LOCUS       sequence-364071-        7259 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GAL4 fusion of Capsaspora NF-kB Rel Homology Domain 
            sequences in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7259)
  AUTHORS   Williams LM, Sridhar S, Samaroo J, Peart J, Adindu EK, Addanki A, 
            DiRusso CJ, Aguirre Carrion PJ, Rodriguez-Sastre N, Siggers T, 
            Gilmore TD
  TITLE     Comparison of NF-kappaB from the protists Capsaspora owczarzaki and 
            Acanthoeca spectabilis reveals extensive evolutionary 
            diversification of this transcription factor.
  JOURNAL   Commun Biol. 2021 Dec 16;4(1):1404. doi: 10.1038/s42003-021-02924-2.
  PUBMED    34916615
REFERENCE   2  (bases 1 to 7259)
  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"; journalName: "Commun 
            Biol."; date: "2021-12-16"; volume: "4(1):1404. doi"; pages: " 
            10.1038/s42003-021-02924-2"
COMMENT     Sequence Label: pGB-Co-RHD
FEATURES             Location/Qualifiers
     source          1..7259
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1906..2093
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1961..1985)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2146..2820)
                     /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(2821..2922)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(2936..2952)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2936..2952)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2949..2971)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2960..2976
                     /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(2984..2990,2991..3008,3009..3014))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3029..3050
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3167..3184)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3338..3926)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3418..3437)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4097..4888,4889..4957))
                     /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     4720..4739
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4958..5062)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5130..5148
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5186..5208)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5308..5327
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      5344..6508
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        6513..6914
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             join(6942..7259,1..123)
                     /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"
ORIGIN
        1 gtcacagata gattggcttc agtggagact gatatgcctc taacattgag acagcataga
       61 ataagtgcga catcatcatc ggaagagagt agtaacaaag gtcaaagaca gttgactgta
      121 tcgccggaat tcgacctttc tgaactttcc ggatgggacc cgaatttgtc cttgcaggag
      181 cacactgcaa atctgctcgc tatggatgat tcaacaatgg cagctatgat actgcattct
      241 gatggactgt cattgttcag agatatgggt ctgtacaatt ccgtctctac gagcctggat
      301 attagcggta ttccaaaatt tccgccaccg ccacagcagc cgcaacttgc acaagcgccg
      361 cctcggagag gtcacaacca atccagcagt agcgatagcc atagcacacc atcacccggt
      421 tccgtgctct tctccccgag cccggcttct caagatatga gcctccagag tccggagctg
      481 ggcctcgttg ggcttactgg caacagtagg ctggcgtcaa cgtcagaggc cgatgatatt
      541 ctgcttgcga atatactggg gcacccagta cggtcagtta gtgcaaatac atcaatggtt
      601 ggtcttccgg atgacctggg attctccccc acgaccggaa tggatctcac aatcacggcg
      661 acgagtccag cgacggcaga ctcatccgcg tcagctacag ccttccctgc ggcgtccccg
      721 atagcttcac ctagcgtctc caccagctcc gggcctgtaa ctgttggcgg aaccagcgcg
      781 gccttggccg cattgactcc ggacagaatt aataggcttc ttagcgcagc cgaggcggca
      841 gccgaagggg cggccaccct cgttgaggac cttcttatgg tcacggagga gcccgcgcag
      901 ttcgcccgat tccgctacat gagcgagcag cgagagagat ctttggccgg cgagaactcc
      961 ttccccacgc ttatggtaaa ccctaagtat gccagagtgg tccccgagat ggccctcgta
     1021 actgctgtcc tggtcacaaa aatgccggat ccccacactg ggaggcagca gaagcattgg
     1081 catcaccttg ggggcatacc tgcggctcct ctggaagggc ctcaacgcat agcaaggttc
     1141 gacaacattg ctgtaataat ggacaaggcg aacaataagg ataaggacaa atcaaaggca
     1201 cccgtaagat ctaaagacga tcagcgatgt gtcaggataa tgtttgaact tgtctttgtg
     1261 tcaggtaata cacagttcta tggcagggca atcagccaac caatctacaa cgccaaattg
     1321 gccatcacaa agattagtca tagcagcggg cccgtgaccg gtggtaacga ggttatcatg
     1381 ttgtgcagta agatcagaaa aggcgtgacc ggggtacgga tgaccgatcc aacccaatgg
     1441 tcagtccagg cacctagcgg ctctgcgtgg gagttgaatc cacaaacgct gaaggccgac
     1501 tgcaatgtac cgggcgctaa tctttttttc catcatcaat atgcagtcgt cctgacgctg
     1561 ccgccctacc acacgcagac cataacagcc cctgtcaccg tcaggatctc aatcttggac
     1621 actgacgatg agacagagag tcaatacgtt gagtatacct atttgccggc cgaagcggcg
     1681 gtgcgcaacg cagagctggc ggctcggaaa cggagacggg atgattcaat gagagatttt
     1741 atggaccgct ttgatgggtc cgacggtgga aatggctctg ggagtggccg aggtaacaac
     1801 ggaggacacg atggctctga cgctaacaat aacggccgag gcggtggcgg tgggagttct
     1861 tcaagtaaag gcgggtaact cgacctgcag ccaagctaat tccgggcgaa tttcttatga
     1921 tttatgattt ttattattaa ataagttata aaaaaaataa gtgtatacaa attttaaagt
     1981 gactcttagg ttttaaaacg aaaattctta ttcttgagta actctttcct gtaggtcagg
     2041 ttgctttctc aggtatagca tgaggtcgct cttattgacc acacctctac cggcatgccg
     2101 gcaagtgcac aaacaatact taaataaata ctactcagta ataacctatt tcttagcatt
     2161 tttgacgaaa tttgctattt tgttagagtc ttttacacca tttgtctcca cacctccgct
     2221 tacatcaaca ccaataacgc catttaatct aagcgcatca ccaacatttt ctggcgtcag
     2281 tccaccagct aacataaaat gtaagctttc ggggctctct tgccttccaa cccagtcaga
     2341 aatcgagttc caatccaaaa gttcacctgt cccacctgct tctgaatcaa acaagggaat
     2401 aaacgaatga ggtttctgtg aagctgcact gagtagtatg ttgcagtctt ttggaaatac
     2461 gagtctttta ataactggca aaccgaggaa ctcttggtat tcttgccacg actcatctcc
     2521 atgcagttgg acgatatcaa tgccgtaatc attgaccaga gccaaaacat cctccttagg
     2581 ttgattacga aacacgccaa ccaagtattt cggagtgcct gaactatttt tatatgcttt
     2641 tacaagactt gaaattttcc ttgcaataac cgggtcaatt gttctctttc tattgggcac
     2701 acatataata cccagcaagt cagcatcgga atctagagca cattctgcgg cctctgtgct
     2761 ctgcaagccg caaactttca ccaatggacc agaactacct gtgaaattaa taacagacat
     2821 actccaagct gcctttgtgt gcttaatcac gtatactcac gtgctcaata gtcaccaatg
     2881 ccctccctct tggccctctc cttttctttt ttcgaccgaa ttaattcgta atcatgtcat
     2941 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3001 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3061 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3121 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3181 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3241 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3301 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3361 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3421 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3481 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3541 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3601 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3661 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3721 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3781 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3841 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3901 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3961 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4021 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4081 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4141 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4201 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4261 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4321 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4381 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4441 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4501 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4561 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4621 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4681 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4741 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4801 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4861 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4921 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4981 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5041 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     5101 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     5161 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     5221 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     5281 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     5341 ataacgcatt taagcataaa cacgcactat gccgttcttc tcatgtatat atatatacag
     5401 gcaacacgca gatataggtg cgacgtgaac agtgagctgt atgtgcgcag ctcgcgttgc
     5461 attttcggaa gcgctcgttt tcggaaacgc tttgaagttc ctattccgaa gttcctattc
     5521 tctagctaga aagtatagga acttcagagc gcttttgaaa accaaaagcg ctctgaagac
     5581 gcactttcaa aaaaccaaaa acgcaccgga ctgtaacgag ctactaaaat attgcgaata
     5641 ccgcttccac aaacattgct caaaagtatc tctttgctat atatctctgt gctatatccc
     5701 tatataacct acccatccac ctttcgctcc ttgaacttgc atctaaactc gacctctaca
     5761 ttttttatgt ttatctctag tattactctt tagacaaaaa aattgtagta agaactattc
     5821 atagagtgaa tcgaaaacaa tacgaaaatg taaacatttc ctatacgtag tatatagaga
     5881 caaaatagaa gaaaccgttc ataattttct gaccaatgaa gaatcatcaa cgctatcact
     5941 ttctgttcac aaagtatgcg caatccacat cggtatagaa tataatcggg gatgccttta
     6001 tcttgaaaaa atgcacccgc agcttcgcta gtaatcagta aacgcgggaa gtggagtcag
     6061 gcttttttta tggaagagaa aatagacacc aaagtagcct tcttctaacc ttaacggacc
     6121 tacagtgcaa aaagttatca agagactgca ttatagagcg cacaaaggag aaaaaaagta
     6181 atctaagatg ctttgttaga aaaatagcgc tctcgggatg catttttgta gaacaaaaaa
     6241 gaagtataga ttctttgttg gtaaaatagc gctctcgcgt tgcatttctg ttctgtaaaa
     6301 atgcagctca gattctttgt ttgaaaaatt agcgctctcg cgttgcattt ttgttttaca
     6361 aaaatgaagc acagattctt cgttggtaaa atagcgcttt cgcgttgcat ttctgttctg
     6421 taaaaatgca gctcagattc tttgtttgaa aaattagcgc tctcgcgttg catttttgtt
     6481 ctacaaaatg aagcacagat gcttcgttgc ttgcatgcaa cttcttttct ttttttttct
     6541 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat gatggaagac
     6601 actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt ccagagctga
     6661 tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca cactactctc
     6721 taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata aaaaaaagtt
     6781 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca
     6841 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa
     6901 gcatacaatc aactccaagc ttgaagcaag cctcctgaaa gatgaagcta ctgtcttcta
     6961 tcgaacaagc atgcgatatt tgccgactta aaaagctcaa gtgctccaaa gaaaaaccga
     7021 agtgcgccaa gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa accaaaaggt
     7081 ctccgctgac tagggcacat ctgacagaag tggaatcaag gctagaaaga ctggaacagc
     7141 tatttctact gatttttcct cgagaagacc ttgacatgat tttgaaaatg gattctttac
     7201 aggatataaa agcattgtta acaggattat ttgtacaaga taatgtgaat aaagatgcc
//
