LOCUS       sequence-444956-        8132 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MYC-tagged Acanthoeca spectabilis NF-kB3 protein in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8132)
  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 8132)
  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: pcDNA-MYC-As-NF-kB3
FEATURES             Location/Qualifiers
     source          1..8132
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2134..2163)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(2197..2226)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(2236..2265)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(2275..2304)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(2314..2343)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(2353..2382)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2437..2456)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2438..2456)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2501..2704)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2530..2550)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2705..3084
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3256..3275
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3350..3454
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3455..3523,3524..4315)
                     /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     complement(3673..3692)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4486..5074
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4975..4994
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5228..5245
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5362..5383
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5398..5403,5404..5421,5422..5428)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5436..5452
                     /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)."
     primer_bind     5441..5463
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5460..5476
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5460..5476
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5525..5646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5536..5555)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5590..5609
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5820..6614)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5931..5950)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6541..6560
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6681..7010)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6695..6830
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6749..6768)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6995..7015
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(7024..7452)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7134..7155)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7346..7365
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    7498..7722
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     7711..7728
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
ORIGIN
        1 tccatcacaa tcttgtgctg ctcggcctcc tccttcagct tatcgatcac ctcctgtttg
       61 gccaccaggg cctcctcggc ctgctggagc tgctgtttga gatcttccag ctgcattcct
      121 cgcttccgct cactgcccac cacgctgctc ttgatgtggt tgtcgtattc ttggaagagc
      181 tggtgatagg ccacctgcaa ctgggccagc ttcctcttct cctccgaggc ggcctggcgc
      241 tccatgcgga gcgcggcctc cacgctctgg ccctgcatgc gcagctggtc cacctgcacg
      301 ctgtgctgct gctgcagcgc ctcgcgctca ctctccagct gccgcgcctg ctcgctggcc
      361 gcccgggccc gaccctccag agcctggcat tccttagtgg cagcctccaa gcgactctgg
      421 ctctcctgca gctccccgag caaggacgtc acctgggctt tcacagaggc cttgtcctca
      481 gccatctgct gctggcatct cttcaggtgc tccacctccc gcagagcctg ctccttctgc
      541 ctcttcagat cgagcttctc caggccgagt ctctccacca gtttcctggc ctcctggaac
      601 ttgcacatga ggaactcctt ctcctccctc tggctggctt ggaaatgcag aagctcctcg
      661 cagcgctccc gcagaatctg gttgctctgc cggatggcat ctcggagctc ttgattctcc
      721 tccaggcagc gctggagggt ctcaggagcg ccctgttctg aaggcaggtg cagcatggct
      781 ggcttcccca gaggagactc ttcgcccagt acgtcctgat ctgctgccgg gccaccactg
      841 ggctgcacca tctcacacag ttggctcttc cagaggtgcc tattcatgga attctcattc
      901 gggtggtggt ctgcgccttc tggcctcaga tccggggaca tatgtgagtt gagaacagtt
      961 ggactccatc ttctgatcct tatcgataat cctggcgcta atttgtctgt cagactccaa
     1021 ggtcaggtcc caatactgtg gtatctgcgc cacgagagcg tactgatggt gcacgtagct
     1081 gggctgaata gtgcaacaga ttttcccgtc ctgatcacac tcccacccat ttgctgggtc
     1141 aatctttgtt ccgggagcga agtcatgatc tgtaaattca tcaaccaaca tcagggcgat
     1201 ggactgtttc ctgatcttac tagtcagcag ggtcacctca gcaccggaga gcatcgggac
     1261 ttctgtgtga gacatgttct gaattttcag gtcgcacccg tagataggtt cggaagccac
     1321 ccaagaccgg gccatttcac cggtaggcaa ccggaaaagc acggaaaaca taattcggat
     1381 cacctgttgg tcttcggtag gcctggggcc ccagtgattt tctgcggcat ttctgtcgct
     1441 ccggatcacc acgaggttgg agaatgtagc ggctcccagg ctgagaggct ggctaacgct
     1501 ctctccttcc cttccctcga gtctgtgcca gtgagggcgt gggatcccgt cttctccgtc
     1561 ctgtctcgtc accacggatg cctgtatgtg agtgccatct ggcaagacgt ctcgatatct
     1621 gtcagcaacc ctaaccgtgg gaaatcctcc atcccttctc cccgggagtg gggttcgtct
     1681 tttttccttt gcgtatctga accgccctct ttcagtcggc tgctcggtaa tttccaagac
     1741 ttcccgcaga tcgcgataca tggcgtgtgt agggtggtac tgatggtggc cccccatcaa
     1801 tctcggtatt ggcggggctg ctgcagctcc actaacctga aggaaacctc gatgcatagg
     1861 gaaagactca atggggtatg ctcgcggtgg gaatcctccg tggcctgtat gagttgtagg
     1921 aggaagaatg cccaggtccc atgcagtcat gggtgtcctt ggtggctgaa aggtagctcc
     1981 acaagccaca gatgcggagg gtctccatgc ctggttatag gcgttttcgg aggcatcact
     2041 caagtccacc ctagctctct tggcggcaga tggtccgcaa gcatgagacc cacccccaga
     2101 cgtcagaaca gcctcgaaat catttggaga attcaagtcc tcttcagaaa tgagcttttg
     2161 ctccatggtg aggtcgccca agctctccat ttcattcaag tcctcttcag aaatgagctt
     2221 ttgctccatt tcattcaagt cctcttcaga aatgagcttt tgctccattt cattcaagtc
     2281 ctcttcagaa atgagctttt gctccatttc attcaagtcc tcttcagaaa tgagcttttg
     2341 ctccatttca ttcaagtcct cttcagaaat gagcttttgc tccatagctt taaatcggta
     2401 ccaagcttaa gtttaaacgc tagccagctt gggtctccct atagtgagtc gtattaattt
     2461 cgataagcca gtaagcagtg ggttctctag ttagccagag agctctgctt atatagacct
     2521 cccaccgtac acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg
     2581 gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac
     2641 ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca
     2701 tcaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata
     2761 aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg
     2821 gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact
     2881 ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta
     2941 ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt
     3001 atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt
     3061 aataactagt caataatcaa tgtcaacgcg tatatctggc ccgtacatcg cgaagcagcg
     3121 caaaacgcct aaccctaagc agattcttca tgcaattgtc ggtcaagcct tgccttgttg
     3181 tagcttaaat tttgctcgcg cactactcag cgacctccaa cacacaagca gggagcagat
     3241 actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat aggggatcgg
     3301 gagatctccc gatccgtcga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3361 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3421 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3481 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3541 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3601 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3661 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3721 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3781 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3841 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3901 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3961 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4021 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4081 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4141 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4201 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4261 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4321 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4381 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4441 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4501 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4561 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4621 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4681 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4741 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4801 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4861 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4921 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4981 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5041 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5101 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5161 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     5221 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     5281 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     5341 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     5401 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     5461 aggaaacagc tatgaccatg attacgccaa gctctagcta gaggtcgacg gtatacagac
     5521 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     5581 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     5641 caagttgggg tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc
     5701 ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa
     5761 atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct
     5821 cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata
     5881 ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg
     5941 gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat
     6001 ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg
     6061 acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg
     6121 agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta
     6181 cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc
     6241 gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga
     6301 gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca
     6361 gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc
     6421 gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc
     6481 gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt
     6541 gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca
     6601 tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcagatcc gaaaatggat
     6661 atacaagctc ccgggagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac
     6721 ttctggaata gctcagaggc agaggcggcc tcggcctctg cataaataaa aaaaattagt
     6781 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt
     6841 aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct
     6901 ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata
     6961 cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac attccacaga
     7021 attaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     7081 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     7141 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     7201 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     7261 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     7321 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     7381 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     7441 tacagggcgc gtggggatac cccctagagc cccagctggt tctttccgcc tcagaagcca
     7501 tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt
     7561 cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt
     7621 cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg
     7681 gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct gatcagcggg
     7741 tttaaacggg ccctctagac tactcaatgc actccatgac atgtatctgc agggtgtcca
     7801 tatcaggggc ctgatactgg cacttgggac agcagaagtc aggtggctcc tcgggggggc
     7861 tcctcctctg gctgggcagg gccaggggag aggagaggta ggcaggggcg gggggcaagg
     7921 gggcctggga gacctcgaca tgccgcttcc tcatgtcctc gatcctggcc gactcctgac
     7981 agctggcctt cagtttgctg tactccctct gcagctgctc cagctgctcc tgcaggagct
     8041 ccttcttctc ggccagcttc tcccgggcct gcctctcagc ctggaagtcc gccttgtaga
     8101 tatccgcctg ggccttcagc accggaacgg tc
//
