LOCUS       sequence-444950-        7175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-tagged version of the Co-NF-kB Rel Homology Domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7175)
  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 7175)
  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-Co-RHD
FEATURES             Location/Qualifiers
     source          1..7175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(162..181)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        353..732
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        733..936
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     887..907
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     981..1000
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        981..999
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1022..1045
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2846..2864)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2847..2864)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2884..2901)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2890..3114
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3160..3588
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3247..3266)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3457..3478
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3597..3617)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3602..3931
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3782..3917
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3844..3863
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3998..4792
                     /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(4052..4071)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4662..4681
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4966..5087
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5003..5022)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5057..5076
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5136..5152)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5136..5152)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5149..5171)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5160..5176
                     /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(5184..5190,5191..5208,5209..5214))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5229..5250
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5367..5384)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5538..6126)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5618..5637)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6297..7088,7089..7157))
                     /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     6920..6939
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7158..7175,1..87))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
       61 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
      121 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
      181 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
      241 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
      301 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
      361 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      421 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      481 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      541 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      601 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      661 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      721 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      781 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      841 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      901 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
      961 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc ttggtaccat
     1021 ggactacaag gacgacgatg acaagttaac acgtgctgca gatgcgaatt cagacctttc
     1081 tgaactttcc ggatgggacc cgaatttgtc cttgcaggag cacactgcaa atctgctcgc
     1141 tatggatgat tcaacaatgg cagctatgat actgcattct gatggactgt cattgttcag
     1201 agatatgggt ctgtacaatt ccgtctctac gagcctggat attagcggta ttccaaaatt
     1261 tccgccaccg ccacagcagc cgcaacttgc acaagcgccg cctcggagag gtcacaacca
     1321 atccagcagt agcgatagcc atagcacacc atcacccggt tccgtgctct tctccccgag
     1381 cccggcttct caagatatga gcctccagag tccggagctg ggcctcgttg ggcttactgg
     1441 caacagtagg ctggcgtcaa cgtcagaggc cgatgatatt ctgcttgcga atatactggg
     1501 gcacccagta cggtcagtta gtgcaaatac atcaatggtt ggtcttccgg atgacctggg
     1561 attctccccc acgaccggaa tggatctcac aatcacggcg acgagtccag cgacggcaga
     1621 ctcatccgcg tcagctacag ccttccctgc ggcgtccccg atagcttcac ctagcgtctc
     1681 caccagctcc gggcctgtaa ctgttggcgg aaccagcgcg gccttggccg cattgactcc
     1741 ggacagaatt aataggcttc ttagcgcagc cgaggcggca gccgaagggg cggccaccct
     1801 cgttgaggac cttcttatgg tcacggagga gcccgcgcag ttcgcccgat tccgctacat
     1861 gagcgagcag cgagagagat ctttggccgg cgagaactcc ttccccacgc ttatggtaaa
     1921 ccctaagtat gccagagtgg tccccgagat ggccctcgta actgctgtcc tggtcacaaa
     1981 aatgccggat ccccacactg ggaggcagca gaagcattgg catcaccttg ggggcatacc
     2041 tgcggctcct ctggaagggc ctcaacgcat agcaaggttc gacaacattg ctgtaataat
     2101 ggacaaggcg aacaataagg ataaggacaa atcaaaggca cccgtaagat ctaaagacga
     2161 tcagcgatgt gtcaggataa tgtttgaact tgtctttgtg tcaggtaata cacagttcta
     2221 tggcagggca atcagccaac caatctacaa cgccaaattg gccatcacaa agattagtca
     2281 tagcagcggg cccgtgaccg gtggtaacga ggttatcatg ttgtgcagta agatcagaaa
     2341 aggcgtgacc ggggtacgga tgaccgatcc aacccaatgg tcagtccagg cacctagcgg
     2401 ctctgcgtgg gagttgaatc cacaaacgct gaaggccgac tgcaatgtac cgggcgctaa
     2461 tctttttttc catcatcaat atgcagtcgt cctgacgctg ccgccctacc acacgcagac
     2521 cataacagcc cctgtcaccg tcaggatctc aatcttggac actgacgatg agacagagag
     2581 tcaatacgtt gagtatacct atttgccggc cgaagcggcg gtgcgcaacg cagagctggc
     2641 ggctcggaaa cggagacggg atgattcaat gagagatttt atggaccgct ttgatgggtc
     2701 cgacggtgga aatggctctg ggagtggccg aggtaacaac ggaggacacg atggctctga
     2761 cgctaacaat aacggccgag gcggtggcgg tgggagttct tcaagtaaag gcgggtaact
     2821 cgagcatgca tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat
     2881 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2941 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3001 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3061 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     3121 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3181 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3241 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3301 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3361 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3421 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3481 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3541 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     3601 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3661 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3721 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3781 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3841 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     3901 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     3961 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4021 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4081 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4141 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4201 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4261 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4321 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4381 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4441 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4501 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4561 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4621 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4681 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4741 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4801 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4861 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4921 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4981 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5041 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5101 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5161 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5221 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5281 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5341 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5401 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5461 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5521 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5581 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5641 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5701 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5761 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5821 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5881 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5941 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6001 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6061 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6121 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6181 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6241 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6301 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6361 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6421 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6481 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6541 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6601 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6661 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6721 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6781 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6841 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6901 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6961 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7021 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7081 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7141 aaatgttgaa tactcatact cttccttttt caata
//
