LOCUS       sequence-436488-        6958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-tagged Acanthoeca spectabilis NF-kB2 protein in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6958)
  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 6958)
  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-FLAG-As-NF-kB2
FEATURES             Location/Qualifiers
     source          1..6958
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(103..897)
                     /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(214..233)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     824..843
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(964..1293)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      978..1113
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1032..1051)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1278..1298
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1307..1735)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1417..1438)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1629..1648
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    1781..2005
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1994..2011
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        2031..2049
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2031..2048
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(3633..3656)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3678..3697)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3679..3697)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3742..3945)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3771..3791)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3946..4325
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4497..4516
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4591..4695
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4696..4764,4765..5556)
                     /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(4914..4933)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5727..6315
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6216..6235
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6469..6486
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6603..6624
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6639..6644,6645..6662,6663..6669)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6677..6693
                     /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     6682..6704
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6701..6717
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6701..6717
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6766..6887
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6777..6796)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6831..6850
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt
       61 tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag
      121 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
      181 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
      241 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
      301 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc
      361 gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
      421 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
      481 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
      541 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
      601 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
      661 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag
      721 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
      781 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
      841 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg
      901 aaacgatcct catcctgtct cttgatcaga tccgaaaatg gatatacaag ctcccgggag
      961 ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga
     1021 ggcagaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga
     1081 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
     1141 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     1201 tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     1261 gcctggggac tttccacacc ctaactgaca cacattccac agaattaatt cgcgttaaat
     1321 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     1381 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     1441 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     1501 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     1561 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     1621 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     1681 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtgggga
     1741 taccccctag agccccagct ggttctttcc gcctcagaag ccatagagcc caccgcatcc
     1801 ccagcatgcc tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc caccccaccc
     1861 cccagaatag aatgacacct actcagacaa tgcgatgcaa tttcctcatt ttattaggaa
     1921 aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca
     1981 gatggctggc aactagaagg cacagtcgag gctgatcagc gagctctagc atttaggtga
     2041 cactatagaa tagggccctc tagatgcatg ctcgagcggc cgctcgaggg atccctgaat
     2101 tcttaaacgg aagtctcgaa ggacccgtat accctcccgc gagaaggtcg tgtatctgtg
     2161 gattcggatg acgcgcttac agatggagaa ttactggaat ccctcatggg catatctgca
     2221 gcggccccag cagcattatc cgccgggaac caatcgagtt gaggaactcc ccaagatcca
     2281 tggacagaag gtgccatgcc gaagactggt gggtggccag gaggagcaga cgcggaagag
     2341 tatggtcgga acggagcagg gacataatca atttcaacta catttgattc caggccttgg
     2401 acggtgtcta tgaggcacac tttaatcctc cgcggagagg tgatcgtctg gtcccaatag
     2461 ggagggactc gagccactac agcatattga taatgaaccg tcaaatcctt catggcgtat
     2521 gtcaaacaca tatttttgtc caactgccac ttggatcgtg gccccggatc aaaacttgat
     2581 ggtgtggtct cagtgaaagt caatatagta ttcttccgct taattttgga ggtcaagaaa
     2641 aagacttcga tatttcccgt ggcaggaatc tgcatgtgag aaatgcggtg gatcttgacc
     2701 tcttgaccaa agatgggttc acttatcgcc ctcgccagca cgcttctcct tttctcgtcc
     2761 tgaaaccgca ttgtatacat cagcctgata acatgctgat cctctacgtt tcgcgcacct
     2821 ccctcttttt ccgcagtggt gcgagttacc accagatttg ggaatgtcgc gacaccgtcc
     2881 ttcagaggcc gagagactgg gccaccttct tttccttcaa gaacatgcca gtgttgaatg
     2941 ggagatccat gttcgtcgtt atgtttggtt acgagggtga catctacttg cgtaccttcc
     3001 tccaccaggt gcctgtacct atcggcgatt gctaccgtgg gatagctgcc gtcctcccgt
     3061 ccatggagcg gcgtcttcct accttctttt gaatatctaa accgcccgtg agacgcaggt
     3121 tgctggacga ttctaagaac cccagatact attggatagc cttcagctga catctccacg
     3181 actctgcttg cgtcattgcg aggtgaactc ggttcactga tagacatggg ctctgccttt
     3241 gttggtttat gagggggtgg caaggggcca acaggggatt tcccagctcc gaatgtatta
     3301 ggccagggag gaatggcatc gtgcggcggt ggaggcggtt tttccgtgag gtcatcaagc
     3361 cgcagaggcg tcctaggatc agacaagtcc ggcaagtcga agtggtgcat tgtcagggat
     3421 tgcatacctc cgaacggccc cagaggcaca tggccggtta cggcagcgcc accttcttcc
     3481 atttctgctg cggctgcggc tgccccgccg aagccgagca ctttatgtgg agacatggcg
     3541 atgccggtgg gcgggctctc agttcttgac cgtttttgtg agtattggcg tgttcgccag
     3601 ggctcagaat tcgcatctgc agcacgtgtt aacttgtcat cgtcgtcctt gtagtccatg
     3661 gtaccaagct tgggtctccc tatagtgagt cgtattaatt tcgataagcc agtaagcagt
     3721 gggttctcta gttagccaga gagctctgct tatatagacc tcccaccgta cacgcctacc
     3781 gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg
     3841 gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc
     3901 aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg taatagcgat
     3961 gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata
     4021 atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata
     4081 cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg
     4141 gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg
     4201 gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat
     4261 atatgggcta tgaactaatg accccgtaat tgattactat taataactag tcaataatca
     4321 atgtcaacgc gtatatctgg cccgtacatc gcgaagcagc gcaaaacgcc taaccctaag
     4381 cagattcttc atgcaattgt cggtcaagcc ttgccttgtt gtagcttaaa ttttgctcgc
     4441 gcactactca gcgacctcca acacacaagc agggagcaga tactggctta actatgcggc
     4501 atcagagcag attgtactga gagtgcacca taggggatcg ggagatctcc cgatccgtcg
     4561 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4621 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4681 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4741 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4801 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4861 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4921 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4981 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5041 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5101 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5161 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5221 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5281 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5341 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5401 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5461 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5521 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5581 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5641 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5701 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5761 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5821 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5881 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5941 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6001 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6061 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6121 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6181 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6241 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6301 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6361 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6421 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     6481 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     6541 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     6601 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     6661 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     6721 gattacgcca agctctagct agaggtcgac ggtatacaga catgataaga tacattgatg
     6781 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     6841 atgctattgc tttatttgta accattataa gctgcaataa acaagttggg gtgggcgaag
     6901 aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacga
//
