LOCUS       sequence-364550-        5009 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression vector designed for expression, secretion, and display of
            proteins on the extracellular surface of Saccharomyces cerevisiae 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5009)
  AUTHORS   Kieke MC, Cho BK, Boder ET, Kranz DM, Wittrup KD
  TITLE     Isolation of anti-T cell receptor scFv mutants by yeast surface 
            display.
  JOURNAL   Protein Eng. 1997 Nov;10(11):1303-10.
  PUBMED    9514119
REFERENCE   2  (bases 1 to 5009)
  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: "Protein 
            Eng."; date: "1997-11"; volume: "10(11)"; pages: "1303-10"
COMMENT     Sequence Label: pYD1
FEATURES             Location/Qualifiers
     source          1..5009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     504..522
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        539..640
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             641..1315
                     /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"
     CDS             complement(1686..1703)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1713..1754)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(join(1833..1847,1848..1856))
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             complement(1860..1889)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="ASMTGGQQMG"
     primer_bind     complement(2255..2274)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2256..2274)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2306..2747)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(2312..2335)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    2630..2747
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     promoter        complement(2766..2784)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2805..2821)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2805..2821)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2818..2840)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2829..2845
                     /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)."
     promoter        complement(join(2853..2859,2860..2877,2878..2883))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2898..2919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3036..3053)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3207..3795)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3287..3306)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3966..4757,4758..4826))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4589..4608
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4827..4931)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    join(4968..5009,1..462)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 taaatatata aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt
       61 tttccgaaga tgtaaaagac tctaggggga tcgccaacaa atactacctt ttatcttgct
      121 cttcctgctc tcaggtatta atgccgaatt gtttcatctt gtctgtgtag aagaccacac
      181 acgaaaatcc tgtgatttta cattttactt atcgttaatc gaatgtatat ctatttaatc
      241 tgcttttctt gtctaataaa tatatatgta aagtacgctt tttgttgaaa ttttttaaac
      301 ctttgtttat ttttttttct tcattccgta actcttctac cttctttatt tactttctaa
      361 aatccaaata caaaacataa aaataaataa acacagagta aattcccaaa ttattccatc
      421 attaaaagat acgaggcgcg tgtaagttac aggcaagcga tccgtcctaa gaaaccatta
      481 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcaagaaa
      541 ttcggtcgaa aaaagaaaag gagagggcca agagggaggg cattggtgac tattgagcac
      601 gtgagtatac gtgattaagc acacaaaggc agcttggagt atgtctgtta ttaatttcac
      661 aggtagttct ggtccattgg tgaaagtttg cggcttgcag agcacagagg ccgcagaatg
      721 tgctctagat tccgatgctg acttgctggg tattatatgt gtgcccaata gaaagagaac
      781 aattgacccg gttattgcaa ggaaaatttc aagtcttgta aaagcatata aaaatagttc
      841 aggcactccg aaatacttgg ttggcgtgtt tcgtaatcaa cctaaggagg atgttttggc
      901 tctggtcaat gattacggca ttgatatcgt ccaactgcac ggagatgagt cgtggcaaga
      961 ataccaagag ttcctcggtt tgccagttat taaaagactc gtatttccaa aagactgcaa
     1021 catactactc agtgcagctt cacagaaacc tcattcgttt attcccttgt ttgattcaga
     1081 agcaggtggg acaggtgaac ttttggattg gaactcgatt tctgactggg ttggaaggca
     1141 agagagcccc gagagcttac attttatgtt agctggtgga ctgacgccag aaaatgttgg
     1201 tgatgcgctt agattaaatg gcgttattgg tgttgatgta agcggaggtg tggagacaaa
     1261 tggtgtaaaa gactctaaca aaatagcaaa tttcgtcaaa aatgctaaga aataggttat
     1321 tactgagtag tatttattta agtattgttt gtgcacttgc caattctctt aggattcgat
     1381 tcacattcat ctttttttag ctactaatag gataaattat aggaatttat aacaaattga
     1441 aatatggcag gcagcaaaat taaaattgtc ttagtttttt agtgtataga agtgaatagc
     1501 tatataaagt atgtgtaaag ttggtaacgg aacgaaaaat agaaaaggat attacatggg
     1561 aaaacatgtt gtttacggag aaatgaaaag tatattgtat tttgtacgag ctaaaagtac
     1621 agtgggaaca aagtcgattt tgttacatct acactgttgt tatcagatca gcgggtttaa
     1681 actcaatggt gatggtgatg atgaccggta cgcgtagaat cgagaccgag gagagggtta
     1741 gggataggct taccttcgaa gggccctcta gactcgagcg gccgccactg tgctggatat
     1801 ctgcagaatt ccaccacact ggatcctggt accttatcgt catcgtcgta cagatcccga
     1861 cccatttgct gtccaccagt catgctagca gaaccaccac caccagaacc accaccacca
     1921 gaaccaccac caccactagc ctgcagaagc ttaaaaacat actgtgtgtt tatggggctg
     1981 cctttgctag tcgttgaggg gtgagaaccg caattactga caaacgttac tgatttgtaa
     2041 tattcaaaaa ctccttgcat tgccttcccg ttggccaaaa tagtagtcgt tgacaaagag
     2101 tacggcgtcg attctaaagt tggtgagggg atttgctcgc atatagttgt cagttcctgt
     2161 gctaaaactg aagcaataac agaaaatatt gaaaaacagc gaagtaactg catcttaatt
     2221 gaaaatgtat gaagtaggga attagcttaa tattccctat agtgagtcgt attacagctg
     2281 ctagtagtcc gatccggggt tttttctcct tgacgttaaa gtatagaggt atattaacaa
     2341 ttttttgttg atacttttat tacatttgaa taagaagtaa tacaaaccga aaatgttgaa
     2401 agtattagtt aaagtggtta tgcagttttt gcatttatat atctgttaat agatcaaaaa
     2461 tcatcgcttc gctgattaat taccccagaa ataaggctaa aaaactaatc gcattatcat
     2521 cctatggttg ttaatttgat tcgttcattt gaaggtttgt ggggccaggt tactgccaat
     2581 ttttcctctt cataaccata aaagctagta ttgtagaatc tttattgttc ggagcagtgc
     2641 ggcgcgaggc acatctgcgt ttcaggaacg cgaccggtga agacgaggac gcacggagga
     2701 gagtcttcct tcggagggct gtcacccgct cggcggcttc taatccgtac tagcagcttt
     2761 tgttcccttt agtgagggtt aatttcgagc ttggcgtaat catggtcata gctgtttcct
     2821 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     2881 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     2941 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3001 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3061 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3121 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3181 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3241 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3301 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3361 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3421 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3481 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3541 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3601 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3661 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3721 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3781 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3841 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3901 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3961 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4021 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4081 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4141 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4201 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4261 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4321 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4381 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4441 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4501 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4561 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa
     4621 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4681 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4741 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4801 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4861 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4921 ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt ccttttcatc acgtgctata
     4981 aaaataatta taatttaaat tttttaata
//
