LOCUS       sequence-160274-        7239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector for EYFP fusions integrated at yeast Trp locus, 
            expression from the GPD promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7239)
  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 7239)
  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: 
            "10.1073/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: pGP4GY-cccdB
FEATURES             Location/Qualifiers
     source          1..7239
                     /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"
     CDS             join(1849..1851,1852..1854,1855..2565)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1894..1915)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2155..2174)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2502..2523
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    2590..2714
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2739..2744,2745..2762,2763..2769)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2823..3482
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2889..2908)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3827..4132
                     /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     4011..4030
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4173..4297)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      4304..4550
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4320..4338)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5018..5034)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5018..5034)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5031..5053)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5042..5058
                     /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(5066..5072,5073..5090,5091..5096))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5249..5266)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5420..6008)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6179..6970,6971..7039))
                     /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     6802..6821
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7040..7144)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7212..7230
                     /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 ggtgagcaag
     1861 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1921 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     1981 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2041 ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     2101 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     2161 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2221 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2281 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     2341 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     2401 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
     2461 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2521 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggtgg accaggtggt
     2581 ggacatcaaa caagtttgta caaaaaagct gaacgagaaa cgtaaaatga tataaatatc
     2641 aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat
     2701 atccagtcac tatggcggcc gcattaggca ccccaggctt tacactttat gcttccggct
     2761 cgtataatgt gtggattttg agttaggatc cgtcgagatt ttcaggagct aaggaagcta
     2821 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     2881 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     2941 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     3001 ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg
     3061 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     3121 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     3181 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     3241 agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg
     3301 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     3361 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtctgt gatggcttcc
     3421 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     3481 aaacgccgcg tggatccggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg
     3541 atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca aaaagaggta
     3601 tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt tgctcaaggc
     3661 atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga agcccgtcgt
     3721 ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt cgcccggttt
     3781 attgaaatga acggctcttt tgctgacgag aacaggggct ggtgaaatgc agtttaaggt
     3841 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
     3901 tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa
     3961 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
     4021 caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca
     4081 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg
     4141 ctcccttata cacagccagt ctgcaggtcg accatagtga ctggatatgt tgtgttttac
     4201 agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat
     4261 tttacgtttc tcgttcagct ttcttgtaca aagtggttcg atgtcatgta attagttatg
     4321 tcacgcttac attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga
     4381 caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta
     4441 tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata
     4501 ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg tgcaggcctt
     4561 ttgaaaagca agcataaaag atctaaacat aaaatctgta aaataacaag atgtaaagat
     4621 aatgctaaat catttggctt tttgattgat tgtacaggaa aatatacatc gcagggggtt
     4681 gacttttacc atttcaccgc aatggaatca aacttgttga agagaatgtt cacaggcgca
     4741 tacgctacaa tgacccgatt cttgctagcc ttttctcggt cttgcaaaca accgccggca
     4801 gcttagtata taaatacaca tgtacatacc tctctccgta tcctcgtaat cattttcttg
     4861 tatttatcgt cttttcgctg taaaaacttt atcacactta tctcaaatac acttattaac
     4921 cgcttttact attatcttct acgctgacag taatatcaaa cagtgacaca tattaaacac
     4981 agtggtttct ttgcataaac accatgttta aaccatggtc atagctgttt cctgtgtgaa
     5041 attgttatcc gctcacaatt ccacacaaca taggagccgg aagcataaag tgtaaagcct
     5101 ggggtgccta atgagtgagg taactcacat taattgcgtt gcgctcactg cccgctttcc
     5161 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5221 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5281 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5341 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5401 aggccgcgtt gctggcgttt ttccataggc tcggcccccc tgacgagcat cacaaaaatc
     5461 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgttccccc
     5521 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5581 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     5641 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5701 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5761 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5821 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     5881 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5941 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6001 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6061 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6121 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6181 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6241 ttgcctgact gcccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6301 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6361 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6421 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6481 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6541 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtga aaaaaagcgg
     6601 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6661 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6721 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6781 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6841 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6901 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6961 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7021 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7081 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7141 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     7201 taacctataa aaataggcgt atcacgaggc cctttcgtc
//
