LOCUS       sequence-200254-        7849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TUP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7849)
  AUTHORS   Xia Z, Turner GC, Hwang CS, Byrd C, Varshavsky A
  TITLE     Amino acids induce peptide uptake via accelerated degradation of 
            CUP9, the transcriptional repressor of the PTR2 peptide transporter.
  JOURNAL   J Biol Chem. 2008 Oct 24;283(43):28958-68. doi: 
            10.1074/jbc.M803980200. Epub 2008 Aug 15.
  PUBMED    18708352
REFERENCE   2  (bases 1 to 7849)
  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: 
            "10.1074/jbc.M803980200"; journalName: "J Biol Chem"; date: 
            "2008-10-24- 24"; volume: "283"; issue: "43"; pages: "28958-68"
COMMENT     Sequence Label: pTUP1
FEATURES             Location/Qualifiers
     source          1..7849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        194..409
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             410..1213
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1344..1799)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1481..1502)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1693..1712
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1925..1947
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1939..1956
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1940..1956
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1966..1985
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1966..1984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2009..2025
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2010..2026
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5076..5096)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5076..5094)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5115..5131)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5115..5131)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5128..5150)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5139..5155
                     /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(5163..5169,5170..5187,5188..5193))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5208..5229
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5346..5363)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5517..6105)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5597..5616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6276..7067,7068..7136))
                     /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     6899..6918
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7137..7241)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7278..7781
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7822..7840
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc
      241 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
      301 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
      361 cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaatca tgtcgaaagc
      421 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
      481 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
      541 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
      601 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
      661 caatttttta ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca
      721 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
      781 ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc
      841 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata
      901 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc
      961 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
     1021 gggtttagat gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt
     1081 ctctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
     1141 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
     1201 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
     1261 agcttcaatt taattatatc agttattacc ctatgcggtg tgaaataccg cacagatgcg
     1321 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     1381 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     1441 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     1501 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     1561 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     1621 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     1681 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     1741 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     1801 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     1861 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     1921 ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta
     1981 tagggcgaat tgggtaccgg gccccccctc gaggtcgacg gtatcgatat tggagctaac
     2041 cctactgctt tagagaaggg aatcaaagat ctataaattt tatattcatg agcacatgta
     2101 gttttttttt ttttagtttt aaatataaga aaaaagatag tatgtatata tatatatata
     2161 catatacata taaaaggaaa gtaagcttat acaatacaca aataaattca tatcaaagaa
     2221 taatgaaccg caatattcag aaacacagga aaaggagggg aagggatgaa tggtgaggaa
     2281 agtaactgtt tgttttaaag cgtacctgga tcataacata acaaataagt agataaaaaa
     2341 gaatagagag aaagatataa tgacaatgaa ttgaagaata gtttagttag ttacatttgt
     2401 aaagtgttcc ttttgtgttc tgttcttaat ttggcgctat ttttttatac ttccaaatcc
     2461 ttgctttaca atcaccgcta ccagtagcaa aaacgttata ttctggaccc agcggagacc
     2521 cgtttgccac agccacagat ataactgaat tcctatgacc ttgcaacatc aataacggat
     2581 tgccggattt cttatcccaa aacaggacac cacgatcttt ggaaccggac aagatgtact
     2641 catcattttg tgtggtggcc acggacaata caaagtcttt atgcccgata tacgtaactt
     2701 cacaagtgcc ggaatttgga gttttcgaat cgctcttgtt gtttgcattc tgcaaattcc
     2761 agagcttaac agatctatct aatgagccgg atacaacgct ttgtccatct ctagtgaaga
     2821 caacgctata aacagagtcc ttgtggcctg taccggattc gttttccgaa tctagtcttt
     2881 ccaccaagaa tccggtctcg gaatcccaaa ctctcacagc acgatctaga gaaccagcag
     2941 cgatgtattt accatcacct ggtgatacag cgacggtggt aacaccatct tcaatggata
     3001 aagtcaatga acactggcct gtacgtaagt cccaaatacg aacggtacgg tcaccagaac
     3061 cggagactaa tttgtcacct gagggaaagt agtccaatga ataaatatct tgttcgtggc
     3121 cttgaagaat cataacaatc tttctatttt caatatccca aattctaatc agtctgtctt
     3181 cagcacctgt tgccaaaaat ttcccatctg gagaaaaaca cactgaacgg atatacaagt
     3241 cggatgatgg ggaagacgaa gtgttcaggt tttccacatc ttttgccaat tctgctgccg
     3301 aagtcatcgc tgtggtagta attgtggtgg tagtagtggt tgtcattgta ttgttatccg
     3361 tggacgtggt ggtgttattt tcagtgatcg aatttcgatg gttattggca gcagaatcgt
     3421 cagatagacg ggccaccaga gaaccatctg aaacgcgata cacttgagta gttttgttgc
     3481 agcctgtggc taagtattca ccatcgttac tgaacttcac gcaacaaaca actgaagtat
     3541 gatccaaaga tttgtgtaac tcaacgtcaa tttctcttgg tagtgccggg ttgtataaaa
     3601 tataataatc atttgtttgc ttcttcagag catcgggaac agactgggaa tctagatcca
     3661 aaaggaaagg tgggataggt ttagagtggt ttgctctttg attataaggg actaagtagt
     3721 gatcctggtg caaactagcc ggggtggcgt cttcctcctt aggtttgatt tcagtttcag
     3781 ttgcggtggt agtggtggcc gtggtggcgg atccggtgtc atttatcttc gaggtattat
     3841 tattttccgg ttcagtttct ttcaacgtag attcaggtgc cttgacagag ggtaaagtag
     3901 tggtctcttt agaagtagga gatccgttga tggctgtgtt actcaatggt gccacggaaa
     3961 cctggggagg tggttgctgt tgttgaagtt gctgctgttg cagttgctgt tgtggtagtt
     4021 gttgttgggc gtttgaggac accacaggca aagtggtggt aggcaactgt gagccaacca
     4081 gattaggacg gctagcttgt actgggaagg ccgatggcga accagtagtt gtgtttgctg
     4141 ctggagtggc ggtggccgaa gtagtagccg gtggttgttg cgcaactgga acagatgcag
     4201 atgcagcggc tagctgctgc tgttgctgtt gtaaatgctg ctggacctgt tgctgctgtt
     4261 gctgttgccg ctgttgctgg acggtcaaag atgcaatttg atggtctctt tgctccagcc
     4321 ctagtttcaa gtgcttgatc tcttcttcgt acgcgtcctt cattttcctg tgagttagtt
     4381 ccagttcgta gacggtgttt cttatctgct gcatctcagc cagctgctgg ttcattttga
     4441 aatcgtaatc cttttggttt tgaagacggt aggtatttgc ctcttgtgag acttggagaa
     4501 actcctgtct gatggcatcg agaagctcat tcagcttatt ctgcgtattc gaaacgctgg
     4561 cagtcatatt ggtttggatg gaaagctgat ttctttcttc ccctgcttat caaaaaagac
     4621 aaaaaaacaa tagaaaaaga gtagtggtgt gcgaaaggta acttggcgtg aaagaaaatt
     4681 aaacaattgc agaaggacag atagaggggc acgtgttcag ccagttgttc ttgcttcgct
     4741 gatgtttatc aattgggggt agatggggga atttctctct atttgtttac ttatgtactt
     4801 gttattttgc tttgttgttt gttgctttga cgggagagct ttttaatttt ttcgcagttt
     4861 ttcagtgttt tttgaaaagt acgaaaaatt aaggtccgtc ggtagcgata atgtaagagg
     4921 gttctgtctc taaaaatgtt agggtgaaag ttccaaaaaa ttattaactc ctagattgct
     4981 cagccaaaaa gaagggtaaa ttagggcttt cctggtttat ttatttgact agttctagag
     5041 cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc
     5101 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5161 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5221 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5281 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5341 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5401 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5461 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5521 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5581 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5641 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5701 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5761 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5821 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5881 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5941 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6001 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6061 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6121 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6181 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6241 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6301 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6361 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6421 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6481 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6541 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6601 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6661 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6721 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6781 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6841 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6901 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6961 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7021 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7081 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7141 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7201 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7261 ccacctgggt ccttttcatc acgtgctata aaaataatta taatttaaat tttttaatat
     7321 aaatatataa attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt
     7381 ttccgaagat gtaaaagact ctagggggat cgccaacaaa tactaccttt tatcttgctc
     7441 ttcctgctct caggtattaa tgccgaattg tttcatcttg tctgtgtaga agaccacaca
     7501 cgaaaatcct gtgattttac attttactta tcgttaatcg aatgtatatc tatttaatct
     7561 gcttttcttg tctaataaat atatatgtaa agtacgcttt ttgttgaaat tttttaaacc
     7621 tttgtttatt tttttttctt cattccgtaa ctcttctacc ttctttattt actttctaaa
     7681 atccaaatac aaaacataaa aataaataaa cacagagtaa attcccaaat tattccatca
     7741 ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat ccgtctaaga aaccattatt
     7801 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc
//
