LOCUS       sequence-319556-        9041 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast display reporter for non-display yeast strains (TAG 
            construct).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9041)
  AUTHORS   Potts KA, Stieglitz JT, Lei M, Van Deventer JA
  TITLE     Reporter system architecture affects measurements of noncanonical 
            amino acid incorporation efficiency and fidelity.
  JOURNAL   Mol Syst Des Eng. 2020 Feb 1;5(2):573-588. doi: 10.1039/c9me00107g. 
            Epub 2020 Jan 23.
  PUBMED    33791108
REFERENCE   2  (bases 1 to 9041)
  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"; doi: 
            "10.1039/c9me00107g"; journalName: "Mol Syst Des Eng"; date: 
            "2020-02-1- 1"; volume: "5"; issue: "2"; pages: "573-588"
COMMENT     Sequence Label: pCTCON2-Aga1p-FAPB2.3.6L1TAG
FEATURES             Location/Qualifiers
     source          1..9041
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     CDS             768..779
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             780..806
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1608..1637
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1970..1989)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1971..1989)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1996..2013)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1996..2012)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2005..2027)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2153..2608
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2240..2259)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2450..2471
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2711..3385)
                     /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(3386..3666)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3687..3706)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3806..3828
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3866..3884)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3925..4428
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4465..4569
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4570..4638,4639..5430)
                     /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(4788..4807)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5601..6189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6090..6109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6343..6360
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6477..6498
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6513..6518,6519..6536,6537..6543)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6551..6567
                     /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     6556..6578
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6575..6591
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6575..6591
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6612..6630
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        join(8838..9041,1..449)
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
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 ccggatcgaa ttccctactt catacatttt
      481 caattaagat gcagttactt cgctgttttt caatattttc tgttattgct tcagttttag
      541 cacaggaact gacaactata tgcgagcaaa tcccctcacc aactttagaa tcgacgccgt
      601 actctttgtc aacgactact attttggcca acgggaaggc aatgcaagga gtttttgaat
      661 attacaaatc agtaacgttt gtcagtaatt gcggttctca cccctcaaca actagcaaag
      721 gcagccccat aaacacacag tatgttttta aggacaatag ctcgacgatt gaaggtagat
      781 acccatacga cgttccagac tacgctctgc aggctagtgg tggaggaggc tctggtggag
      841 gcggtagcgg aggcggaggg tcggctagct agatccagat gacccagtcc ccgagctccc
      901 tgtccgcctc tgtgagcgat agggtcacca tcacctgccg tgccggtcag tacggttctt
      961 cttacgtagc ctggtatcaa cagaaaccag gaaaagctcc gaagcttctg atttactctg
     1021 catcccacct ctactctgga gtcccttctc gcttctctgg tagccgttcc gggacgggtt
     1081 tcactctgac catcagcagt ctgcagccgg aagacttcgc aacttattac tgtcagcaac
     1141 cgggtccggc tccgggtctg atcacgttcg gacagggtac caaggtggag atcaaaggta
     1201 ccactgccgc tagtggtagt agtggtggca gtagcagtgg tgccgaggtt cggctggtgg
     1261 agtctggcgg tggcctggtg cagccagggg gctcactccg tttgtcctgt gcagcttctg
     1321 gcttcaacat ctactatggt tcttacatcc actgggtgcg tcaggccccg ggtaagggcc
     1381 tggaatgggt tgcaggtatt tctccttact acaactctac ttactatgcc gatagcgtca
     1441 agggccgttt cactataagc gcagacgcat ccaaaaacac agcctaccta caaatgaaca
     1501 gcttaagagc tgaggacact gccgtctatt attgtgctcg cggttacgct tctggtggta
     1561 tggactactg gggtcaagga accctggtca ccgtctcctc gggatccgaa caaaagctta
     1621 tttctgaaga ggacttgtaa tagctcgaga tctgataaca acagtgtaga tgtaacaaaa
     1681 tcgactttgt tcccactgta cttttagctc gtacaaaata caatatactt ttcatttctc
     1741 cgtaaacaac atgttttccc atgtaatatc cttttctatt tttcgttccg ttaccaactt
     1801 tacacatact ttatatagct attcacttct atacactaaa aaactaagac aattttaatt
     1861 ttgctgcctg ccatatttca atttgttata aattcctata atttatccta ttagtagcta
     1921 aaaaaagatg aatgtgaatc gaatcctaag agaattgagc tccaattcgc cctatagtga
     1981 gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     2041 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     2101 ggcccgcacc gatcgccttt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc
     2161 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2221 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2281 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2341 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2401 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2461 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2521 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2581 tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     2641 gcggtatttc acaccgcata gatcggcaag tgcacaaaca atacttaaat aaatactact
     2701 cagtaataac ctatttctta gcatttttga cgaaatttgc tattttgtta gagtctttta
     2761 caccatttgt ctccacacct ccgcttacat caacaccaat aacgccattt aatctaagcg
     2821 catcaccaac attttctggc gtcagtccac cagctaacat aaaatgtaag ctttcggggc
     2881 tctcttgcct tccaacccag tcagaaatcg agttccaatc caaaagttca cctgtcccac
     2941 ctgcttctga atcaaacaag ggaataaacg aatgaggttt ctgtgaagct gcactgagta
     3001 gtatgttgca gtcttttgga aatacgagtc ttttaataac tggcaaaccg aggaactctt
     3061 ggtattcttg ccacgactca tctccatgca gttggacgat atcaatgccg taatcattga
     3121 ccagagccaa aacatcctcc ttaggttgat tacgaaacac gccaaccaag tatttcggag
     3181 tgcctgaact atttttatat gcttttacaa gacttgaaat tttccttgca ataaccgggt
     3241 caattgttct ctttctattg ggcacacata taatacccag caagtcagca tcggaatcta
     3301 gagcacattc tgcggcctct gtgctctgca agccgcaaac tttcaccaat ggaccagaac
     3361 tacctgtgaa attaataaca gacatactcc aagctgcctt tgtgtgctta atcacgtata
     3421 ctcacgtgct caatagtcac caatgccctc cctcttggcc ctctcctttt cttttttcga
     3481 ccgaattaat tcttaatcgg caaaaaaaga aaagctccgg atcaagattg tacgtaaggt
     3541 gacaagctat ttttcaataa agaatatctt ccactactgc catctggcgt cataactgca
     3601 aagtacacat atattacgat gctgttctat taaatgcttc ctatattata tatatagtaa
     3661 tgtcgtgatc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     3721 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     3781 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     3841 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     3901 atgataataa tggtttctta gacggatcgc ttgcctgtaa cttacacgcg cctcgtatct
     3961 tttaatgatg gaataatttg ggaatttact ctgtgtttat ttatttttat gttttgtatt
     4021 tggattttag aaagtaaata aagaaggtag aagagttacg gaatgaagaa aaaaaaataa
     4081 acaaaggttt aaaaaatttc aacaaaaagc gtactttaca tatatattta ttagacaaga
     4141 aaagcagatt aaatagatat acattcgatt aacgataagt aaaatgtaaa atcacaggat
     4201 tttcgtgtgt ggtcttctac acagacaaga tgaaacaatt cggcattaat acctgagagc
     4261 aggaagagca agataaaagg tagtatttgt tggcgatccc cctagagtct tttacatctt
     4321 cggaaaacaa aaactatttt ttctttaatt tcttttttta ctttctattt ttaatttata
     4381 tatttatatt aaaaaattta aattataatt atttttatag cacgtgatga aaaggaccca
     4441 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4501 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4561 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4621 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4681 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4741 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4801 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4861 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4921 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4981 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5041 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5101 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5161 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5221 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5281 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5341 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5401 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5461 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5521 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5581 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5641 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5701 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5761 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5821 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5881 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5941 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6001 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6061 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6121 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6181 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6241 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6301 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6361 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6421 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     6481 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     6541 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     6601 gccaagctcg aaattaaccc tcactaaagg gaacaaaagc tgggtacctt aactgaaaat
     6661 tacattgcaa gcaactgcca tgatggcgaa gaccaacttt cctaatggca aacgcgtttg
     6721 cgatccactg cccatatatt ggctaattgt taccattgat gaagtttggt attgtatgga
     6781 gattgaagta gtggaaactt ctgtagtggt aacagatgtt tgtagagaag agggagcaga
     6841 tgatgatgag aggaaaatag aagataccgt cggagtcaat gatgaagatt caatagtcga
     6901 ttccacagag gtagatggtg tagagataga tgttgcggaa agtgatgatg tagaggcaga
     6961 tattgtagca atagctgtgg aagaagtggg ttgtaaaatc tctgatgttg aagaaatttg
     7021 aagtgtggta actggtgact gttcgacact tagtgatgat acagaactga atagcaaaat
     7081 agtactgtca gaggataatg tagcagacaa ggaggaggat gaaaggaccg atgtcgataa
     7141 taagtgtgat gattgttctt gagattctga gtcaatgctt gatgtactcg tggcttcact
     7201 tacactagtc gaaatggttg aggtacctga agatgttaca tgatacttac cggtagcgtg
     7261 cttattgaaa tcttcatccg aagttgtcat tgaacttacc caactctttg tggatgtggt
     7321 ctgctgtgtt acagaacttg atgaagctgt ttgcacggaa gaatacgagt gtgacgtttc
     7381 agcacatgtg gagcatgcat atgaggattc aaccgttgag gtgcttacag aagttgcgac
     7441 gtttttagtg gacattttta tagaagatgt aatcaaactt gttgtctgcg agggcatgca
     7501 agactcgtga gtgcagactg tagcgtctgt tgtaaccgtt tcatgcatac tggttaatgt
     7561 gctagtctca gattcagagc tataaggaca ccatgtcgta tacattgtag tgactccact
     7621 aacagtggtg aaatatgtag acatggaaaa tgatgggata gtagtagaaa ttgaggattt
     7681 ggtgatatat tcagacgaac tttgtgaact aacagttgtt tcaacagttg aagaagtcat
     7741 agaaggagtt gcaacatttg acgaagtcga agggacggag taaacaggtg tggatgggct
     7801 gtataatgac acagacgttg atgaagatgc tatagaggag ccaagggatg aagttgaatc
     7861 agtaaaagta gagctaatag atgttgaact tggagaagtg gaagccaaag ttggagatga
     7921 agaagtcaaa cttggggatg tagatgttga atatgaagaa gtggaagtgg agcttgcaga
     7981 agtagattta gaacttggag atgtagatgt tgaacttgat gaggtagatg tagagcttgg
     8041 agatgtagat gtcgaacttg atgaggtaga tgtagaactt tgggatgtag atgtagaact
     8101 tgaagatgtg gatgtcaaac ttgaagatgt ggatgtcgaa cttggagatg tagatgtcga
     8161 acttgatgag gtagatgtcg aacttgatga ggtagatgtc gaacttgatg aggtagatgt
     8221 agagcttgga gatgtagatg tagagcttgg agatgtagat gtcgaactta gggaggtagt
     8281 agttggattg gacgatgttg tactcgaaag tgtagaggtg acaggtgaga ttatagaggc
     8341 acttgatggt tcaatggcgc tggatgatac cacggttgta gttcctactt cggataatga
     8401 gctgatagcg gttgtgtgac aagtagggca aatatatgaa gtgaacttgg aagtcacaga
     8461 taaggtggtg gttggcaacg tcgatgacga aggacatgcc tcatgggagc agacttctgt
     8521 agataatgtc aaagtactaa atctggatag ggtagtagcg tacgatattg aaggacttat
     8581 ttcggcagca gatgaagtgg ataccgtcaa tggacaccaa gtcgtataca atgtcgtagt
     8641 gccagcttga acgatagtgg atgtggagac tagcgcgggt gaaactgtag ttgtaacaac
     8701 cccatttgca tcgtttgtct tggttatcgt cactagaatc gtttctggat cagatgccaa
     8761 ggcaatatta gttaatccca acaatattgt gaacaggtag gtaaaatgag cgaaagataa
     8821 tgtcatttat attgaatttt caaaaattct tacttttttt ttggatggac gcaaagaagt
     8881 ttaataatca tattacatgg cattaccacc atatacatat ccatatctaa tcttacttat
     8941 atgttgtgga aatgtaaaga gccccattat cttagcctaa aaaaaccttc tctttggaac
     9001 tttcagtaat acgcttaact gctcattgct atattgaagt a
//
