LOCUS       sequence-341486-        6083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast surface display plasmid backbone that can be combined with 
            pJS699 through homologous recombination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6083)
  AUTHORS   Francino-Urdaniz IM, Steiner PJ, Kirby MB, Zhao F, Haas CM, Barman 
            S, Rhodes ER, Leonard AC, Peng L, Sprenger KG, Jardine JG, Whitehead
            TA
  TITLE     One-shot identification of SARS-CoV-2 S RBD escape mutants using 
            yeast screening.
  JOURNAL   Cell Rep. 2021 Aug 31;36(9):109627. doi: 
            10.1016/j.celrep.2021.109627. Epub 2021 Aug 10.
  PUBMED    34416153
REFERENCE   2  (bases 1 to 6083)
  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: "Cell Rep.";
            date: "2021-08-31"; volume: "36(9):109627. doi"; pages: " 
            10.1016/j.celrep.2021.109627. Epub 2021 Aug 10"
COMMENT     Sequence Label: pJS697
FEATURES             Location/Qualifiers
     source          1..6083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        136..154
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        184..836
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    388..505
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     800..823
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1155..1166
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1167..1193
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1256..1285
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1614..1633)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1615..1633)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1640..1657)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1640..1656)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1649..1671)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1797..2252
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1884..1903)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2094..2115
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2355..3029)
                     /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(3030..3310)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3331..3350)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3450..3472
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3510..3528)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3569..4072
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4109..4213
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4214..5029
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4214..5029
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4287..4306)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5208..5796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5697..5716
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5950..5967
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaag ctcgaaatta accctcacta aagggaacaa aagctggtac caattccttg
      181 aattttcaaa aattcttact ttttttttgg atggacgcaa agaagtttaa taatcatatt
      241 acatggcatt accaccatat acatatccat atctaatctt acttatatgt tgtggaaatg
      301 taaagagccc cattatctta gcctaaaaaa accttctctt tggaactttc agtaatacgc
      361 ttaactgctc attgctatat tgaagtacgg attagaagcc gccgagcggg tgacagccct
      421 ccgaaggaag actctcctcc gtgcgtcctc gtcttcaccg gtcgcgttcc tgaaacgcag
      481 atgtgcctcg cgccgcactg ctccgaacaa taaagattct acaatactag cttttatggt
      541 tatgaagagg aaaaattggc agtaacctgg ccccacaaac cttcaaatga acgaatcaaa
      601 ttaacaacca taggatgata atgcgattag ttttttagcc ttatttctgg ggtaattaat
      661 cagcgaagcg atgatttttg atctattaac agatatataa atgcaaaaac tgcataacca
      721 ctttaactaa tactttcaac attttcggtt tgtattactt cttattcaaa tgtaataaaa
      781 gtatcaacaa aaaattgtta atatacctct atactttaac gtcaaggaga aaaaaccccg
      841 gatcgaattc cctacttcat acattttcaa ttaagatgca gttacttcgc tgtttttcaa
      901 tattttctgt tattgcttca gttttagcac aggaactgac aactatatgc gagcaaatcc
      961 cctcaccaac tttagaatcg acgccgtact ctttgtcaac gactactatt ttggccaacg
     1021 ggaaggcaat gcaaggagtt tttgaatatt acaaatcagt aacgtttgtc agtaattgcg
     1081 gttctcaccc ctcaacaact agcaaaggca gccccataaa cacacagtat gtttttaagg
     1141 acaatagctc gacgattgaa ggtagatacc catacgacgt tccagactac gctctgcagg
     1201 ctagtagaga ccatcactga acgttgttat ggagcgggtc tcagggggcg gatccgaaca
     1261 aaagcttatt tctgaagagg acttgtaata gagatctgat aacaacagtg tagatgtaac
     1321 aaaatcgact ttgttcccac tgtactttta gctcgtacaa aatacaatat acttttcatt
     1381 tctccgtaaa caacatgttt tcccatgtaa tatccttttc tatttttcgt tccgttacca
     1441 actttacaca tactttatat agctattcac ttctatacac taaaaaacta agacaatttt
     1501 aattttgctg cctgccatat ttcaatttgt tataaattcc tataatttat cctattagta
     1561 gctaaaaaaa gatgaatgtg aatcgaatcc taagagaatt gagctccaat tcgccctata
     1621 gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     1681 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     1741 aagaggcccg caccgatcgc ctttcccaac agttgcgcag cctgaatggc gaatggacgc
     1801 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1861 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     1921 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     1981 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2041 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2101 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2161 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2221 gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat
     2281 ctgtgcggta tttcacaccg catagatcgg caagtgcaca aacaatactt aaataaatac
     2341 tactcagtaa taacctattt cttagcattt ttgacgaaat ttgctatttt gttagagtct
     2401 tttacaccat ttgtctccac acctccgctt acatcaacac caataacgcc atttaatcta
     2461 agcgcatcac caacattttc tggcgtcagt ccaccagcta acataaaatg taagctttcg
     2521 gggctctctt gccttccaac ccagtcagaa atcgagttcc aatccaaaag ttcacctgtc
     2581 ccacctgctt ctgaatcaaa caagggaata aacgaatgag gtttctgtga agctgcactg
     2641 agtagtatgt tgcagtcttt tggaaatacg agtcttttaa taactggcaa accgaggaac
     2701 tcttggtatt cttgccacga ctcatctcca tgcagttgga cgatatcaat gccgtaatca
     2761 ttgaccagag ccaaaacatc ctccttaggt tgattacgaa acacgccaac caagtatttc
     2821 ggagtgcctg aactattttt atatgctttt acaagacttg aaattttcct tgcaataacc
     2881 gggtcaattg ttctctttct attgggcaca catataatac ccagcaagtc agcatcggaa
     2941 tctagagcac attctgcggc ctctgtgctc tgcaagccgc aaactttcac caatggacca
     3001 gaactacctg tgaaattaat aacagacata ctccaagctg cctttgtgtg cttaatcacg
     3061 tatactcacg tgctcaatag tcaccaatgc cctccctctt ggccctctcc ttttcttttt
     3121 tcgaccgaat taattcttaa tcggcaaaaa aagaaaagct ccggatcaag attgtacgta
     3181 aggtgacaag ctatttttca ataaagaata tcttccacta ctgccatctg gcgtcataac
     3241 tgcaaagtac acatatatta cgatgctgtt ctattaaatg cttcctatat tatatatata
     3301 gtaatgtcgt gatctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3361 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3421 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3481 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     3541 tgtcatgata ataatggttt cttagacgga tcgcttgcct gtaacttaca cgcgcctcgt
     3601 atcttttaat gatggaataa tttgggaatt tactctgtgt ttatttattt ttatgttttg
     3661 tatttggatt ttagaaagta aataaagaag gtagaagagt tacggaatga agaaaaaaaa
     3721 ataaacaaag gtttaaaaaa tttcaacaaa aagcgtactt tacatatata tttattagac
     3781 aagaaaagca gattaaatag atatacattc gattaacgat aagtaaaatg taaaatcaca
     3841 ggattttcgt gtgtggtctt ctacacagac aagatgaaac aattcggcat taatacctga
     3901 gagcaggaag agcaagataa aaggtagtat ttgttggcga tccccctaga gtcttttaca
     3961 tcttcggaaa acaaaaacta ttttttcttt aatttctttt tttactttct atttttaatt
     4021 tatatattta tattaaaaaa tttaaattat aattattttt atagcacgtg atgaaaagga
     4081 cccaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4141 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4201 aaaaaggaag agtatgagcc atattcaacg ggaaacgtct tgctctaggc cgcgattaaa
     4261 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     4321 aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     4381 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     4441 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     4501 actcaccact gcgatccccg ggaaaacagc attccaggta ttagaagaat atcctgattc
     4561 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     4621 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     4681 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     4741 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     4801 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     4861 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     4921 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     4981 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaac ctcagcaact
     5041 gtcaaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5101 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5161 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5221 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5281 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5341 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5401 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5461 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5521 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5581 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5641 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5701 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5761 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5821 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5881 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     5941 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     6001 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     6061 gcgggcagtg agcgcaacgc aat
//
