LOCUS       sequence-358268-        9182 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression GAL4-fusion of Co-NF-kB in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9182)
  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 9182)
  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-NF-kB
FEATURES             Location/Qualifiers
     source          1..9182
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        944..1345
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1373..1813
                     /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      5519..5706
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5574..5598)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(5759..6433)
                     /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(6434..6535)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(6549..6565)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6549..6565)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6562..6584)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6573..6589
                     /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(6597..6603,6604..6621,6622..6627))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6642..6663
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6780..6797)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6951..7539)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7031..7050)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7710..8501,8502..8570))
                     /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     8333..8352
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8571..8675)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8743..8761
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8799..8821)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8921..8940
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      join(8957..9182,1..939)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 gctctgaaga cgcactttca aaaaaccaaa aacgcaccgg actgtaacga gctactaaaa
       61 tattgcgaat accgcttcca caaacattgc tcaaaagtat ctctttgcta tatatctctg
      121 tgctatatcc ctatataacc tacccatcca cctttcgctc cttgaacttg catctaaact
      181 cgacctctac attttttatg tttatctcta gtattactct ttagacaaaa aaattgtagt
      241 aagaactatt catagagtga atcgaaaaca atacgaaaat gtaaacattt cctatacgta
      301 gtatatagag acaaaataga agaaaccgtt cataattttc tgaccaatga agaatcatca
      361 acgctatcac tttctgttca caaagtatgc gcaatccaca tcggtataga atataatcgg
      421 ggatgccttt atcttgaaaa aatgcacccg cagcttcgct agtaatcagt aaacgcggga
      481 agtggagtca ggcttttttt atggaagaga aaatagacac caaagtagcc ttcttctaac
      541 cttaacggac ctacagtgca aaaagttatc aagagactgc attatagagc gcacaaagga
      601 gaaaaaaagt aatctaagat gctttgttag aaaaatagcg ctctcgggat gcatttttgt
      661 agaacaaaaa agaagtatag attctttgtt ggtaaaatag cgctctcgcg ttgcatttct
      721 gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc gcgttgcatt
      781 tttgttttac aaaaatgaag cacagattct tcgttggtaa aatagcgctt tcgcgttgca
      841 tttctgttct gtaaaaatgc agctcagatt ctttgtttga aaaattagcg ctctcgcgtt
      901 gcatttttgt tctacaaaat gaagcacaga tgcttcgttg cttgcatgca acttcttttc
      961 tttttttttc ttttctctct cccccgttgt tgtctcacca tatccgcaat gacaaaaaaa
     1021 tgatggaaga cactaaagga aaaaattaac gacaaagaca gcaccaacag atgtcgttgt
     1081 tccagagctg atgaggggta tctcgaagca cacgaaactt tttccttcct tcattcacgc
     1141 acactactct ctaatgagca acggtatacg gccttccttc cagttacttg aatttgaaat
     1201 aaaaaaaagt ttgctgtctt gctatcaagt ataaatagac ctgcaattat taatcttttg
     1261 tttcctcgtc attgttctcg ttccctttct tccttgtttc tttttctgca caatatttca
     1321 agctatacca agcatacaat caactccaag cttgaagcaa gcctcctgaa agatgaagct
     1381 actgtcttct atcgaacaag catgcgatat ttgccgactt aaaaagctca agtgctccaa
     1441 agaaaaaccg aagtgcgcca agtgtctgaa gaacaactgg gagtgtcgct actctcccaa
     1501 aaccaaaagg tctccgctga ctagggcaca tctgacagaa gtggaatcaa ggctagaaag
     1561 actggaacag ctatttctac tgatttttcc tcgagaagac cttgacatga ttttgaaaat
     1621 ggattcttta caggatataa aagcattgtt aacaggatta tttgtacaag ataatgtgaa
     1681 taaagatgcc gtcacagata gattggcttc agtggagact gatatgcctc taacattgag
     1741 acagcataga ataagtgcga catcatcatc ggaagagagt agtaacaaag gtcaaagaca
     1801 gttgactgta tcgccggaat tcgacctttc tgaactttcc ggatgggacc cgaatttgtc
     1861 cttgcaggag cacactgcaa atctgctcgc tatggatgat tcaacaatgg cagctatgat
     1921 actgcattct gatggactgt cattgttcag agatatgggt ctgtacaatt ccgtctctac
     1981 gagcctggat attagcggta ttccaaaatt tccgccaccg ccacagcagc cgcaacttgc
     2041 acaagcgccg cctcggagag gtcacaacca atccagcagt agcgatagcc atagcacacc
     2101 atcacccggt tccgtgctct tctccccgag cccggcttct caagatatga gcctccagag
     2161 tccggagctg ggcctcgttg ggcttactgg caacagtagg ctggcgtcaa cgtcagaggc
     2221 cgatgatatt ctgcttgcga atatactggg gcacccagta cggtcagtta gtgcaaatac
     2281 atcaatggtt ggtcttccgg atgacctggg attctccccc acgaccggaa tggatctcac
     2341 aatcacggcg acgagtccag cgacggcaga ctcatccgcg tcagctacag ccttccctgc
     2401 ggcgtccccg atagcttcac ctagcgtctc caccagctcc gggcctgtaa ctgttggcgg
     2461 aaccagcgcg gccttggccg cattgactcc ggacagaatt aataggcttc ttagcgcagc
     2521 cgaggcggca gccgaagggg cggccaccct cgttgaggac cttcttatgg tcacggagga
     2581 gcccgcgcag ttcgcccgat tccgctacat gagcgagcag cgagagagat ctttggccgg
     2641 cgagaactcc ttccccacgc ttatggtaaa ccctaagtat gccagagtgg tccccgagat
     2701 ggccctcgta actgctgtcc tggtcacaaa aatgccggat ccccacactg ggaggcagca
     2761 gaagcattgg catcaccttg ggggcatacc tgcggctcct ctggaagggc ctcaacgcat
     2821 agcaaggttc gacaacattg ctgtaataat ggacaaggcg aacaataagg ataaggacaa
     2881 atcaaaggca cccgtaagat ctaaagacga tcagcgatgt gtcaggataa tgtttgaact
     2941 tgtctttgtg tcaggtaata cacagttcta tggcagggca atcagccaac caatctacaa
     3001 cgccaaattg gccatcacaa agattagtca tagcagcggg cccgtgaccg gtggtaacga
     3061 ggttatcatg ttgtgcagta agatcagaaa aggcgtgacc ggggtacgga tgaccgatcc
     3121 aacccaatgg tcagtccagg cacctagcgg ctctgcgtgg gagttgaatc cacaaacgct
     3181 gaaggccgac tgcaatgtac cgggcgctaa tctttttttc catcatcaat atgcagtcgt
     3241 cctgacgctg ccgccctacc acacgcagac cataacagcc cctgtcaccg tcaggatctc
     3301 aatcttggac actgacgatg agacagagag tcaatacgtt gagtatacct atttgccggc
     3361 cgaagcggcg gtgcgcaacg cagagctggc ggctcggaaa cggagacggg atgattcaat
     3421 gagagatttt atggaccgct ttgatgggtc cgacggtgga aatggctctg ggagtggccg
     3481 aggtaacaac ggaggacacg atggctctga cgctaacaat aacggccgag gcggtggcgg
     3541 tgggagttct tcaagtaaag gcggggacga accgtttaat tttaattcac tcattccaat
     3601 gcaccaacac aagcttcatc aactggcgct ctccacggta agggctgtgc aagggttcgc
     3661 ggccagtggg gatgcacggt atctgttggc cttgcataga cagttgcttg cagctccgaa
     3721 cgaaaacggt gactccccgc tgcacacagc agtagcacag ggtaatctca gatctactat
     3781 ggcattgctc ccattgcttg cggcagaaga tctccaaagt gtgaacgaca tgggggaaac
     3841 tgtcttgcat agcgctgtaa ttgagaaaag ggcagccata gctcggctgc ttctggtcgc
     3901 tggcgcagat cttggccaga gtaatgcacg gaactttaat cgaaacagcc ttcactacct
     3961 tgcccggcat ggagataggg cgactgcaat ggctgtgttc ggcgtttttg gcagtgcgca
     4021 agccccacca gctaacacga acacgcccgc acaggcaccg gcaggcgaaa cgaagccgaa
     4081 accagctgac ttgagactgt tggcaaggat tcaggcgcag gctataaaag cgctcctggc
     4141 ctgtgaactc gaaactgggg ccactcccgc ccatttggcg atacggggtg gccactggca
     4201 cgtctttgaa gcgtgtgcta agttggcggc atcagcaccc atacccaaag ccgctggttc
     4261 tcttctttct atggtcgctg aaaagagtag tggacatagt ctgttgcact cctgcgtcct
     4321 ggccaataat gagcaggctg ttcgacttct tataaatctg ggggcctctg gcaacgctag
     4381 ggacttcggg aagaacacgc cacttcacct ggccgcacga caaggacaca tcggcatagc
     4441 cgcgctgttg gtagaggcag gggctaccct gagtcttaac gccgttagcc agactcccct
     4501 cgatgttttg acttcagagg gttccggatt gagcagagat cagctccggg cattggtagc
     4561 agtcctccga ggcgagctca aatatgccga catgcgcggc cggccaaccc tccgaatgcc
     4621 gactcacgcg gagctgcata gcacggcggc cgcactcact tccgcctccc caggggcagt
     4681 ctcactggct gatttctacg cgggcaaaaa ggcctcaagg agccccgccc ctctcggtgc
     4741 ttcctcaagt cttcttagta gtactggagc ttctgcggct ggcgcctctg cgcccacgat
     4801 agcggcggtg catgccgcga gcgcaacacc ggtagagcgc accagcatga acaatgacga
     4861 tgattacgtt ctccttgaga aggacgcgcc ttaccctgtt gagcagcagc ctcacgggaa
     4921 gcgcaacaag cattctcacc acagattcac ccggtccagt cacggctcac aggataaaga
     4981 tgagcttaaa aaagataagg atgacccgaa aaaagaaaaa gagccgaagg aactgtcaaa
     5041 attcacactg aaagaggcgt ttgtggacgg tacgaatttt tgggaactta ctcggaaatt
     5101 cgctgggaag aagaaaatgg cttcagcaag tactggcgag atggagccgt tgagtccgga
     5161 acgaccactc agtccgacca atgcgggaag tggggctgcc tcacctttta accaagctaa
     5221 ggaacaagtt agccccggtg ctgtacctcc aacaggtctt gagaagcttg tcaacaagct
     5281 catggacgct tcagaggcta ccctctcaag tcaaccagct gaggccgtga cgcccgaaca
     5341 gaaactggct gaaaaacttg aaaaattggg actggcacct gcgtctacca ctagtgcgcc
     5401 gccaccgcac ccaaaagttg ctgctctgaa tgcgcagtct gtcgaagatg cgcggaagac
     5461 ttctacccat gccctttact ccgttgatta actcgacctg cagccaagct aattccgggc
     5521 gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa taagtgtata
     5581 caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga gtaactcttt
     5641 cctgtaggtc aggttgcttt ctcaggtata gcatgaggtc gctcttattg accacacctc
     5701 taccggcatg ccggcaagtg cacaaacaat acttaaataa atactactca gtaataacct
     5761 atttcttagc atttttgacg aaatttgcta ttttgttaga gtcttttaca ccatttgtct
     5821 ccacacctcc gcttacatca acaccaataa cgccatttaa tctaagcgca tcaccaacat
     5881 tttctggcgt cagtccacca gctaacataa aatgtaagct ttcggggctc tcttgccttc
     5941 caacccagtc agaaatcgag ttccaatcca aaagttcacc tgtcccacct gcttctgaat
     6001 caaacaaggg aataaacgaa tgaggtttct gtgaagctgc actgagtagt atgttgcagt
     6061 cttttggaaa tacgagtctt ttaataactg gcaaaccgag gaactcttgg tattcttgcc
     6121 acgactcatc tccatgcagt tggacgatat caatgccgta atcattgacc agagccaaaa
     6181 catcctcctt aggttgatta cgaaacacgc caaccaagta tttcggagtg cctgaactat
     6241 ttttatatgc ttttacaaga cttgaaattt tccttgcaat aaccgggtca attgttctct
     6301 ttctattggg cacacatata atacccagca agtcagcatc ggaatctaga gcacattctg
     6361 cggcctctgt gctctgcaag ccgcaaactt tcaccaatgg accagaacta cctgtgaaat
     6421 taataacaga catactccaa gctgcctttg tgtgcttaat cacgtatact cacgtgctca
     6481 atagtcacca atgccctccc tcttggccct ctccttttct tttttcgacc gaattaattc
     6541 gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6601 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6661 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6721 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6781 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6841 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6901 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6961 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     7021 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     7081 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     7141 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     7201 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7261 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7321 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7381 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7441 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7501 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7561 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7621 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7681 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7741 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7801 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7861 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7921 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7981 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     8041 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     8101 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     8161 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     8221 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     8281 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     8341 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     8401 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     8461 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     8521 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8581 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8641 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8701 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     8761 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     8821 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     8881 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     8941 ctgagagtgc accataacgc atttaagcat aaacacgcac tatgccgttc ttctcatgta
     9001 tatatatata caggcaacac gcagatatag gtgcgacgtg aacagtgagc tgtatgtgcg
     9061 cagctcgcgt tgcattttcg gaagcgctcg ttttcggaaa cgctttgaag ttcctattcc
     9121 gaagttccta ttctctagct agaaagtata ggaacttcag agcgcttttg aaaaccaaaa
     9181 gc
//
