LOCUS       sequence-357180-        7562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GAL4 fusion of Capsaspora NF-kB C-terminal sequences in 
            yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7562)
  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 7562)
  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-Cterm
FEATURES             Location/Qualifiers
     source          1..7562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             336..776
                     /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"
     terminator      2862..3049
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2917..2941)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(3102..3776)
                     /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(3777..3878)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3892..3908)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3892..3908)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3905..3927)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3916..3932
                     /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(3940..3946,3947..3964,3965..3970))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3985..4006
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4123..4140)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4294..4882)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4374..4393)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5053..5844,5845..5913))
                     /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     5676..5695
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5914..6018)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6086..6104
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6142..6164)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6264..6283
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      6300..7464
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        join(7469..7562,1..308)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
ORIGIN
        1 ggaaaaaatt aacgacaaag acagcaccaa cagatgtcgt tgttccagag ctgatgaggg
       61 gtatctcgaa gcacacgaaa ctttttcctt ccttcattca cgcacactac tctctaatga
      121 gcaacggtat acggccttcc ttccagttac ttgaatttga aataaaaaaa agtttgctgt
      181 cttgctatca agtataaata gacctgcaat tattaatctt ttgtttcctc gtcattgttc
      241 tcgttccctt tcttccttgt ttctttttct gcacaatatt tcaagctata ccaagcatac
      301 aatcaactcc aagcttgaag caagcctcct gaaagatgaa gctactgtct tctatcgaac
      361 aagcatgcga tatttgccga cttaaaaagc tcaagtgctc caaagaaaaa ccgaagtgcg
      421 ccaagtgtct gaagaacaac tgggagtgtc gctactctcc caaaaccaaa aggtctccgc
      481 tgactagggc acatctgaca gaagtggaat caaggctaga aagactggaa cagctatttc
      541 tactgatttt tcctcgagaa gaccttgaca tgattttgaa aatggattct ttacaggata
      601 taaaagcatt gttaacagga ttatttgtac aagataatgt gaataaagat gccgtcacag
      661 atagattggc ttcagtggag actgatatgc ctctaacatt gagacagcat agaataagtg
      721 cgacatcatc atcggaagag agtagtaaca aaggtcaaag acagttgact gtatcgccgg
      781 aattcgatgg gtccgacggt ggaaatggct ctgggagtgg ccgaggtaac aacggaggac
      841 acgatggctc tgacgctaac aataacggcc gaggcggtgg cggtgggagt tcttcaagta
      901 aaggcgggga cgaaccgttt aattttaatt cactcattcc aatgcaccaa cacaagcttc
      961 atcaactggc gctctccacg gtaagggctg tgcaagggtt cgcggccagt ggggatgcac
     1021 ggtatctgtt ggccttgcat agacagttgc ttgcagctcc gaacgaaaac ggtgactccc
     1081 cgctgcacac agcagtagca cagggtaatc tcagatctac tatggcattg ctcccattgc
     1141 ttgcggcaga agatctccaa agtgtgaacg acatggggga aactgtcttg catagcgctg
     1201 taattgagaa aagggcagcc atagctcggc tgcttctggt cgctggcgca gatcttggcc
     1261 agagtaatgc acggaacttt aatcgaaaca gccttcacta ccttgcccgg catggagata
     1321 gggcgactgc aatggctgtg ttcggcgttt ttggcagtgc gcaagcccca ccagctaaca
     1381 cgaacacgcc cgcacaggca ccggcaggcg aaacgaagcc gaaaccagct gacttgagac
     1441 tgttggcaag gattcaggcg caggctataa aagcgctcct ggcctgtgaa ctcgaaactg
     1501 gggccactcc cgcccatttg gcgatacggg gtggccactg gcacgtcttt gaagcgtgtg
     1561 ctaagttggc ggcatcagca cccataccca aagccgctgg ttctcttctt tctatggtcg
     1621 ctgaaaagag tagtggacat agtctgttgc actcctgcgt cctggccaat aatgagcagg
     1681 ctgttcgact tcttataaat ctgggggcct ctggcaacgc tagggacttc gggaagaaca
     1741 cgccacttca cctggccgca cgacaaggac acatcggcat agccgcgctg ttggtagagg
     1801 caggggctac cctgagtctt aacgccgtta gccagactcc cctcgatgtt ttgacttcag
     1861 agggttccgg attgagcaga gatcagctcc gggcattggt agcagtcctc cgaggcgagc
     1921 tcaaatatgc cgacatgcgc ggccggccaa ccctccgaat gccgactcac gcggagctgc
     1981 atagcacggc ggccgcactc acttccgcct ccccaggggc agtctcactg gctgatttct
     2041 acgcgggcaa aaaggcctca aggagccccg cccctctcgg tgcttcctca agtcttctta
     2101 gtagtactgg agcttctgcg gctggcgcct ctgcgcccac gatagcggcg gtgcatgccg
     2161 cgagcgcaac accggtagag cgcaccagca tgaacaatga cgatgattac gttctccttg
     2221 agaaggacgc gccttaccct gttgagcagc agcctcacgg gaagcgcaac aagcattctc
     2281 accacagatt cacccggtcc agtcacggct cacaggataa agatgagctt aaaaaagata
     2341 aggatgaccc gaaaaaagaa aaagagccga aggaactgtc aaaattcaca ctgaaagagg
     2401 cgtttgtgga cggtacgaat ttttgggaac ttactcggaa attcgctggg aagaagaaaa
     2461 tggcttcagc aagtactggc gagatggagc cgttgagtcc ggaacgacca ctcagtccga
     2521 ccaatgcggg aagtggggct gcctcacctt ttaaccaagc taaggaacaa gttagccccg
     2581 gtgctgtacc tccaacaggt cttgagaagc ttgtcaacaa gctcatggac gcttcagagg
     2641 ctaccctctc aagtcaacca gctgaggccg tgacgcccga acagaaactg gctgaaaaac
     2701 ttgaaaaatt gggactggca cctgcgtcta ccactagtgc gccgccaccg cacccaaaag
     2761 ttgctgctct gaatgcgcag tctgtcgaag atgcgcggaa gacttctacc catgcccttt
     2821 actccgttga ttaactcgac ctgcagccaa gctaattccg ggcgaatttc ttatgattta
     2881 tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact
     2941 cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc
     3001 tttctcaggt atagcatgag gtcgctctta ttgaccacac ctctaccggc atgccggcaa
     3061 gtgcacaaac aatacttaaa taaatactac tcagtaataa cctatttctt agcatttttg
     3121 acgaaatttg ctattttgtt agagtctttt acaccatttg tctccacacc tccgcttaca
     3181 tcaacaccaa taacgccatt taatctaagc gcatcaccaa cattttctgg cgtcagtcca
     3241 ccagctaaca taaaatgtaa gctttcgggg ctctcttgcc ttccaaccca gtcagaaatc
     3301 gagttccaat ccaaaagttc acctgtccca cctgcttctg aatcaaacaa gggaataaac
     3361 gaatgaggtt tctgtgaagc tgcactgagt agtatgttgc agtcttttgg aaatacgagt
     3421 cttttaataa ctggcaaacc gaggaactct tggtattctt gccacgactc atctccatgc
     3481 agttggacga tatcaatgcc gtaatcattg accagagcca aaacatcctc cttaggttga
     3541 ttacgaaaca cgccaaccaa gtatttcgga gtgcctgaac tatttttata tgcttttaca
     3601 agacttgaaa ttttccttgc aataaccggg tcaattgttc tctttctatt gggcacacat
     3661 ataataccca gcaagtcagc atcggaatct agagcacatt ctgcggcctc tgtgctctgc
     3721 aagccgcaaa ctttcaccaa tggaccagaa ctacctgtga aattaataac agacatactc
     3781 caagctgcct ttgtgtgctt aatcacgtat actcacgtgc tcaatagtca ccaatgccct
     3841 ccctcttggc cctctccttt tcttttttcg accgaattaa ttcgtaatca tgtcatagct
     3901 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3961 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4021 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4081 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4141 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4201 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4261 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4321 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4381 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4441 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4501 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4561 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4621 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4681 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4741 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4801 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4861 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4921 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4981 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5041 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5101 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5161 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5221 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5281 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5341 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5401 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5461 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5521 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5581 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5641 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5701 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5761 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5821 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5881 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5941 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6001 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     6061 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     6121 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     6181 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     6241 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccataa
     6301 cgcatttaag cataaacacg cactatgccg ttcttctcat gtatatatat atacaggcaa
     6361 cacgcagata taggtgcgac gtgaacagtg agctgtatgt gcgcagctcg cgttgcattt
     6421 tcggaagcgc tcgttttcgg aaacgctttg aagttcctat tccgaagttc ctattctcta
     6481 gctagaaagt ataggaactt cagagcgctt ttgaaaacca aaagcgctct gaagacgcac
     6541 tttcaaaaaa ccaaaaacgc accggactgt aacgagctac taaaatattg cgaataccgc
     6601 ttccacaaac attgctcaaa agtatctctt tgctatatat ctctgtgcta tatccctata
     6661 taacctaccc atccaccttt cgctccttga acttgcatct aaactcgacc tctacatttt
     6721 ttatgtttat ctctagtatt actctttaga caaaaaaatt gtagtaagaa ctattcatag
     6781 agtgaatcga aaacaatacg aaaatgtaaa catttcctat acgtagtata tagagacaaa
     6841 atagaagaaa ccgttcataa ttttctgacc aatgaagaat catcaacgct atcactttct
     6901 gttcacaaag tatgcgcaat ccacatcggt atagaatata atcggggatg cctttatctt
     6961 gaaaaaatgc acccgcagct tcgctagtaa tcagtaaacg cgggaagtgg agtcaggctt
     7021 tttttatgga agagaaaata gacaccaaag tagccttctt ctaaccttaa cggacctaca
     7081 gtgcaaaaag ttatcaagag actgcattat agagcgcaca aaggagaaaa aaagtaatct
     7141 aagatgcttt gttagaaaaa tagcgctctc gggatgcatt tttgtagaac aaaaaagaag
     7201 tatagattct ttgttggtaa aatagcgctc tcgcgttgca tttctgttct gtaaaaatgc
     7261 agctcagatt ctttgtttga aaaattagcg ctctcgcgtt gcatttttgt tttacaaaaa
     7321 tgaagcacag attcttcgtt ggtaaaatag cgctttcgcg ttgcatttct gttctgtaaa
     7381 aatgcagctc agattctttg tttgaaaaat tagcgctctc gcgttgcatt tttgttctac
     7441 aaaatgaagc acagatgctt cgttgcttgc atgcaacttc ttttcttttt ttttcttttc
     7501 tctctccccc gttgttgtct caccatatcc gcaatgacaa aaaaatgatg gaagacacta
     7561 aa
//
