LOCUS       sequence-160273-        6822 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Destination vector for integration at yeast Trp locus with 
            N-terminal TPL repression domain (RD) for auxin response assays .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6822)
  AUTHORS   Pierre-Jerome E, Jang SS, Havens KA, Nemhauser JL, Klavins E
  TITLE     Recapitulation of the forward nuclear auxin response pathway in 
            yeast.
  JOURNAL   Proc Natl Acad Sci U S A. 2014 Jul 1;111(26):9407-12. doi: 
            10.1073/pnas.1324147111. Epub 2014 Jun 16.
  PUBMED    24979769
REFERENCE   2  (bases 1 to 6822)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1073/pnas.1324147111"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2014-07-1- 1"; volume: "111"; issue: "26"; pages: "9407-12"
COMMENT     Sequence Label: pGP4G-TPLRD1-ccdB
FEATURES             Location/Qualifiers
     source          1..6822
                     /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        296..443
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             444..1118
                     /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        1171..1814
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     1708..1731
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     1820..1836
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1820..1836
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    2173..2297
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2322..2327,2328..2345,2346..2352)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2406..3065
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2472..2491)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3410..3715
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3594..3613
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3756..3880)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3887..4133
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3903..3921)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4601..4617)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4601..4617)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4614..4636)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4625..4641
                     /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(4649..4655,4656..4673,4674..4679))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4832..4849)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5003..5591)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5762..6553,6554..6622))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6385..6404
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6623..6727)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6795..6813
                     /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 accagtttaa accatttaat agaacagcat cgtaatatat gtgtactttg cagttatgac
      241 gccagatggc agtagtggaa gatattcttt attgaaaaat agcttgtcac cttacgtaca
      301 atcttgatcc ggagcttttc tttttttgcc gattaagaat taattcggtc gaaaaaagaa
      361 aaggagaggg ccaagaggga gggcattggt gactattgag cacgtgagta tacgtgatta
      421 agcacacaaa ggcagcttgg agtatgtctg ttattaattt cacaggtagt tctggtccat
      481 tggtgaaagt ttgcggcttg cagagcacag aggccgcaga atgtgctcta gattccgatg
      541 ctgacttgct gggtattata tgtgtgccca atagaaagag aacaattgac ccggttattg
      601 caaggaaaat ttcaagtctt gtaaaagcat ataaaaatag ttcaggcact ccgaaatact
      661 tggttggcgt gtttcgtaat caacctaagg aggatgtttt ggctctggtc aatgattacg
      721 gcattgatat cgtccaactg catggagatg agtcgtggca agaataccaa gagttcctcg
      781 gtttgccagt tattaaaaga ctcgtatttc caaaagactg caacatacta ctcagtgcag
      841 cttcacagaa acctcattcg tttattccct tgtttgattc agaagcaggt gggacaggtg
      901 aacttttgga ttggaactcg atttctgact gggttggaag gcaagagagc cccgaaagct
      961 tacattttat gttagctggt ggactgacgc cagaaaatgt tggtgatgcg cttagattaa
     1021 atggcgttat tggtgttgat gtaagcggag gtgtggagac aaatggtgta aaagactcta
     1081 acaaaatagc aaatttcgtc aaaaatgcta agaaataggt tattactgag tagtatttat
     1141 ttaagtattg tttgtgcact tgccagttta tcattatcaa tactcgccat ttcaaagaat
     1201 acgtaaataa ttaatagtag tgattttcct aactttattt agtcaaaaaa ttagcctttt
     1261 aattctgctg taacccgtac atgcccaaaa tagggggcgg gttacacaga atatataaca
     1321 tcgtaggtgt ctgggtgaac agtttattcc tggcatccac taaatataat ggagcccgct
     1381 ttttaagctg gcatccagaa aaaaaaagaa tcccagcacc aaaatattgt tttcttcacc
     1441 aaccatcagt tcataggtcc attctcttag cgcaactaca gagaacaggg gcacaaacag
     1501 gcaaaaaacg ggcacaacct caatggagtg atgcaacctg cctggagtaa atgatgacac
     1561 aaggcaattg acccacgcat gtatctatct cattttctta caccttctat taccttctgc
     1621 tctctctgat ttggaaaaag ctgaaaaaaa aggttgaaac cagttccctg aaattattcc
     1681 cctacttgac taataagtat ataaagacgg taggtattga ttgtaattct gtaaatctat
     1741 ttcttaaact tcttaaattc tacttttata gttagtcttt tttttagttt taaaacacca
     1801 gaacttagtt tcgacggatt ctagaactag tggatccccc gggacaaaat gtcttctctt
     1861 agtagagagc tcgttttctt gatcttacag tttctcgatg aagagaagtt taaagagact
     1921 gttcacaagc ttgaacaaga atctgggttt ttcttcaata tgaagtattt tgaggatgag
     1981 gttcacaatg gaaattggga tgaggttgag aagtatctct ctggtttcac taaagtagat
     2041 gataatagat actctatgaa gatattcttc gagataagaa agcagaagta tctcgaggct
     2101 ttggataagc atgatcgtcc caaggctgtg gatatactag tgaaggatgg tggaccaggt
     2161 ggtggacatc aaacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat
     2221 atcaatatat taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa
     2281 catatccagt cactatggcg gccgcattag gcaccccagg ctttacactt tatgcttccg
     2341 gctcgtataa tgtgtggatt ttgagttagg atccgtcgag attttcagga gctaaggaag
     2401 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     2461 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     2521 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     2581 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag
     2641 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     2701 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     2761 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     2821 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     2881 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     2941 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtc tgtgatggct
     3001 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     3061 cgtaaacgcc gcgtggatcc ggcttactaa aagccagata acagtatgcg tatttgcgcg
     3121 ctgatttttg cggtataaga atatatactg atatgtatac ccgaagtatg tcaaaaagag
     3181 gtatgctatg aagcagcgta ttacagtgac agttgacagc gacagctatc agttgctcaa
     3241 ggcatatatg atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa tgaagcccgt
     3301 cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag ggatggctga ggtcgcccgg
     3361 tttattgaaa tgaacggctc ttttgctgac gagaacaggg gctggtgaaa tgcagtttaa
     3421 ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat
     3481 tattgacacg cccgggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga
     3541 taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat
     3601 gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg ctgatctcag
     3661 ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc
     3721 aggctccctt atacacagcc agtctgcagg tcgaccatag tgactggata tgttgtgttt
     3781 tacagtatta tgtagtctgt tttttatgca aaatctaatt taatatattg atatttatat
     3841 cattttacgt ttctcgttca gctttcttgt acaaagtggt tcgatgtcat gtaattagtt
     3901 atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt
     3961 agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg
     4021 ttatttatat ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt
     4081 atactgaaaa ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgtgcaggc
     4141 cttttgaaaa gcaagcataa aagatctaaa cataaaatct gtaaaataac aagatgtaaa
     4201 gataatgcta aatcatttgg ctttttgatt gattgtacag gaaaatatac atcgcagggg
     4261 gttgactttt accatttcac cgcaatggaa tcaaacttgt tgaagagaat gttcacaggc
     4321 gcatacgcta caatgacccg attcttgcta gccttttctc ggtcttgcaa acaaccgccg
     4381 gcagcttagt atataaatac acatgtacat acctctctcc gtatcctcgt aatcattttc
     4441 ttgtatttat cgtcttttcg ctgtaaaaac tttatcacac ttatctcaaa tacacttatt
     4501 aaccgctttt actattatct tctacgctga cagtaatatc aaacagtgac acatattaaa
     4561 cacagtggtt tctttgcata aacaccatgt ttaaaccatg gtcatagctg tttcctgtgt
     4621 gaaattgtta tccgctcaca attccacaca acataggagc cggaagcata aagtgtaaag
     4681 cctggggtgc ctaatgagtg aggtaactca cattaattgc gttgcgctca ctgcccgctt
     4741 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4801 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4861 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4921 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4981 aaaaggccgc gttgctggcg tttttccata ggctcggccc ccctgacgag catcacaaaa
     5041 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgttcc
     5101 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5161 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     5221 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5281 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5341 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5401 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     5461 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5521 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5581 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5641 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5701 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5761 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5821 tagttgcctg actgcccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5881 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5941 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6001 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6061 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6121 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgaaaaaaag
     6181 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6241 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6301 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6361 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6421 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6481 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6541 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6601 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6661 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6721 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     6781 cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
