LOCUS       sequence-356622-        6746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GAL4-fusion protein of Acanthoeca spectabilis NF-kB3 
            protein in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6746)
  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 6746)
  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-kB3
FEATURES             Location/Qualifiers
     source          1..6746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        225..626
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             654..1094
                     /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      2364..2551
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2419..2443)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2604..3278)
                     /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(3279..3380)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3394..3410)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3394..3410)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3407..3429)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3418..3434
                     /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(3442..3448,3449..3466,3467..3472))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3487..3508
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3625..3642)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3796..4384)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3876..3895)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4555..5346,5347..5415))
                     /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     5178..5197
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5416..5520)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5588..5606
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5644..5666)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5766..5785
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      join(5802..6746,1..220)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat
       61 ttttgtttta caaaaatgaa gcacagattc ttcgttggta aaatagcgct ttcgcgttgc
      121 atttctgttc tgtaaaaatg cagctcagat tctttgtttg aaaaattagc gctctcgcgt
      181 tgcatttttg ttctacaaaa tgaagcacag atgcttcgtt gcttgcatgc aacttctttt
      241 cttttttttt cttttctctc tcccccgttg ttgtctcacc atatccgcaa tgacaaaaaa
      301 atgatggaag acactaaagg aaaaaattaa cgacaaagac agcaccaaca gatgtcgttg
      361 ttccagagct gatgaggggt atctcgaagc acacgaaact ttttccttcc ttcattcacg
      421 cacactactc tctaatgagc aacggtatac ggccttcctt ccagttactt gaatttgaaa
      481 taaaaaaaag tttgctgtct tgctatcaag tataaataga cctgcaatta ttaatctttt
      541 gtttcctcgt cattgttctc gttccctttc ttccttgttt ctttttctgc acaatatttc
      601 aagctatacc aagcatacaa tcaactccaa gcttgaagca agcctcctga aagatgaagc
      661 tactgtcttc tatcgaacaa gcatgcgata tttgccgact taaaaagctc aagtgctcca
      721 aagaaaaacc gaagtgcgcc aagtgtctga agaacaactg ggagtgtcgc tactctccca
      781 aaaccaaaag gtctccgctg actagggcac atctgacaga agtggaatca aggctagaaa
      841 gactggaaca gctatttcta ctgatttttc ctcgagaaga ccttgacatg attttgaaaa
      901 tggattcttt acaggatata aaagcattgt taacaggatt atttgtacaa gataatgtga
      961 ataaagatgc cgtcacagat agattggctt cagtggagac tgatatgcct ctaacattga
     1021 gacagcatag aataagtgcg acatcatcat cggaagagag tagtaacaaa ggtcaaagac
     1081 agttgactgt atcgccggaa ttcccaaatg atttcgaggc tgttctgacg tctgggggtg
     1141 ggtctcatgc ttgcggacca tctgccgcca agagagctag ggtggacttg agtgatgcct
     1201 ccgaaaacgc ctataaccag gcatggagac cctccgcatc tgtggcttgt ggagctacct
     1261 ttcagccacc aaggacaccc atgactgcat gggacctggg cattcttcct cctacaactc
     1321 atacaggcca cggaggattc ccaccgcgag cataccccat tgagtctttc cctatgcatc
     1381 gaggtttcct tcaggttagt ggagctgcag cagccccgcc aataccgaga ttgatggggg
     1441 gccaccatca gtaccaccct acacacgcca tgtatcgcga tctgcgggaa gtcttggaaa
     1501 ttaccgagca gccgactgaa agagggcggt tcagatacgc aaaggaaaaa agacgaaccc
     1561 cactcccggg gagaagggat ggaggatttc ccacggttag ggttgctgac agatatcgag
     1621 acgtcttgcc agatggcact cacatacagg catccgtggt gacgagacag gacggagaag
     1681 acgggatccc acgccctcac tggcacagac tcgagggaag ggaaggagag agcgttagcc
     1741 agcctctcag cctgggagcc gctacattct ccaacctcgt ggtgatccgg agcgacagaa
     1801 atgccgcaga aaatcactgg ggccccaggc ctaccgaaga ccaacaggtg atccgaatta
     1861 tgttttccgt gcttttccgg ttgcctaccg gtgaaatggc ccggtcttgg gtggcttccg
     1921 aacctatcta cgggtgcgac ctgaaaattc agaacatgtc tcacacagaa gtcccgatgc
     1981 tctccggtgc tgaggtgacc ctgctgacta gtaagatcag gaaacagtcc atcgccctga
     2041 tgttggttga tgaatttaca gatcatgact tcgctcccgg aacaaagatt gacccagcaa
     2101 atgggtggga gtgtgatcag gacgggaaaa tctgttgcac tattcagccc agctacgtgc
     2161 accatcagta cgctctcgtg gcgcagatac cacagtattg ggacctgacc ttggagtctg
     2221 acagacaaat tagcgccagg attatcgata aggatcagaa gatggagtcc aactgttctc
     2281 aactcacata tgtccccgga tctgaggcca gaaggcgcag accaccaccc gaatgagtcg
     2341 acctgcagcc aagctaattc cgggcgaatt tcttatgatt tatgattttt attattaaat
     2401 aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa
     2461 aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagcatg
     2521 aggtcgctct tattgaccac acctctaccg gcatgccggc aagtgcacaa acaatactta
     2581 aataaatact actcagtaat aacctatttc ttagcatttt tgacgaaatt tgctattttg
     2641 ttagagtctt ttacaccatt tgtctccaca cctccgctta catcaacacc aataacgcca
     2701 tttaatctaa gcgcatcacc aacattttct ggcgtcagtc caccagctaa cataaaatgt
     2761 aagctttcgg ggctctcttg ccttccaacc cagtcagaaa tcgagttcca atccaaaagt
     2821 tcacctgtcc cacctgcttc tgaatcaaac aagggaataa acgaatgagg tttctgtgaa
     2881 gctgcactga gtagtatgtt gcagtctttt ggaaatacga gtcttttaat aactggcaaa
     2941 ccgaggaact cttggtattc ttgccacgac tcatctccat gcagttggac gatatcaatg
     3001 ccgtaatcat tgaccagagc caaaacatcc tccttaggtt gattacgaaa cacgccaacc
     3061 aagtatttcg gagtgcctga actattttta tatgctttta caagacttga aattttcctt
     3121 gcaataaccg ggtcaattgt tctctttcta ttgggcacac atataatacc cagcaagtca
     3181 gcatcggaat ctagagcaca ttctgcggcc tctgtgctct gcaagccgca aactttcacc
     3241 aatggaccag aactacctgt gaaattaata acagacatac tccaagctgc ctttgtgtgc
     3301 ttaatcacgt atactcacgt gctcaatagt caccaatgcc ctccctcttg gccctctcct
     3361 tttctttttt cgaccgaatt aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
     3421 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3481 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3541 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3601 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3661 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3721 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3781 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3841 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3901 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3961 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4021 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4081 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4141 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4201 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4261 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4321 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4381 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4441 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4501 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4561 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4621 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4681 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4741 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4801 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4861 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4921 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4981 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5041 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5101 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5161 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5221 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5281 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5341 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5401 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5461 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5521 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     5581 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     5641 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     5701 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     5761 ctatgcggca tcagagcaga ttgtactgag agtgcaccat aacgcattta agcataaaca
     5821 cgcactatgc cgttcttctc atgtatatat atatacaggc aacacgcaga tataggtgcg
     5881 acgtgaacag tgagctgtat gtgcgcagct cgcgttgcat tttcggaagc gctcgttttc
     5941 ggaaacgctt tgaagttcct attccgaagt tcctattctc tagctagaaa gtataggaac
     6001 ttcagagcgc ttttgaaaac caaaagcgct ctgaagacgc actttcaaaa aaccaaaaac
     6061 gcaccggact gtaacgagct actaaaatat tgcgaatacc gcttccacaa acattgctca
     6121 aaagtatctc tttgctatat atctctgtgc tatatcccta tataacctac ccatccacct
     6181 ttcgctcctt gaacttgcat ctaaactcga cctctacatt ttttatgttt atctctagta
     6241 ttactcttta gacaaaaaaa ttgtagtaag aactattcat agagtgaatc gaaaacaata
     6301 cgaaaatgta aacatttcct atacgtagta tatagagaca aaatagaaga aaccgttcat
     6361 aattttctga ccaatgaaga atcatcaacg ctatcacttt ctgttcacaa agtatgcgca
     6421 atccacatcg gtatagaata taatcgggga tgcctttatc ttgaaaaaat gcacccgcag
     6481 cttcgctagt aatcagtaaa cgcgggaagt ggagtcaggc tttttttatg gaagagaaaa
     6541 tagacaccaa agtagccttc ttctaacctt aacggaccta cagtgcaaaa agttatcaag
     6601 agactgcatt atagagcgca caaaggagaa aaaaagtaat ctaagatgct ttgttagaaa
     6661 aatagcgctc tcgggatgca tttttgtaga acaaaaaaga agtatagatt ctttgttggt
     6721 aaaatagcgc tctcgcgttg catttc
//
