LOCUS       sequence-388940-        5750 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GAL4 DNA-binding domain (BD)-ubiquitin fusion protein 
            in yeast (yeast two-hybrid assays).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5750)
  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 5750)
  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: pGBT9-ubiquitin
FEATURES             Location/Qualifiers
     source          1..5750
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(301..1092,1093..1161))
                     /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     924..943
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1162..1266)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1334..1352
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1390..1412)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1512..1531
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      1548..2712
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        2717..3118
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             3146..3586
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             3596..3823
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             3596..3823
                     /codon_start=1
                     /gene="Homo sapiens UBB"
                     /product="human ubiquitin B"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     terminator      3860..4047
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3915..3939)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(4100..4774)
                     /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(4775..4876)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(4890..4906)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4890..4906)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4903..4925)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4914..4930
                     /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(4938..4944,4945..4962,4963..4968))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4983..5004
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5121..5138)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5292..5750,1..130))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5372..5391)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
       61 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
      121 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
      181 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
      241 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
      301 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
      361 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      421 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
      481 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      541 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      601 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      661 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      721 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      781 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      841 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      901 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
      961 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     1021 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     1081 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     1141 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     1201 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     1261 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     1321 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     1381 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     1441 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     1501 gcttaactat gcggcatcag agcagattgt actgagagtg caccataacg catttaagca
     1561 taaacacgca ctatgccgtt cttctcatgt atatatatat acaggcaaca cgcagatata
     1621 ggtgcgacgt gaacagtgag ctgtatgtgc gcagctcgcg ttgcattttc ggaagcgctc
     1681 gttttcggaa acgctttgaa gttcctattc cgaagttcct attctctagc tagaaagtat
     1741 aggaacttca gagcgctttt gaaaaccaaa agcgctctga agacgcactt tcaaaaaacc
     1801 aaaaacgcac cggactgtaa cgagctacta aaatattgcg aataccgctt ccacaaacat
     1861 tgctcaaaag tatctctttg ctatatatct ctgtgctata tccctatata acctacccat
     1921 ccacctttcg ctccttgaac ttgcatctaa actcgacctc tacatttttt atgtttatct
     1981 ctagtattac tctttagaca aaaaaattgt agtaagaact attcatagag tgaatcgaaa
     2041 acaatacgaa aatgtaaaca tttcctatac gtagtatata gagacaaaat agaagaaacc
     2101 gttcataatt ttctgaccaa tgaagaatca tcaacgctat cactttctgt tcacaaagta
     2161 tgcgcaatcc acatcggtat agaatataat cggggatgcc tttatcttga aaaaatgcac
     2221 ccgcagcttc gctagtaatc agtaaacgcg ggaagtggag tcaggctttt tttatggaag
     2281 agaaaataga caccaaagta gccttcttct aaccttaacg gacctacagt gcaaaaagtt
     2341 atcaagagac tgcattatag agcgcacaaa ggagaaaaaa agtaatctaa gatgctttgt
     2401 tagaaaaata gcgctctcgg gatgcatttt tgtagaacaa aaaagaagta tagattcttt
     2461 gttggtaaaa tagcgctctc gcgttgcatt tctgttctgt aaaaatgcag ctcagattct
     2521 ttgtttgaaa aattagcgct ctcgcgttgc atttttgttt tacaaaaatg aagcacagat
     2581 tcttcgttgg taaaatagcg ctttcgcgtt gcatttctgt tctgtaaaaa tgcagctcag
     2641 attctttgtt tgaaaaatta gcgctctcgc gttgcatttt tgttctacaa aatgaagcac
     2701 agatgcttcg ttgcttgcat gcaacttctt ttcttttttt ttcttttctc tctcccccgt
     2761 tgttgtctca ccatatccgc aatgacaaaa aaatgatgga agacactaaa ggaaaaaatt
     2821 aacgacaaag acagcaccaa cagatgtcgt tgttccagag ctgatgaggg gtatctcgaa
     2881 gcacacgaaa ctttttcctt ccttcattca cgcacactac tctctaatga gcaacggtat
     2941 acggccttcc ttccagttac ttgaatttga aataaaaaaa agtttgctgt cttgctatca
     3001 agtataaata gacctgcaat tattaatctt ttgtttcctc gtcattgttc tcgttccctt
     3061 tcttccttgt ttctttttct gcacaatatt tcaagctata ccaagcatac aatcaactcc
     3121 aagcttgaag caagcctcct gaaagatgaa gctactgtct tctatcgaac aagcatgcga
     3181 tatttgccga cttaaaaagc tcaagtgctc caaagaaaaa ccgaagtgcg ccaagtgtct
     3241 gaagaacaac tgggagtgtc gctactctcc caaaaccaaa aggtctccgc tgactagggc
     3301 acatctgaca gaagtggaat caaggctaga aagactggaa cagctatttc tactgatttt
     3361 tcctcgagaa gaccttgaca tgattttgaa aatggattct ttacaggata taaaagcatt
     3421 gttaacagga ttatttgtac aagataatgt gaataaagat gccgtcacag atagattggc
     3481 ttcagtggag actgatatgc ctctaacatt gagacagcat agaataagtg cgacatcatc
     3541 atcggaagag agtagtaaca aaggtcaaag acagttgact gtatcgccgg aattcatgca
     3601 gatcttcgtg aagaccctga ccggcaagac catcaccctg gaagtggagc ccagtgacac
     3661 catcgaaaat gtgaaggcca agatccagga taaagaaggc atccctcccg accagcagag
     3721 gctcatcttt gcaggcaagc agctggaaga tggccgcact ctttctgact acaacatcca
     3781 gaaggagtcg accctgcacc tggtcctgcg tctgagaggt ggttgaggat ccgtcgacct
     3841 gcagccaagc taattccggg cgaatttctt atgatttatg atttttatta ttaaataagt
     3901 tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt
     3961 cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat agcatgaggt
     4021 cgctcttatt gaccacacct ctaccggcat gccggcaagt gcacaaacaa tacttaaata
     4081 aatactactc agtaataacc tatttcttag catttttgac gaaatttgct attttgttag
     4141 agtcttttac accatttgtc tccacacctc cgcttacatc aacaccaata acgccattta
     4201 atctaagcgc atcaccaaca ttttctggcg tcagtccacc agctaacata aaatgtaagc
     4261 tttcggggct ctcttgcctt ccaacccagt cagaaatcga gttccaatcc aaaagttcac
     4321 ctgtcccacc tgcttctgaa tcaaacaagg gaataaacga atgaggtttc tgtgaagctg
     4381 cactgagtag tatgttgcag tcttttggaa atacgagtct tttaataact ggcaaaccga
     4441 ggaactcttg gtattcttgc cacgactcat ctccatgcag ttggacgata tcaatgccgt
     4501 aatcattgac cagagccaaa acatcctcct taggttgatt acgaaacacg ccaaccaagt
     4561 atttcggagt gcctgaacta tttttatatg cttttacaag acttgaaatt ttccttgcaa
     4621 taaccgggtc aattgttctc tttctattgg gcacacatat aatacccagc aagtcagcat
     4681 cggaatctag agcacattct gcggcctctg tgctctgcaa gccgcaaact ttcaccaatg
     4741 gaccagaact acctgtgaaa ttaataacag acatactcca agctgccttt gtgtgcttaa
     4801 tcacgtatac tcacgtgctc aatagtcacc aatgccctcc ctcttggccc tctccttttc
     4861 ttttttcgac cgaattaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat
     4921 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4981 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5041 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5101 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5161 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5221 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5281 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5341 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5401 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5461 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5521 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5581 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5641 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5701 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
//
