LOCUS       sequence-364072-        6737 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GAL4-fusion protein of Acanthoeca spectabilis NF-kB1 
            protein in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6737)
  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 6737)
  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-As-NF-kB1
FEATURES             Location/Qualifiers
     source          1..6737
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        42..443
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             471..911
                     /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"
     regulatory      1074..1083
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      2172..2359
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2227..2251)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2412..3086)
                     /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(3087..3188)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3202..3218)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3202..3218)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3215..3237)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3226..3242
                     /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(3250..3256,3257..3274,3275..3280))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3295..3316
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3433..3450)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3604..4192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3684..3703)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4363..5154,5155..5223))
                     /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     4986..5005
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5224..5328)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5396..5414
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5452..5474)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5574..5593
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      join(5610..6737,1..37)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 atttttgttc tacaaaatga agcacagatg cttcgttgct tgcatgcaac ttcttttctt
       61 tttttttctt ttctctctcc cccgttgttg tctcaccata tccgcaatga caaaaaaatg
      121 atggaagaca ctaaaggaaa aaattaacga caaagacagc accaacagat gtcgttgttc
      181 cagagctgat gaggggtatc tcgaagcaca cgaaactttt tccttccttc attcacgcac
      241 actactctct aatgagcaac ggtatacggc cttccttcca gttacttgaa tttgaaataa
      301 aaaaaagttt gctgtcttgc tatcaagtat aaatagacct gcaattatta atcttttgtt
      361 tcctcgtcat tgttctcgtt ccctttcttc cttgtttctt tttctgcaca atatttcaag
      421 ctataccaag catacaatca actccaagct tgaagcaagc ctcctgaaag atgaagctac
      481 tgtcttctat cgaacaagca tgcgatattt gccgacttaa aaagctcaag tgctccaaag
      541 aaaaaccgaa gtgcgccaag tgtctgaaga acaactggga gtgtcgctac tctcccaaaa
      601 ccaaaaggtc tccgctgact agggcacatc tgacagaagt ggaatcaagg ctagaaagac
      661 tggaacagct atttctactg atttttcctc gagaagacct tgacatgatt ttgaaaatgg
      721 attctttaca ggatataaaa gcattgttaa caggattatt tgtacaagat aatgtgaata
      781 aagatgccgt cacagataga ttggcttcag tggagactga tatgcctcta acattgagac
      841 agcatagaat aagtgcgaca tcatcatcgg aagagagtag taacaaaggt caaagacagt
      901 tgactgtatc gccggaattc gaagccagat ggcggcgggc ccctggcctg ggcaaccctc
      961 ggcctagcaa gcggtgcaga agccagagcc ctagccaaag acaggtgatc gccgagtctt
     1021 tcggcaaggg ccaggccgct gccgaagagg acgccagagg cgaggccggc cccgccgcca
     1081 tggtccctct gggaggcgcc atcgacttca ccgacgttgg catggccgct tctagctttg
     1141 accccgccgt gtggcacggc atggtggcca agagcaccat gctggcccaa cagggccaca
     1201 gaggaatttc tctgaacgac atcgcagtgg gcggattcga cgacaccgtg tccgaggtgg
     1261 acggcgtgct ggaaatcagc cagcagcctg ccgcccacgg aagattcaga tacagcaagg
     1321 agggcagaaa gacacctctg cctggagcta tcgagggcag cttccccacc gtgcagctga
     1381 gccccgctta tagacacatg gtgcccgacg gcaccatggt ttctgtgacc ctgatcacca
     1441 agtaccacga tgatagaggc gctccggtta gacactggca caccctcgag gtgaaggaag
     1501 gcggcgccgc ctctagacct ctgagcatgg gcctggtgca gttccctaat ctggtcgtga
     1561 ccagagccgc tatgcctgct gatctggacg atgctggagc tgcaagaaac ccggaagatc
     1621 agcacgtgat tagactgatg tttacaatga tgttccggga cagcgccgga gtgatcttca
     1681 agaccagcgt gatcagcaat cctatctacg gcatcgagct gaagatccac aagtccagcc
     1741 accccagagt gcctgtggcc ggccagctcg acgtcttttt cctgacatct aaggtgaaaa
     1801 agaaaaacac cctcatcaag gtgagagagg tgtaccccgc tcctttcgat cctggcccaa
     1861 acagcggctg ggagctggac gagagaggca gactgacctt caccgtggac aacctgcacg
     1921 tgcactacca gtacgccgtg gtcgccacag tgccacctta ctgggaccag acaatcacaa
     1981 caagcagagt gctggacatc tgcctggtgg ataccgccca gggactggaa agcaactgcg
     2041 tgcagatcga gtactgcccc cccagcgccg cagaaggcga aggcgctgct gccgccgctt
     2101 ctggcgccgc tagtgatgcc gcccctggcc ctgccctgat gtgagtcgac ctgcagccaa
     2161 gctaattccg ggcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa
     2221 aaataagtgt atacaaattt taaagtgact cttaggtttt aaaacgaaaa ttcttattct
     2281 tgagtaactc tttcctgtag gtcaggttgc tttctcaggt atagcatgag gtcgctctta
     2341 ttgaccacac ctctaccggc atgccggcaa gtgcacaaac aatacttaaa taaatactac
     2401 tcagtaataa cctatttctt agcatttttg acgaaatttg ctattttgtt agagtctttt
     2461 acaccatttg tctccacacc tccgcttaca tcaacaccaa taacgccatt taatctaagc
     2521 gcatcaccaa cattttctgg cgtcagtcca ccagctaaca taaaatgtaa gctttcgggg
     2581 ctctcttgcc ttccaaccca gtcagaaatc gagttccaat ccaaaagttc acctgtccca
     2641 cctgcttctg aatcaaacaa gggaataaac gaatgaggtt tctgtgaagc tgcactgagt
     2701 agtatgttgc agtcttttgg aaatacgagt cttttaataa ctggcaaacc gaggaactct
     2761 tggtattctt gccacgactc atctccatgc agttggacga tatcaatgcc gtaatcattg
     2821 accagagcca aaacatcctc cttaggttga ttacgaaaca cgccaaccaa gtatttcgga
     2881 gtgcctgaac tatttttata tgcttttaca agacttgaaa ttttccttgc aataaccggg
     2941 tcaattgttc tctttctatt gggcacacat ataataccca gcaagtcagc atcggaatct
     3001 agagcacatt ctgcggcctc tgtgctctgc aagccgcaaa ctttcaccaa tggaccagaa
     3061 ctacctgtga aattaataac agacatactc caagctgcct ttgtgtgctt aatcacgtat
     3121 actcacgtgc tcaatagtca ccaatgccct ccctcttggc cctctccttt tcttttttcg
     3181 accgaattaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac
     3241 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3301 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3361 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3421 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3481 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3541 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3601 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3661 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3721 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3781 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3841 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3901 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3961 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4021 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4081 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4141 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4201 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4261 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4321 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4381 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4441 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4501 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4561 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4621 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4681 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4741 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4801 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4861 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4921 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4981 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5041 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5101 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5161 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5221 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5281 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5341 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     5401 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     5461 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     5521 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     5581 agagcagatt gtactgagag tgcaccataa cgcatttaag cataaacacg cactatgccg
     5641 ttcttctcat gtatatatat atacaggcaa cacgcagata taggtgcgac gtgaacagtg
     5701 agctgtatgt gcgcagctcg cgttgcattt tcggaagcgc tcgttttcgg aaacgctttg
     5761 aagttcctat tccgaagttc ctattctcta gctagaaagt ataggaactt cagagcgctt
     5821 ttgaaaacca aaagcgctct gaagacgcac tttcaaaaaa ccaaaaacgc accggactgt
     5881 aacgagctac taaaatattg cgaataccgc ttccacaaac attgctcaaa agtatctctt
     5941 tgctatatat ctctgtgcta tatccctata taacctaccc atccaccttt cgctccttga
     6001 acttgcatct aaactcgacc tctacatttt ttatgtttat ctctagtatt actctttaga
     6061 caaaaaaatt gtagtaagaa ctattcatag agtgaatcga aaacaatacg aaaatgtaaa
     6121 catttcctat acgtagtata tagagacaaa atagaagaaa ccgttcataa ttttctgacc
     6181 aatgaagaat catcaacgct atcactttct gttcacaaag tatgcgcaat ccacatcggt
     6241 atagaatata atcggggatg cctttatctt gaaaaaatgc acccgcagct tcgctagtaa
     6301 tcagtaaacg cgggaagtgg agtcaggctt tttttatgga agagaaaata gacaccaaag
     6361 tagccttctt ctaaccttaa cggacctaca gtgcaaaaag ttatcaagag actgcattat
     6421 agagcgcaca aaggagaaaa aaagtaatct aagatgcttt gttagaaaaa tagcgctctc
     6481 gggatgcatt tttgtagaac aaaaaagaag tatagattct ttgttggtaa aatagcgctc
     6541 tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga aaaattagcg
     6601 ctctcgcgtt gcatttttgt tttacaaaaa tgaagcacag attcttcgtt ggtaaaatag
     6661 cgctttcgcg ttgcatttct gttctgtaaa aatgcagctc agattctttg tttgaaaaat
     6721 tagcgctctc gcgttgc
//
