LOCUS       sequence-338367-        6090 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Displays MART-1 peptide in HLA-A*02 as a single-chain trimer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6090)
  AUTHORS   Huisman BD, Grace BE, Holec PV, Birnbaum ME
  TITLE     Yeast Display for the Identification of Peptide-MHC Ligands of 
            Immune Receptors.
  JOURNAL   Methods Mol Biol. 2022;2491:263-291. doi: 
            10.1007/978-1-0716-2285-8_15.
  PUBMED    35482196
REFERENCE   2  (bases 1 to 6090)
  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: "Methods Mol
            Biol."; date: "2022"; volume: "2491:263-291. doi"; pages: " 
            10.1007/978-1-0716-2285-8_15"
COMMENT     Sequence Label: pYAL_HLA-A2_MART1
FEATURES             Location/Qualifiers
     source          1..6090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..442
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1..118
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     413..436
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     474..493
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        474..492
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1387..1398
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1847..1876
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(2500..3174)
                     /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(3175..3276)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3293..3311)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3353..3857
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(3888..3908)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3888..3906)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3927..3943)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3927..3943)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3940..3962)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3951..3967
                     /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(3975..3981,3982..3999,4000..4005))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4020..4041
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4158..4175)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4329..4917)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4409..4428)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5088..5879,5880..5948))
                     /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     5711..5730
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5949..6053)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cggattagaa gccgccgagc gggtgacagc cctccgaagg aagactctcc tccgtgcgtc
       61 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa
      121 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc
      181 tggccccaca aaccttcaaa tgaacgaatc aaattaacaa ccataggatg ataatgcgat
      241 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt
      301 aacagatata taaatgcaaa aactgcataa ccactttaac taatactttc aacattttcg
      361 gtttgtatta cttcttattc aaatgtaata aaagtatcaa caaaaaattg ttaatatacc
      421 tctatacttt aacgtcaagg agaaaaaacc ccggatcgga ctactagcag ctgtaatacg
      481 actcactata gggaatatta agctaattct acttcataca ttttcaatta agatgcagtt
      541 acttcgctgt ttttcaatat tttctgttat tgctagcgtt ttggctgagg ccgcggggat
      601 aggtatattg acggtaggtg gaggaggttc tggaggtggt ggtagtggtg gtggtggttc
      661 catacaaaga actccaaaga tccaagttta cagtagacat cctgctgaaa acggtaaatc
      721 taatttcttg aactgttacg tctccggttt ccacccaagt gatatagaag ttgacttgtt
      781 gaaaaatggt gaaagaatcg aaaaggttga acattcagat ttgtcttttt ctaaggactg
      841 gtccttctat ttgttgtact acacagaatt cactccaact gaaaaggatg aatacgcttg
      901 cagagttaat catgtaacct tgtctcaacc taaaatcgtt aagtgggata gagacatggg
      961 tggtggtggt agtggtggtg gtggttcagg tggtggtggt tccggtggtg gtggttccca
     1021 tagtatgaga tatttcttta cttctgtttc aagaccaggt agaggtgaac ctagattcat
     1081 cgcagtcggt tacgttgatg acacacaatt tgtaagattc gattccgacg ctgcaagtca
     1141 aagaatggaa ccaagagcac cttggattga acaagaaggt ccagaatatt gggatggtga
     1201 aactagaaaa gttaaggccc attctcaaac tcacagagta gatttgggta cattaagagg
     1261 tgcttataat caatctgaag caggttcaca tacagtacaa agaatgtacg gttgtgatgt
     1321 cggttcagac tggagatttt tgagaggtta tcaccaatat gcttacgatg gtaaagacta
     1381 cattgcattg aaggaagatt tgagatcctg gaccgccgct gacatggcag cccaaactac
     1441 aaaacataag tgggaagctg cacacgtagc agaacaattg agagcctatt tggaaggtac
     1501 atgtgtcgaa tggttgagaa gatacttaga aaacggtaaa gaaacattgc aaagaaccga
     1561 tgctccaaag actcatatga cacatcacgc cgttagtgat cacgaagcta ctttgagatg
     1621 ctgggcatta tctttttacc ctgccgaaat cacattgacc tggcaaagag atggtgaaga
     1681 ccaaacccaa gatactgaat tagttgaaac cagaccagca ggtgacggta ctttccaaaa
     1741 atgggccgct gttgtagtcc cttcaggtca agaacaaaga tacacatgcc atgtccaaca
     1801 cgaaggttta ccaaagccat tgacattgag atgggaacca tcctctgaac aaaagcttat
     1861 ctccgaagaa gacttgggtg gtggtggatc tggtggtggt ggttctggtg gtggtggttc
     1921 tcaggaactg acaactatat gcgagcaaat cccctcacca actttagaat cgacgccgta
     1981 ctctttgtca acgactacta ttttggccaa cgggaaggca atgcaaggag tttttgaata
     2041 ttacaaatca gtaacgtttg tcagtaattg cggttctcac ccctcaacaa ctagcaaagg
     2101 cagccccata aacacacagt atgttttttg agtttaaacc cgctgatctg ataacaacag
     2161 tgtagatgta acaaaatcga ctttgttccc actgtacttt tagctcgtac aaaatacaat
     2221 atacttttca tttctccgta aacaacatgt tttcccatgt aatatccttt tctatttttc
     2281 gttccgttac caactttaca catactttat atagctattc acttctatac actaaaaaac
     2341 taagacaatt ttaattttgc tgcctgccat atttcaattt gttataaatt cctataattt
     2401 atcctattag tagctaaaaa aagatgaatg tgaatcgaat cctaagagaa ttgggcaagt
     2461 gcacaaacaa tacttaaata aatactactc agtaataacc tatttcttag catttttgac
     2521 gaaatttgct attttgttag agtcttttac accatttgtc tccacacctc cgcttacatc
     2581 aacaccaata acgccattta atctaagcgc atcaccaaca ttttctggcg tcagtccacc
     2641 agctaacata aaatgtaagc tctcggggct ctcttgcctt ccaacccagt cagaaatcga
     2701 gttccaatcc aaaagttcac ctgtcccacc tgcttctgaa tcaaacaagg gaataaacga
     2761 atgaggtttc tgtgaagctg cactgagtag tatgttgcag tcttttggaa atacgagtct
     2821 tttaataact ggcaaaccga ggaactcttg gtattcttgc cacgactcat ctccgtgcag
     2881 ttggacgata tcaatgccgt aatcattgac cagagccaaa acatcctcct taggttgatt
     2941 acgaaacacg ccaaccaagt atttcggagt gcctgaacta tttttatatg cttttacaag
     3001 acttgaaatt ttccttgcaa taaccgggtc aattgttctc tttctattgg gcacacatat
     3061 aatacccagc aagtcagcat cggaatctag agcacattct gcggcctctg tgctctgcaa
     3121 gccgcaaact ttcaccaatg gaccagaact acctgtgaaa ttaataacag acatactcca
     3181 agctgccttt gtgtgcttaa tcacgtatac tcacgtgctc aatagtcacc aatgccctcc
     3241 ctcttggccc tctccttttc ttttttcgac cgaatttctt gaagacgaaa gggcctcgtg
     3301 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagga cggatcgctt
     3361 gcctgtaact tacacgcgcc tcgtatcttt taatgatgga ataatttggg aatttactct
     3421 gtgtttattt atttttatgt tttgtatttg gattttagaa agtaaataaa gaaggtagaa
     3481 gagttacgga atgaagaaaa aaaaaataaa caaaggttta aaaaatttca acaaaaagcg
     3541 tactttacat atatatttat tagacaagaa aagcagatta aatagatata cattcgatta
     3601 acgataagta aaatgtaaaa tcacaggatt ttcgtgtgtg gtcttctaca cagacaagat
     3661 gaaacaattc ggcattaata cctgagagca ggaagagcaa gataaaaggt agtatttgtt
     3721 ggcgatcccc ctagagtctt ttacatcttc ggaaaacaaa aactattttt tctttaattt
     3781 ctttttttac tttctatttt taatttatat atttatatta aaaaatttaa attataatta
     3841 tttttatagc acgtgatgaa aaggaccccc taggccagct tttgttccct ttagtgaggg
     3901 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     3961 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4021 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4081 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4141 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4201 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4261 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4321 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4381 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4441 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4501 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4561 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4621 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4681 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4741 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4801 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4861 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4921 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4981 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5041 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5101 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5161 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5221 ataccgcgag aaccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5281 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5341 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5401 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5461 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5521 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5581 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5641 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5701 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5761 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5821 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5881 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5941 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6001 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6061 ccccgaaaag tgccacctac tagtctagta
//
