LOCUS       sequence-388865-        8187 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GAL4 transcriptional activation domain (AD)-Rabex-5 
            fusion protein in yeast (yeast two-hybrid assays).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8187)
  AUTHORS   Mattera R, Tsai YC, Weissman AM, Bonifacino JS
  TITLE     The Rab5 guanine nucleotide exchange factor Rabex-5 binds ubiquitin 
            (Ub) and functions as a Ub ligase through an atypical Ub-interacting
            motif and a zinc finger domain.
  JOURNAL   J Biol Chem. 2006 Mar 10;281(10):6874-83. doi: 
            10.1074/jbc.M509939200. Epub 2006 Jan 5.
  PUBMED    16407276
REFERENCE   2  (bases 1 to 8187)
  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.M509939200"; journalName: "J Biol Chem"; date: 
            "2006-03-10- 10"; volume: "281"; issue: "10"; pages: "6874-83"
COMMENT     Sequence Label: pGAD424-Rabex-5
FEATURES             Location/Qualifiers
     source          1..8187
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      370..557
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(425..449)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(678..1772)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1773..2178)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(2220..2236)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2220..2236)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2233..2255)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2244..2260
                     /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(2268..2274,2275..2292,2293..2298))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2313..2334
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2451..2468)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2622..3210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2702..2721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3381..4172,4173..4241))
                     /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     4004..4023
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4242..4346)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4414..4432
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4470..4492)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4592..4611
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      4628..5792
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        5797..6198
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             6244..6264
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6280..6621
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
ORIGIN
        1 tcgtagatac tgaaaaaccc cgcaagttca cttcaactgt gcatcgtgca ccatctcaat
       61 ttctttcatt tatacatcgt tttgccttct tttatgtaac tatactcctc taagtttcaa
      121 tcttggccat gtaacctctg atctatagaa ttttttaaat gactagaatt aatgcccatc
      181 ttttttttgg acctaaattc ttcatgaaaa tatattacga gggcttattc agaagctttg
      241 gacttcttcg ccagaggttt ggtcaagtct ccaatcaagg ttgtcggctt gtctaccttg
      301 ccagaaattt acgaaaagat ggaaaagggt caaatcgttg gtagatacgt tgttgacact
      361 tctaaataag cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa
      421 ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg
      481 agtaactctt tcctgtaggt caggttgctt tctcaggtat agcatgaggt cgctcttatt
      541 gaccacacct ctaccggcat gccggtcgaa attcccctac cctatgaaca tattccattt
      601 tgtaatttcg tgtcgtttct attatgaatt tcatttataa agtttatgta caaatatcat
      661 aaaaaaagag aatcttttta agcaaggatt ttcttaactt cttcggcgac agcatcaccg
      721 acttcggtgg tactgttgga accacctaaa tcaccagttc tgatacctgc atccaaaacc
      781 tttttaactg catcttcaat ggccttacct tcttcaggca agttcaatga caatttcaac
      841 atcattgcag cagacaagat agtggcgata gggttgacct tattctttgg caaatctgga
      901 gcagaaccgt ggcatggttc gtacaaacca aatgcggtgt tcttgtctgg caaagaggcc
      961 aaggacgcag atggcaacaa acccaaggaa cctgggataa cggaggcttc atcggagatg
     1021 atatcaccaa acatgttgct ggtgattata ataccattta ggtgggttgg gttcttaact
     1081 aggatcatgg cggcagaatc aatcaattga tgttgaacct tcaatgtagg aaattcgttc
     1141 ttgatggttt cctccacagt ttttctccat aatcttgaag aggccaaaac attagcttta
     1201 tccaaggacc aaataggcaa tggtggctca tgttgtaggg ccatgaaagc ggccattctt
     1261 gtgattcttt gcacttctgg aacggtgtat tgttcactat cccaagcgac accatcacca
     1321 tcgtcttcct ttctcttacc aaagtaaata cctcccacta attctctgac aacaacgaag
     1381 tcagtacctt tagcaaattg tggcttgatt ggagataagt ctaaaagaga gtcggatgca
     1441 aagttacatg gtcttaagtt ggcgtacaat tgaagttctt tacggatttt tagtaaacct
     1501 tgttcaggtc taacactacc tgtaccccat ttaggaccac ccacagcacc taacaaaacg
     1561 gcatcagcct tcttggaggc ttccagcgcc tcatctggaa gtggaacacc tgtagcatcg
     1621 atagcagcac caccaattaa atgattttcg aaatcgaact tgacattgga acgaacatca
     1681 gaaatagctt taagaacctt aatggcttcg gctgtgattt cttgaccaac gtggtcacct
     1741 ggcaaaacga cgatcttctt aggggcagac attagaatgg tatatccttg aaatatatat
     1801 atatattgct gaaatgtaaa aggtaagaaa agttagaaag taagacgatt gctaaccacc
     1861 tattggaaaa aacaataggt ccttaaataa tattgtcaac ttcaagtatt gtgatgcaag
     1921 catttagtca tgaacgcttc tctattctat atgaaaagcc ggttccggcg ctctcacctt
     1981 tcctttttct cccaattttt cagttgaaaa aggtatatgc gtcaggcgac ctctgaaatt
     2041 aacaaaaaat ttccagtcat cgaatttgat tctgtgcgat agcgcccctg tgtgttctcg
     2101 ttatgttgag gaaaaaaata atggttgcta agagattcga actcttgcat cttacgatac
     2161 ctgagtattc ccacagttgg gggatctcga ctctagctag aggatcaatt cgtaatcatg
     2221 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     2281 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2341 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2401 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2461 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2521 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2581 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2641 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2701 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2761 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2821 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2881 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2941 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3001 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3061 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3121 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3181 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3241 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3301 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3361 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3421 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3481 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3541 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3601 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3661 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3721 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3781 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3841 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3901 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3961 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4021 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4081 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4141 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4201 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4261 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4321 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     4381 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     4441 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     4501 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     4561 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     4621 caccataacg catttaagca taaacacgca ctatgccgtt cttctcatgt atatatatat
     4681 acaggcaaca cgcagatata ggtgcgacgt gaacagtgag ctgtatgtgc gcagctcgcg
     4741 ttgcattttc ggaagcgctc gttttcggaa acgctttgaa gttcctattc cgaagttcct
     4801 attctctagc tagaaagtat aggaacttca gagcgctttt gaaaaccaaa agcgctctga
     4861 agacgcactt tcaaaaaacc aaaaacgcac cggactgtaa cgagctacta aaatattgcg
     4921 aataccgctt ccacaaacat tgctcaaaag tatctctttg ctatatatct ctgtgctata
     4981 tccctatata acctacccat ccacctttcg ctccttgaac ttgcatctaa actcgacctc
     5041 tacatttttt atgtttatct ctagtattac tctttagaca aaaaaattgt agtaagaact
     5101 attcatagag tgaatcgaaa acaatacgaa aatgtaaaca tttcctatac gtagtatata
     5161 gagacaaaat agaagaaacc gttcataatt ttctgaccaa tgaagaatca tcaacgctat
     5221 cactttctgt tcacaaagta tgcgcaatcc acatcggtat agaatataat cggggatgcc
     5281 tttatcttga aaaaatgcac ccgcagcttc gctagtaatc agtaaacgcg ggaagtggag
     5341 tcaggctttt tttatggaag agaaaataga caccaaagta gccttcttct aaccttaacg
     5401 gacctacagt gcaaaaagtt atcaagagac tgcattatag agcgcacaaa ggagaaaaaa
     5461 agtaatctaa gatgctttgt tagaaaaata gcgctctcgg gatgcatttt tgtagaacaa
     5521 aaaagaagta tagattcttt gttggtaaaa tagcgctctc gcgttgcatt tctgttctgt
     5581 aaaaatgcag ctcagattct ttgtttgaaa aattagcgct ctcgcgttgc atttttgttt
     5641 tacaaaaatg aagcacagat tcttcgttgg taaaatagcg ctttcgcgtt gcatttctgt
     5701 tctgtaaaaa tgcagctcag attctttgtt tgaaaaatta gcgctctcgc gttgcatttt
     5761 tgttctacaa aatgaagcac agatgcttcg ttgcttgcat gcaacttctt ttcttttttt
     5821 ttcttttctc tctcccccgt tgttgtctca ccatatccgc aatgacaaaa aaatgatgga
     5881 agacactaaa ggaaaaaatt aacgacaaag acagcaccaa cagatgtcgt tgttccagag
     5941 ctgatgaggg gtatctcgaa gcacacgaaa ctttttcctt ccttcattca cgcacactac
     6001 tctctaatga gcaacggtat acggccttcc ttccagttac ttgaatttga aataaaaaaa
     6061 agtttgctgt cttgctatca agtataaata gacctgcaat tattaatctt ttgtttcctc
     6121 gtcattgttc tcgttccctt tcttccttgt ttctttttct gcacaatatt tcaagctata
     6181 ccaagcatac aatcaactcc aagctttgca aagatggata aagcggaatt aattcccgag
     6241 cctccaaaaa agaagagaaa ggtcgaattg ggtaccgccg ccaattttaa tcaaagtggg
     6301 aatattgctg atagctcatt gtccttcact ttcactaaca gtagcaacgg tccgaacctc
     6361 ataacaactc aaacaaattc tcaagcgctt tcacaaccaa ttgcctcctc taacgttcat
     6421 gataacttca tgaataatga aatcacggct agtaaaattg atgatggtaa taattcaaaa
     6481 ccactgtcac ctggttggac ggaccaaact gcgtataacg cgtttggaat cactacaggg
     6541 atgtttaata ccactacaat ggatgatgta tataactatc tattcgatga tgaagatacc
     6601 ccaccaaacc caaaaaaaga gatcgaattc ccggggatcg gccgctctag ccctatgagc
     6661 cttaagtctg aacgccgagg aattcatgtg gatcagtcgg agctcctatg caagaaagga
     6721 tgtggttact acggcaaccc tgcttggcag ggtttctgct ccaagtgctg gagagaggag
     6781 taccacaaag ccaggcagaa gcagattcag gaggactggg aactggcaga gcgactgcag
     6841 cgggaggaag aagaggcctt tgccagcagt cagagcagcc aaggtgccca atccctcaca
     6901 ttttccaagt ttgaagagaa gaaaaccaat gagaagaccc gcaaggttac cacagtgaag
     6961 aaattcttca gtgcttcatc cagggttgga gcaaagaagg cagaaattca ggaagcaaaa
     7021 gctcccagtc cttccataaa ccggcaaacc agcattgaaa cggatagagt gtctaaagag
     7081 ttcatagaat ttctcaagac cttccacaag acaggccaag aaatctataa gcagaccaag
     7141 ctgtttttgg aagcaatgca ttacaaaagg gacttaagta ttgaggagca gtcagagtgt
     7201 actcaggatt tttaccagaa tgtggctgaa agaatgcaga ctcgtggaaa agtgcctcca
     7261 gaaagagttg agaagataat ggatcagatt gaaaagtaca tcatgactcg tctctataaa
     7321 tatgtgttct gtccagaaac tactgatgat gagaagaaag atcttgctat tcagaagagg
     7381 atcagagccc tgcactgggt tacacctcag atgctttgtg tccctgttaa tgaagaaatt
     7441 ccagaagtgt ccgatatggt ggtgaaagca attacagata tcattgagat ggattcgaaa
     7501 cgtgtgcccc gggataagtt ggcctgcatc accaagtgca gcaagcacat cttcaatgcc
     7561 atcaagatca ccaagaatga gccggcttca gccgacgact tcttacccac actcatctac
     7621 attgtcttga agggcaaccc cccacgcctt cagtccaaca tccagtacat cacccgcttc
     7681 tgcaacccaa gccgactgat gacgggtgag gatggctact atttcaccaa tctgtgctgt
     7741 gccgtggctt tcattgagaa attagatgct cagtctttga atttaagtca ggaagatttt
     7801 gatcgataca tgtctggcca gacttctccc aggaagcagg aatctgagaa ttggtctcct
     7861 gatgcgtgct taggtgtgaa gcagatgtat aagaatttgg acctcctgtc tcagttgaat
     7921 gaacgacagg aaaggattat gaatgaagcc aagaaacttg agaaagacct catagattgg
     7981 acggatggaa tcgcaaaaga ggttcaagac attgttgaga aatacccact tgaaattaga
     8041 cccccaaatc agtctgtagc agctattgac tctgaaaatg ttgaaaatga taaacttcct
     8101 ccaccgttgc aacctcaagt ttatgcagga taggctcaga ctacccgggg gaatcactag
     8161 atccgtcgac ctgcagagat ctatgaa
//
