccttg caataaccgg gtcaattgtt ctctttctat
     3481 tgggcacaca tataataccc agcaagtcag catcggaatc tagagcacat tctgcggcct
     3541 ctgtgctctg caagccgcaa actttcacca atggaccaga actacctgtg aaattaataa
     3601 cagacatact ccaagctgcc tttgtgtgct taatcacgta tactcacgtg ctcaatagtc
     3661 accaatgccc tccctcttgg ccctctcctt ttcttttttc gaccgaatta attcttaatc
     3721 ggcaaaaaaa gaaaagctcc ggatcaagat tgtacgtaag gtgacaagct atttttcaat
     3781 aaagaatatc ttccactact gccatctggc gtcataactg caaagtacac atatattacg
     3841 atgctgttct attaaatgct tcctatatta tatatatagt aatgtcgtga tctatggtgc
     3901 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     3961 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4021 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     4081 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4141 tagacggatc gcttgcctgt aacttacacg cgcctcgtat cttttaatga tggaataatt
     4201 tgggaattta ctctgtgttt atttattttt atgttttgta tttggatttt agaaagtaaa
     4261 taaagaaggt agaagagtta cggaatgaag aaaaaaaaat aaacaaaggt ttaaaaaatt
     4321 tcaacaaaaa gcgtacttta catatatatt tattagacaa gaaaagcaga ttaaatagat
     4381 atacattcga ttaacgataa gtaaaatgta aaatcacagg attttcgtgt gtggtcttct
     4441 acacagacaa gatgaaacaa ttcggcatta atacctgaga gcaggaagag caagataaaa
     4501 ggtagtattt gttggcgatc cccctagagt cttttacatc ttcggaaaac aaaaactatt
     4561 ttttctttaa tttctttttt tactttctat ttttaattta tatatttata ttaaaaaatt
     4621 taaattataa ttatttttat agcacgtgat gaaaaggacc caggtggcac ttttcgggga
     4681 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4741 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4801 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4861 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4921 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4981 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5041 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5101 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5161 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5221 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5281 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5341 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5401 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5461 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5521 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5581 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5641 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5701 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5761 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5821 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5881 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5941 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6001 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6061 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6121 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6181 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6241 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6301 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6361 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6421 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     6481 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     6541 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     6601 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     6661 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     6721 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg a
//
