LOCUS       sequence-319667-        7297 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression of Aga2p-TAPBPR-myc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7297)
  AUTHORS   McShan AC, Devlin CA, Morozov GI, Overall SA, Moschidi D, Akella N, 
            Procko E, Sgourakis NG
  TITLE     TAPBPR promotes antigen loading on MHC-I molecules using a peptide 
            trap.
  JOURNAL   Nat Commun. 2021 May 26;12(1):3174. doi: 10.1038/s41467-021-23225-6.
  PUBMED    34039964
REFERENCE   2  (bases 1 to 7297)
  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: "Nat 
            Commun."; date: "2021-05-26"; volume: "12(1):3174. doi"; pages: " 
            10.1038/s41467-021-23225-6"
COMMENT     Sequence Label: pETCON-Aga2p-TAPBPR-myc
FEATURES             Location/Qualifiers
     source          1..7297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    69..572
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        609..713
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(714..782,783..1574)
                     /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(932..951)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1745..2333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2234..2253
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2487..2504
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2621..2642
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2657..2662,2663..2680,2681..2687)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2695..2711
                     /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     2700..2722
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2719..2735
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2719..2735
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2756..2774
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2804..3456
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    3008..3125
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3420..3443
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             3775..3786
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3787..3813
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5053..5082
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(5411..5430)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5412..5430)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5437..5454)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5437..5453)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5446..5468)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5594..6049
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5681..5700)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5891..5912
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6152..6826)
                     /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(6827..7107)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(7128..7147)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7247..7269
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgga tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat gatggaataa
      121 tttgggaatt tactctgtgt ttatttattt ttatgttttg tatttggatt ttagaaagta
      181 aataaagaag gtagaagagt tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa
      241 tttcaacaaa aagcgtactt tacatatata tttattagac aagaaaagca gattaaatag
      301 atatacattc gattaacgat aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt
      361 ctacacagac aagatgaaac aattcggcat taatacctga gagcaggaag agcaagataa
      421 aaggtagtat ttgttggcga tccccctaga gtcttttaca tcttcggaaa acaaaaacta
      481 ttttttcttt aatttctttt tttactttct atttttaatt tatatattta tattaaaaaa
      541 tttaaattat aattattttt atagcacgtg atgaaaagga cccaggtggc acttttcggg
      601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      721 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      781 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      841 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      901 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
      961 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1021 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1081 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1141 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1201 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1261 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1321 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1381 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1441 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1501 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1561 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1621 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1681 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1741 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1801 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1861 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     1921 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1981 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2041 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2101 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2161 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2221 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2281 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2341 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2401 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2461 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2521 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     2581 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     2641 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     2701 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ctcgaaatta
     2761 accctcacta aagggaacaa aagctggtac caattccttg aattttcaaa aattcttact
     2821 ttttttttgg atggacgcaa agaagtttaa taatcatatt acatggcatt accaccatat
     2881 acatatccat atctaatctt acttatatgt tgtggaaatg taaagagccc cattatctta
     2941 gcctaaaaaa accttctctt tggaactttc agtaatacgc ttaactgctc attgctatat
     3001 tgaagtacgg attagaagcc gccgagcggg tgacagccct ccgaaggaag actctcctcc
     3061 gtgcgtcctc gtcttcaccg gtcgcgttcc tgaaacgcag atgtgcctcg cgccgcactg
     3121 ctccgaacaa taaagattct acaatactag cttttatggt tatgaagagg aaaaattggc
     3181 agtaacctgg ccccacaaac cttcaaatga acgaatcaaa ttaacaacca taggatgata
     3241 atgcgattag ttttttagcc ttatttctgg ggtaattaat cagcgaagcg atgatttttg
     3301 atctattaac agatatataa atgcaaaaac tgcataacca ctttaactaa tactttcaac
     3361 attttcggtt tgtattactt cttattcaaa tgtaataaaa gtatcaacaa aaaattgtta
     3421 atatacctct atactttaac gtcaaggaga aaaaaccccg gatcgaattc cctacttcat
     3481 acattttcaa ttaagatgca gttacttcgc tgtttttcaa tattttctgt tattgcttca
     3541 gttttagcac aggaactgac aactatatgc gagcaaatcc cctcaccaac tttagaatcg
     3601 acgccgtact ctttgtcaac gactactatt ttggccaacg ggaaggcaat gcaaggagtt
     3661 tttgaatatt acaaatcagt aacgtttgtc agtaattgcg gttctcaccc ctcaacaact
     3721 agcaaaggca gccccataaa cacacagtat gtttttaagg acaatagctc gacgattgaa
     3781 ggtagatacc catacgacgt tccagactac gctctgcagg ctagtggtgg aggaggctct
     3841 ggtggaggcg gtagcggagg cggagggtcg gctagccata tgaagcccca cccagcagag
     3901 gggcagtggc gggcagtgga cgtggtccta gactgtttcc tggtgaagga cggtgcgcac
     3961 cgtggagctc tcgccagcag tgaggacagg gcaagggcct cccttgtgct gaagcaggtg
     4021 ccagtgctgg acgatggctc cctggaggac ttcaccgatt tccaaggggg cacactggcc
     4081 caagatgacc cacctattat ctttgaggcc tcagtggacc tggtccagat tccccaggcc
     4141 gaggccttgc tccatgctga ctgcagtggg aaggaggtga cctgtgagat ctcccgctac
     4201 tttctccaga tgacagagac cactgttaag acagcagctt ggttcatggc caacgtgcag
     4261 gtctctggag ggggacctag catctccttg gtgatgaaga ctcccagggt cgccaagaat
     4321 gaggtgctct ggcacccaac gctgaacttg ccactgagcc cccaggggac tgtgcgaact
     4381 gcagtggagt tccaggtgat gacacagacc caatccctga gcttcctgct ggggtcctca
     4441 gcctccttgg actgtggctt ctccatggca ccgggcttgg acctcatcag tgtggagtgg
     4501 cgactgcagc acaagggcag gggtcagttg gtgtacagct ggaccgcagg gcaggggcag
     4561 gctgtgcgga agggcgctac cctggagcct gcacaactgg gcatggccag ggatgcctcc
     4621 ctcaccctgc ccggcctcac tatacaggac gaggggacct acatttgcca gatcaccacc
     4681 tctctgtacc gagctcagca gatcatccag ctcaacatcc aagcttcccc taaagtacga
     4741 ctgagcttgg caaacgaagc tctgctgccc accctcatct gcgacattgc tggctattac
     4801 cctctggatg tggtggtgac gtggacccga gaggagctgg gtggatcccc agcccaagtc
     4861 tctggtgcct ccttctccag cctcaggcaa agcgtggcag gcacctacag catctcctcc
     4921 tctctcaccg cagaacctgg ctctgcaggt gccacttaca cctgccaggt cacacacatc
     4981 tctctggagg agccccttgg ggccagcacc caggttgtcc caccagagcg gagactcgag
     5041 gggggcggat ccgaacaaaa gcttatttct gaagaggact tgtaatagag atctgataac
     5101 aacagtgtag atgtaacaaa atcgactttg ttcccactgt acttttagct cgtacaaaat
     5161 acaatatact tttcatttct ccgtaaacaa catgttttcc catgtaatat ccttttctat
     5221 ttttcgttcc gttaccaact ttacacatac tttatatagc tattcacttc tatacactaa
     5281 aaaactaaga caattttaat tttgctgcct gccatatttc aatttgttat aaattcctat
     5341 aatttatcct attagtagct aaaaaaagat gaatgtgaat cgaatcctaa gagaattgag
     5401 ctccaattcg ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg
     5461 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     5521 cagctggcgt aatagcgaag aggcccgcac cgatcgcctt tcccaacagt tgcgcagcct
     5581 gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5641 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     5701 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     5761 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     5821 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     5881 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     5941 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     6001 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt
     6061 attttctcct tacgcatctg tgcggtattt cacaccgcat agatcggcaa gtgcacaaac
     6121 aatacttaaa taaatactac tcagtaataa cctatttctt agcatttttg acgaaatttg
     6181 ctattttgtt agagtctttt acaccatttg tctccacacc tccgcttaca tcaacaccaa
     6241 taacgccatt taatctaagc gcatcaccaa cattttctgg cgtcagtcca ccagctaaca
     6301 taaaatgtaa gctttcgggg ctctcttgcc ttccaaccca gtcagaaatc gagttccaat
     6361 ccaaaagttc acctgtccca cctgcttctg aatcaaacaa gggaataaac gaatgaggtt
     6421 tctgtgaagc tgcactgagt agtatgttgc agtcttttgg aaatacgagt cttttaataa
     6481 ctggcaaacc gaggaactct tggtattctt gccacgactc atctccatgc agttggacga
     6541 tatcaatgcc gtaatcattg accagagcca aaacatcctc cttaggttga ttacgaaaca
     6601 cgccaaccaa gtatttcgga gtgcctgaac tatttttata tgcttttaca agacttgaaa
     6661 ttttccttgc aataaccggg tcaattgttc tctttctatt gggcacacat ataataccca
     6721 gcaagtcagc atcggaatct agagcacatt ctgcggcctc tgtgctctgc aagccgcaaa
     6781 ctttcaccaa tggaccagaa ctacctgtga aattaataac agacatactc caagctgcct
     6841 ttgtgtgctt aatcacgtat actcacgtgc tcaatagtca ccaatgccct ccctcttggc
     6901 cctctccttt tcttttttcg accgaattaa ttcttaatcg gcaaaaaaag aaaagctccg
     6961 gatcaagatt gtacgtaagg tgacaagcta tttttcaata aagaatatct tccactactg
     7021 ccatctggcg tcataactgc aaagtacaca tatattacga tgctgttcta ttaaatgctt
     7081 cctatattat atatatagta atgtcgtgat ctatggtgca ctctcagtac aatctgctct
     7141 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     7201 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     7261 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcga
//
