LOCUS       sequence-358263-        7016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GAL4-fusion protein of Acanthoeca spectabilis NF-kB2 
            protein in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7016)
  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 7016)
  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-kB2
FEATURES             Location/Qualifiers
     source          1..7016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        89..490
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             518..958
                     /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      2498..2685
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2553..2577)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2738..3412)
                     /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(3413..3514)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3528..3544)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3528..3544)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3541..3563)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3552..3568
                     /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(3576..3582,3583..3600,3601..3606))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3621..3642
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3759..3776)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3930..4518)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4010..4029)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4689..5480,5481..5549))
                     /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     5312..5331
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5550..5654)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5722..5740
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5778..5800)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5900..5919
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      join(5936..7016,1..84)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 aatgcagctc agattctttg tttgaaaaat tagcgctctc gcgttgcatt tttgttctac
       61 aaaatgaagc acagatgctt cgttgcttgc atgcaacttc ttttcttttt ttttcttttc
      121 tctctccccc gttgttgtct caccatatcc gcaatgacaa aaaaatgatg gaagacacta
      181 aaggaaaaaa ttaacgacaa agacagcacc aacagatgtc gttgttccag agctgatgag
      241 gggtatctcg aagcacacga aactttttcc ttccttcatt cacgcacact actctctaat
      301 gagcaacggt atacggcctt ccttccagtt acttgaattt gaaataaaaa aaagtttgct
      361 gtcttgctat caagtataaa tagacctgca attattaatc ttttgtttcc tcgtcattgt
      421 tctcgttccc tttcttcctt gtttcttttt ctgcacaata tttcaagcta taccaagcat
      481 acaatcaact ccaagcttga agcaagcctc ctgaaagatg aagctactgt cttctatcga
      541 acaagcatgc gatatttgcc gacttaaaaa gctcaagtgc tccaaagaaa aaccgaagtg
      601 cgccaagtgt ctgaagaaca actgggagtg tcgctactct cccaaaacca aaaggtctcc
      661 gctgactagg gcacatctga cagaagtgga atcaaggcta gaaagactgg aacagctatt
      721 tctactgatt tttcctcgag aagaccttga catgattttg aaaatggatt ctttacagga
      781 tataaaagca ttgttaacag gattatttgt acaagataat gtgaataaag atgccgtcac
      841 agatagattg gcttcagtgg agactgatat gcctctaaca ttgagacagc atagaataag
      901 tgcgacatca tcatcggaag agagtagtaa caaaggtcaa agacagttga ctgtatcgcc
      961 ggaattcgag ccctggcgaa cacgccaata ctcacaaaaa cggtcaagaa ctgagagccc
     1021 gcccaccggc atcgccatgt ctccacataa agtgctcggc ttcggcgggg cagccgcagc
     1081 cgcagcagaa atggaagaag gtggcgctgc cgtaaccggc catgtgcctc tggggccgtt
     1141 cggaggtatg caatccctga caatgcacca cttcgacttg ccggacttgt ctgatcctag
     1201 gacgcctctg cggcttgatg acctcacgga aaaaccgcct ccaccgccgc acgatgccat
     1261 tcctccctgg cctaatacat tcggagctgg gaaatcccct gttggcccct tgccaccccc
     1321 tcataaacca acaaaggcag agcccatgtc tatcagtgaa ccgagttcac ctcgcaatga
     1381 cgcaagcaga gtcgtggaga tgtcagctga aggctatcca atagtatctg gggttcttag
     1441 aatcgtccag caacctgcgt ctcacgggcg gtttagatat tcaaaagaag gtaggaagac
     1501 gccgctccat ggacgggagg acggcagcta tcccacggta gcaatcgccg ataggtacag
     1561 gcacctggtg gaggaaggta cgcaagtaga tgtcaccctc gtaaccaaac ataacgacga
     1621 acatggatct cccattcaac actggcatgt tcttgaagga aaagaaggtg gcccagtctc
     1681 tcggcctctg aaggacggtg tcgcgacatt cccaaatctg gtggtaactc gcaccactgc
     1741 ggaaaaagag ggaggtgcgc gaaacgtaga ggatcagcat gttatcaggc tgatgtatac
     1801 aatgcggttt caggacgaga aaaggagaag cgtgctggcg agggcgataa gtgaacccat
     1861 ctttggtcaa gaggtcaaga tccaccgcat ttctcacatg cagattcctg ccacgggaaa
     1921 tatcgaagtc tttttcttga cctccaaaat taagcggaag aatactatat tgactttcac
     1981 tgagaccaca ccatcaagtt ttgatccggg gccacgatcc aagtggcagt tggacaaaaa
     2041 tatgtgtttg acatacgcca tgaaggattt gacggttcat tatcaatatg ctgtagtggc
     2101 tcgagtccct ccctattggg accagacgat cacctctccg cggaggatta aagtgtgcct
     2161 catagacacc gtccaaggcc tggaatcaaa tgtagttgaa attgattatg tccctgctcc
     2221 gttccgacca tactcttccg cgtctgctcc tcctggccac ccaccagtct tcggcatggc
     2281 accttctgtc catggatctt ggggagttcc tcaactcgat tggttcccgg cggataatgc
     2341 tgctggggcc gctgcagata tgcccatgag ggattccagt aattctccat ctgtaagcgc
     2401 gtcatccgaa tccacagata cacgaccttc tcgcgggagg gtatacgggt ccttcgagac
     2461 ttccgtttaa gtcgacctgc agccaagcta attccgggcg aatttcttat gatttatgat
     2521 ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta
     2581 ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc
     2641 tcaggtatag catgaggtcg ctcttattga ccacacctct accggcatgc cggcaagtgc
     2701 acaaacaata cttaaataaa tactactcag taataaccta tttcttagca tttttgacga
     2761 aatttgctat tttgttagag tcttttacac catttgtctc cacacctccg cttacatcaa
     2821 caccaataac gccatttaat ctaagcgcat caccaacatt ttctggcgtc agtccaccag
     2881 ctaacataaa atgtaagctt tcggggctct cttgccttcc aacccagtca gaaatcgagt
     2941 tccaatccaa aagttcacct gtcccacctg cttctgaatc aaacaaggga ataaacgaat
     3001 gaggtttctg tgaagctgca ctgagtagta tgttgcagtc ttttggaaat acgagtcttt
     3061 taataactgg caaaccgagg aactcttggt attcttgcca cgactcatct ccatgcagtt
     3121 ggacgatatc aatgccgtaa tcattgacca gagccaaaac atcctcctta ggttgattac
     3181 gaaacacgcc aaccaagtat ttcggagtgc ctgaactatt tttatatgct tttacaagac
     3241 ttgaaatttt ccttgcaata accgggtcaa ttgttctctt tctattgggc acacatataa
     3301 tacccagcaa gtcagcatcg gaatctagag cacattctgc ggcctctgtg ctctgcaagc
     3361 cgcaaacttt caccaatgga ccagaactac ctgtgaaatt aataacagac atactccaag
     3421 ctgcctttgt gtgcttaatc acgtatactc acgtgctcaa tagtcaccaa tgccctccct
     3481 cttggccctc tccttttctt ttttcgaccg aattaattcg taatcatgtc atagctgttt
     3541 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3601 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3661 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3721 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3781 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3841 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3901 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3961 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4021 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4081 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4141 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4201 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4261 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4321 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4381 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4441 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4501 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4561 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4621 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4681 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4741 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4801 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4861 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4921 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4981 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5041 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5101 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5161 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5221 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5281 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5341 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5401 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5461 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5521 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5581 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5641 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     5701 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     5761 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     5821 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     5881 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccataacgca
     5941 tttaagcata aacacgcact atgccgttct tctcatgtat atatatatac aggcaacacg
     6001 cagatatagg tgcgacgtga acagtgagct gtatgtgcgc agctcgcgtt gcattttcgg
     6061 aagcgctcgt tttcggaaac gctttgaagt tcctattccg aagttcctat tctctagcta
     6121 gaaagtatag gaacttcaga gcgcttttga aaaccaaaag cgctctgaag acgcactttc
     6181 aaaaaaccaa aaacgcaccg gactgtaacg agctactaaa atattgcgaa taccgcttcc
     6241 acaaacattg ctcaaaagta tctctttgct atatatctct gtgctatatc cctatataac
     6301 ctacccatcc acctttcgct ccttgaactt gcatctaaac tcgacctcta cattttttat
     6361 gtttatctct agtattactc tttagacaaa aaaattgtag taagaactat tcatagagtg
     6421 aatcgaaaac aatacgaaaa tgtaaacatt tcctatacgt agtatataga gacaaaatag
     6481 aagaaaccgt tcataatttt ctgaccaatg aagaatcatc aacgctatca ctttctgttc
     6541 acaaagtatg cgcaatccac atcggtatag aatataatcg gggatgcctt tatcttgaaa
     6601 aaatgcaccc gcagcttcgc tagtaatcag taaacgcggg aagtggagtc aggctttttt
     6661 tatggaagag aaaatagaca ccaaagtagc cttcttctaa ccttaacgga cctacagtgc
     6721 aaaaagttat caagagactg cattatagag cgcacaaagg agaaaaaaag taatctaaga
     6781 tgctttgtta gaaaaatagc gctctcggga tgcatttttg tagaacaaaa aagaagtata
     6841 gattctttgt tggtaaaata gcgctctcgc gttgcatttc tgttctgtaa aaatgcagct
     6901 cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat ttttgtttta caaaaatgaa
     6961 gcacagattc ttcgttggta aaatagcgct ttcgcgttgc atttctgttc tgtaaa
//
