LOCUS       sequence-327026-        6084 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 6084)
  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 6084)
  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..6084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..24,25..31)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    39..55
                     /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     44..66
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     63..79
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        100..118
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        148..800
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    352..469
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     764..787
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1119..1130
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1131..1157
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1220..1249
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1578..1597)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1579..1597)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1604..1621)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1604..1620)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1613..1635)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1761..2216
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1848..1867)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2058..2079
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2319..2993)
                     /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(2994..3274)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3295..3314)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3414..3436
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3474..3492)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3533..4036
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4073..4177
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4178..4993
                     /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             4178..4993
                     /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(4251..4270)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5173..5761
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5662..5681
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5915..5932
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6049..6070
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
       61 cacaggaaac agctatgacc atgattacgc caagctcgaa attaaccctc actaaaggga
      121 acaaaagctg gtaccaattc cttgaatttt caaaaattct tacttttttt ttggatggac
      181 gcaaagaagt ttaataatca tattacatgg cattaccacc atatacatat ccatatctaa
      241 tcttacttat atgttgtgga aatgtaaaga gccccattat cttagcctaa aaaaaccttc
      301 tctttggaac tttcagtaat acgcttaact gctcattgct atattgaagt acggattaga
      361 agccgccgag cgggtgacag ccctccgaag gaagactctc ctccgtgcgt cctcgtcttc
      421 accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga acaataaaga
      481 ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac ctggccccac
      541 aaaccttcaa atgaacgaat caaattaaca accataggat gataatgcga ttagtttttt
      601 agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat taacagatat
      661 ataaatgcaa aaactgcata accactttaa ctaatacttt caacattttc ggtttgtatt
      721 acttcttatt caaatgtaat aaaagtatca acaaaaaatt gttaatatac ctctatactt
      781 taacgtcaag gagaaaaaac cccggatcga attccctact tcatacattt tcaattaaga
      841 tgcagttact tcgctgtttt tcaatatttt ctgttattgc ttcagtttta gcacaggaac
      901 tgacaactat atgcgagcaa atcccctcac caactttaga atcgacgccg tactctttgt
      961 caacgactac tattttggcc aacgggaagg caatgcaagg agtttttgaa tattacaaat
     1021 cagtaacgtt tgtcagtaat tgcggttctc acccctcaac aactagcaaa ggcagcccca
     1081 taaacacaca gtatgttttt aaggacaata gctcgacgat tgaaggtaga tacccatacg
     1141 acgttccaga ctacgctctg caggctagta gagaccatca ctgaacgttg ttatggagcg
     1201 ggtctcaggg ggcggatccg aacaaaagct tatttctgaa gaggacttgt aatagagatc
     1261 tgataacaac agtgtagatg taacaaaatc gactttgttc ccactgtact tttagctcgt
     1321 acaaaataca atatactttt catttctccg taaacaacat gttttcccat gtaatatcct
     1381 tttctatttt tcgttccgtt accaacttta cacatacttt atatagctat tcacttctat
     1441 acactaaaaa actaagacaa ttttaatttt gctgcctgcc atatttcaat ttgttataaa
     1501 ttcctataat ttatcctatt agtagctaaa aaaagatgaa tgtgaatcga atcctaagag
     1561 aattgagctc caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac
     1621 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     1681 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcctttcc caacagttgc
     1741 gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     1801 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     1861 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     1921 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     1981 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2041 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2101 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2161 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg
     2221 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataga tcggcaagtg
     2281 cacaaacaat acttaaataa atactactca gtaataacct atttcttagc atttttgacg
     2341 aaatttgcta ttttgttaga gtcttttaca ccatttgtct ccacacctcc gcttacatca
     2401 acaccaataa cgccatttaa tctaagcgca tcaccaacat tttctggcgt cagtccacca
     2461 gctaacataa aatgtaagct ttcggggctc tcttgccttc caacccagtc agaaatcgag
     2521 ttccaatcca aaagttcacc tgtcccacct gcttctgaat caaacaaggg aataaacgaa
     2581 tgaggtttct gtgaagctgc actgagtagt atgttgcagt cttttggaaa tacgagtctt
     2641 ttaataactg gcaaaccgag gaactcttgg tattcttgcc acgactcatc tccatgcagt
     2701 tggacgatat caatgccgta atcattgacc agagccaaaa catcctcctt aggttgatta
     2761 cgaaacacgc caaccaagta tttcggagtg cctgaactat ttttatatgc ttttacaaga
     2821 cttgaaattt tccttgcaat aaccgggtca attgttctct ttctattggg cacacatata
     2881 atacccagca agtcagcatc ggaatctaga gcacattctg cggcctctgt gctctgcaag
     2941 ccgcaaactt tcaccaatgg accagaacta cctgtgaaat taataacaga catactccaa
     3001 gctgcctttg tgtgcttaat cacgtatact cacgtgctca atagtcacca atgccctccc
     3061 tcttggccct ctccttttct tttttcgacc gaattaattc ttaatcggca aaaaaagaaa
     3121 agctccggat caagattgta cgtaaggtga caagctattt ttcaataaag aatatcttcc
     3181 actactgcca tctggcgtca taactgcaaa gtacacatat attacgatgc tgttctatta
     3241 aatgcttcct atattatata tatagtaatg tcgtgatcta tggtgcactc tcagtacaat
     3301 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     3361 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3421 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     3481 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cggatcgctt
     3541 gcctgtaact tacacgcgcc tcgtatcttt taatgatgga ataatttggg aatttactct
     3601 gtgtttattt atttttatgt tttgtatttg gattttagaa agtaaataaa gaaggtagaa
     3661 gagttacgga atgaagaaaa aaaaataaac aaaggtttaa aaaatttcaa caaaaagcgt
     3721 actttacata tatatttatt agacaagaaa agcagattaa atagatatac attcgattaa
     3781 cgataagtaa aatgtaaaat cacaggattt tcgtgtgtgg tcttctacac agacaagatg
     3841 aaacaattcg gcattaatac ctgagagcag gaagagcaag ataaaaggta gtatttgttg
     3901 gcgatccccc tagagtcttt tacatcttcg gaaaacaaaa actatttttt ctttaatttc
     3961 tttttttact ttctattttt aatttatata tttatattaa aaaatttaaa ttataattat
     4021 ttttatagca cgtgatgaaa aggacccagg tggcactttt cggggaaatg tgcgcggaac
     4081 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4141 ctgataaatg cttcaataat attgaaaaag gaagagtatg agccatattc aacgggaaac
     4201 gtcttgctct aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     4261 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga
     4321 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     4381 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     4441 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccgggaaaa cagcattcca
     4501 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     4561 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     4621 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     4681 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     4741 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     4801 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     4861 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     4921 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     4981 tgagtttttc taacctcagc aactgtcaga ccaagtttac tcatatatac tttagattga
     5041 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5101 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5161 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5221 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5281 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5341 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5401 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5461 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5521 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5581 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5641 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5701 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5761 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5821 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     5881 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     5941 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     6001 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     6061 gctcactcat taggcacccc aggc
//
