LOCUS       sequence-310730-        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"
     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        4982..5634
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    5186..5303
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     5598..5621
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             5953..5964
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             5965..5991
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6793..6822
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(7155..7174)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7156..7174)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7181..7198)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7181..7197)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7190..7212)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7338..7793
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7425..7444)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7635..7656
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7896..8570)
                     /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(8571..8851)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(8872..8891)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8991..9013
                     /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 aagctgggta ccttaactga aaattacatt gcaagcaact
     2821 gccatgatgg cgaagaccaa ctttcctaat ggcaaacgcg tttgcgatcc actgcccata
     2881 tattggctaa ttgttaccat tgatgaagtt tggtattgta tggagattga agtagtggaa
     2941 acttctgtag tggtaacaga tgtttgtaga gaagagggag cagatgatga tgagaggaaa
     3001 atagaagata ccgtcggagt caatgatgaa gattcaatag tcgattccac agaggtagat
     3061 ggtgtagaga tagatgttgc ggaaagtgat gatgtagagg cagatattgt agcaatagct
     3121 gtggaagaag tgggttgtaa aatctctgat gttgaagaaa tttgaagtgt ggtaactggt
     3181 gactgttcga cacttagtga tgatacagaa ctgaatagca aaatagtact gtcagaggat
     3241 aatgtagcag acaaggagga ggatgaaagg accgatgtcg ataataagtg tgatgattgt
     3301 tcttgagatt ctgagtcaat gcttgatgta ctcgtggctt cacttacact agtcgaaatg
     3361 gttgaggtac ctgaagatgt tacatgatac ttaccggtag cgtgcttatt gaaatcttca
     3421 tccgaagttg tcattgaact tacccaactc tttgtggatg tggtctgctg tgttacagaa
     3481 cttgatgaag ctgtttgcac ggaagaatac gagtgtgacg tttcagcaca tgtggagcat
     3541 gcatatgagg attcaaccgt tgaggtgctt acagaagttg cgacgttttt agtggacatt
     3601 tttatagaag atgtaatcaa acttgttgtc tgcgagggca tgcaagactc gtgagtgcag
     3661 actgtagcgt ctgttgtaac cgtttcatgc atactggtta atgtgctagt ctcagattca
     3721 gagctataag gacaccatgt cgtatacatt gtagtgactc cactaacagt ggtgaaatat
     3781 gtagacatgg aaaatgatgg gatagtagta gaaattgagg atttggtgat atattcagac
     3841 gaactttgtg aactaacagt tgtttcaaca gttgaagaag tcatagaagg agttgcaaca
     3901 tttgacgaag tcgaagggac ggagtaaaca ggtgtggatg ggctgtataa tgacacagac
     3961 gttgatgaag atgctataga ggagccaagg gatgaagttg aatcagtaaa agtagagcta
     4021 atagatgttg aacttggaga agtggaagcc aaagttggag atgaagaagt caaacttggg
     4081 gatgtagatg ttgaatatga agaagtggaa gtggagcttg cagaagtaga tttagaactt
     4141 ggagatgtag atgttgaact tgatgaggta gatgtagagc ttggagatgt agatgtcgaa
     4201 cttgatgagg tagatgtaga actttgggat gtagatgtag aacttgaaga tgtggatgtc
     4261 aaacttgaag atgtggatgt cgaacttgga gatgtagatg tcgaacttga tgaggtagat
     4321 gtcgaacttg atgaggtaga tgtcgaactt gatgaggtag atgtagagct tggagatgta
     4381 gatgtagagc ttggagatgt agatgtcgaa cttagggagg tagtagttgg attggacgat
     4441 gttgtactcg aaagtgtaga ggtgacaggt gagattatag aggcacttga tggttcaatg
     4501 gcgctggatg ataccacggt tgtagttcct acttcggata atgagctgat agcggttgtg
     4561 tgacaagtag ggcaaatata tgaagtgaac ttggaagtca cagataaggt ggtggttggc
     4621 aacgtcgatg acgaaggaca tgcctcatgg gagcagactt ctgtagataa tgtcaaagta
     4681 ctaaatctgg atagggtagt agcgtacgat attgaaggac ttatttcggc agcagatgaa
     4741 gtggataccg tcaatggaca ccaagtcgta tacaatgtcg tagtgccagc ttgaacgata
     4801 gtggatgtgg agactagcgc gggtgaaact gtagttgtaa caaccccatt tgcatcgttt
     4861 gtcttggtta tcgtcactag aatcgtttct ggatcagatg ccaaggcaat attagttaat
     4921 cccaacaata ttgtgaacag gtaggtaaaa tgagcgaaag ataatgtcat ttatattgaa
     4981 ttttcaaaaa ttcttacttt ttttttggat ggacgcaaag aagtttaata atcatattac
     5041 atggcattac caccatatac atatccatat ctaatcttac ttatatgttg tggaaatgta
     5101 aagagcccca ttatcttagc ctaaaaaaac cttctctttg gaactttcag taatacgctt
     5161 aactgctcat tgctatattg aagtacggat tagaagccgc cgagcgggtg acagccctcc
     5221 gaaggaagac tctcctccgt gcgtcctcgt cttcaccggt cgcgttcctg aaacgcagat
     5281 gtgcctcgcg ccgcactgct ccgaacaata aagattctac aatactagct tttatggtta
     5341 tgaagaggaa aaattggcag taacctggcc ccacaaacct tcaaatgaac gaatcaaatt
     5401 aacaaccata ggatgataat gcgattagtt ttttagcctt atttctgggg taattaatca
     5461 gcgaagcgat gatttttgat ctattaacag atatataaat gcaaaaactg cataaccact
     5521 ttaactaata ctttcaacat tttcggtttg tattacttct tattcaaatg taataaaagt
     5581 atcaacaaaa aattgttaat atacctctat actttaacgt caaggagaaa aaaccccgga
     5641 tcgaattccc tacttcatac attttcaatt aagatgcagt tacttcgctg tttttcaata
     5701 ttttctgtta ttgcttcagt tttagcacag gaactgacaa ctatatgcga gcaaatcccc
     5761 tcaccaactt tagaatcgac gccgtactct ttgtcaacga ctactatttt ggccaacggg
     5821 aaggcaatgc aaggagtttt tgaatattac aaatcagtaa cgtttgtcag taattgcggt
     5881 tctcacccct caacaactag caaaggcagc cccataaaca cacagtatgt ttttaaggac
     5941 aatagctcga cgattgaagg tagataccca tacgacgttc cagactacgc tctgcaggct
     6001 agtggtggag gaggctctgg tggaggcggt agcggaggcg gagggtcggc tagctagatc
     6061 cagatgaccc agtccccgag ctccctgtcc gcctctgtga gcgatagggt caccatcacc
     6121 tgccgtgccg gtcagtacgg ttcttcttac gtagcctggt atcaacagaa accaggaaaa
     6181 gctccgaagc ttctgattta ctctgcatcc cacctctact ctggagtccc ttctcgcttc
     6241 tctggtagcc gttccgggac gggtttcact ctgaccatca gcagtctgca gccggaagac
     6301 ttcgcaactt attactgtca gcaaccgggt ccggctccgg gtctgatcac gttcggacag
     6361 ggtaccaagg tggagatcaa aggtaccact gccgctagtg gtagtagtgg tggcagtagc
     6421 agtggtgccg aggttcggct ggtggagtct ggcggtggcc tggtgcagcc agggggctca
     6481 ctccgtttgt cctgtgcagc ttctggcttc aacatctact atggttctta catccactgg
     6541 gtgcgtcagg ccccgggtaa gggcctggaa tgggttgcag gtatttctcc ttactacaac
     6601 tctacttact atgccgatag cgtcaagggc cgtttcacta taagcgcaga cgcatccaaa
     6661 aacacagcct acctacaaat gaacagctta agagctgagg acactgccgt ctattattgt
     6721 gctcgcggtt acgcttctgg tggtatggac tactggggtc aaggaaccct ggtcaccgtc
     6781 tcctcgggat ccgaacaaaa gcttatttct gaagaggact tgtaatagct cgagatctga
     6841 taacaacagt gtagatgtaa caaaatcgac tttgttccca ctgtactttt agctcgtaca
     6901 aaatacaata tacttttcat ttctccgtaa acaacatgtt ttcccatgta atatcctttt
     6961 ctatttttcg ttccgttacc aactttacac atactttata tagctattca cttctataca
     7021 ctaaaaaact aagacaattt taattttgct gcctgccata tttcaatttg ttataaattc
     7081 ctataattta tcctattagt agctaaaaaa agatgaatgt gaatcgaatc ctaagagaat
     7141 tgagctccaa ttcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac
     7201 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     7261 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cctttcccaa cagttgcgca
     7321 gcctgaatgg cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     7381 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     7441 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     7501 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     7561 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     7621 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     7681 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     7741 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg
     7801 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagatcg gcaagtgcac
     7861 aaacaatact taaataaata ctactcagta ataacctatt tcttagcatt tttgacgaaa
     7921 tttgctattt tgttagagtc ttttacacca tttgtctcca cacctccgct tacatcaaca
     7981 ccaataacgc catttaatct aagcgcatca ccaacatttt ctggcgtcag tccaccagct
     8041 aacataaaat gtaagctttc ggggctctct tgccttccaa cccagtcaga aatcgagttc
     8101 caatccaaaa gttcacctgt cccacctgct tctgaatcaa acaagggaat aaacgaatga
     8161 ggtttctgtg aagctgcact gagtagtatg ttgcagtctt ttggaaatac gagtctttta
     8221 ataactggca aaccgaggaa ctcttggtat tcttgccacg actcatctcc atgcagttgg
     8281 acgatatcaa tgccgtaatc attgaccaga gccaaaacat cctccttagg ttgattacga
     8341 aacacgccaa ccaagtattt cggagtgcct gaactatttt tatatgcttt tacaagactt
     8401 gaaattttcc ttgcaataac cgggtcaatt gttctctttc tattgggcac acatataata
     8461 cccagcaagt cagcatcgga atctagagca cattctgcgg cctctgtgct ctgcaagccg
     8521 caaactttca ccaatggacc agaactacct gtgaaattaa taacagacat actccaagct
     8581 gcctttgtgt gcttaatcac gtatactcac gtgctcaata gtcaccaatg ccctccctct
     8641 tggccctctc cttttctttt ttcgaccgaa ttaattctta atcggcaaaa aaagaaaagc
     8701 tccggatcaa gattgtacgt aaggtgacaa gctatttttc aataaagaat atcttccact
     8761 actgccatct ggcgtcataa ctgcaaagta cacatatatt acgatgctgt tctattaaat
     8821 gcttcctata ttatatatat agtaatgtcg tgatctatgg tgcactctca gtacaatctg
     8881 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     8941 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     9001 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg a
//
