LOCUS       sequence-477299-        7212 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Kaede driven by NFkB activity can be switched from green to red 
            fluorescence by UV light exposure, enabling tracking of NFkB 
            positive cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7212)
  AUTHORS   Campbell CA, Calderon R, Pavani G, Cheng X, Barakat R, Snella E, Liu
            F, Peng X, Essner JJ, Dorman KS, McGrail M, Gadue P, French DL, 
            Espin-Palazon R
  TITLE     p65 signaling dynamics drive the developmental progression of 
            hematopoietic stem and progenitor cells through cell cycle 
            regulation.
  JOURNAL   Nat Commun. 2024 Sep 6;15(1):7787. doi: 10.1038/s41467-024-51922-5.
  PUBMED    39242546
REFERENCE   2  (bases 1 to 7212)
  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: "Nat 
            Commun."; date: "2024-09-6"; volume: "15(1):7787. doi"; pages: " 
            10.1038/s41467-024-51922-5"
COMMENT     Sequence Label: 6xHsa.NFKB:Kaede
FEATURES             Location/Qualifiers
     source          1..7212
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(32..43,44..206)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     protein_bind    585..609
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(747..771)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(937..948)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             1014..1691
                     /codon_start=1
                     /product="Kaede green-to-red photoconvertible fluorescent 
                     protein (Ando et al., 2002)"
                     /label=Kaede
                     /translation="MSLIKPEMKIKLLMEGNVNGHQFVIEGDGKGHPFEGKQSMDLVVK
                     EGAPLPFAYDILTTAFHYGNRVFAKYPDHIPDYFKQSFPKGFSWERSLMFEDGGVCIAT
                     NDITLKGDTFFNKVRFDGVNFPPNGPVMQKKTLKWEASTEKMYLRDGVLTGDITMALLL
                     KGDVHYRCDFRTTYKSRQEGVKLPGYHFVDHCISILRHDKDYNEVKLYEHAVAHSGLPD
                     NVK"
     polyA_signal    1701..1835
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1725..1744)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1779..1798
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(2274..2461,2462..2473)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2507..2523)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2507..2523)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2560..2580)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2560..2578)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2599..2615)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2599..2615)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2612..2634)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2623..2639
                     /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(2647..2653,2654..2671,2672..2677))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2692..2713
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2830..2847)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3001..3589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3081..3100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3760..4551,4552..4620))
                     /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     4383..4402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4621..4725)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4751..5206)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4888..4909)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5100..5119
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5333..5355
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5347..5364
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5348..5364
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5374..5393
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5374..5392
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 acaggtcaga ggtttgtcca ggagttcttg acagaggtgt aaaaagtact caaaaatttt
       61 actcaagtga aagtacaagt acttagggaa aattttactc aattaaaagt aaaagtatct
      121 ggctagaatc ttacttgagt aaaagtaaaa aagtactcca ttaaaattgt acttgagtat
      181 taaggaagta aaagtaaaag caagaaagaa aactagagat tcttgtttaa gcttttaatc
      241 tcaaaaaaca ttaaatgaaa tgcatacaag gttttatcct gctttagaac tgtttgtatt
      301 taattatcaa actataagac agacaatcta atgccagtac acgctactca aagttgtaaa
      361 acctcagatt taacttcagt agaagctgat tctcaaaatt gttagtgtca agcctagctc
      421 ttttggggct gaaaagcaat cctgcagtgc tgaaaagcct ctcacaggca gccgatgcgg
      481 gaagaggtgt attagtcttg atagagaggc tgcaaatagc aggaaacgtg agcagagact
      541 ccctggtgtc tgaaacacag gccagatggg ccctcgagat atcaacaagt ttgtacaaaa
      601 aagcaggctt cctgtgaatg catgcacggt cgagggactt tccgctgggg actttccgtc
      661 gagggacttt ccgctgggga ctttccgtcg agggactttc cgctggggac tttccgtcga
      721 cctgcagaat tcgaagcttg agctcgaccc agctttcttg tacaaagtgg ttgatatcca
      781 gtgacgtagg aagtccatcc attcacagcg cttctataaa ggcgccagct gaggcgccta
      841 ctactccaac cgcgactgca gcgagcaact gagaagactg gatagagccg gcggttccgc
      901 gaacgagcag tgaccgcgct cccacccagc tctgctctgc agctcccacc agtgtctacc
      961 cctggacccc ttgccgggct ttccccaaac ttcgaggatc caccggtcgc cacatgagtc
     1021 tgattaaacc agaaatgaag atcaagctgc ttatggaagg caatgtaaac gggcaccagt
     1081 ttgttattga gggagatgga aaaggccatc cttttgaggg aaaacagagt atggaccttg
     1141 tagtcaaaga aggcgcacct ctcccttttg cctacgatat cttgacaaca gcattccatt
     1201 atggtaacag ggtttttgct aaatacccag accatatacc agactacttc aagcagtcgt
     1261 ttcccaaagg gttttcttgg gagcgaagcc tgatgttcga ggacgggggc gtttgcatcg
     1321 ctacaaatga cataacactg aaaggagaca ctttttttaa caaagttcga tttgatggcg
     1381 taaactttcc cccaaatggt cctgttatgc agaagaagac tctgaaatgg gaggcatcca
     1441 ctgagaaaat gtatttgcgt gatggagtgt tgacgggcga tattaccatg gctctgctgc
     1501 ttaaaggaga tgtccattac cgatgtgact tcagaactac ttacaaatct aggcaggagg
     1561 gtgtcaagtt gccaggatat cactttgtcg atcactgcat cagcatattg aggcatgaca
     1621 aagactacaa cgaggttaag ctgtatgagc atgctgttgc ccattctgga ttgccggaca
     1681 acgtcaagta aatcgatgat gatccagaca tgataagata cattgatgag tttggacaaa
     1741 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     1801 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     1861 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     1921 gtggtatggc tgattatgat cctctagatc agatctgcga agatacggcc acgggtgctc
     1981 ttgatcctgt ggctgatttt ggactgtgct gctcgcagct gctgatgaat cacatacttc
     2041 ctccattttc ttccactgat tgactgttat aatttcccta atttccaggt caaggtgctg
     2101 tgcattgtgg taatagatgt gacatgacgt cacttccaaa ggaccaatga acatgtctga
     2161 ccaatttcat ataatgtgaa aacgattttc ataggcagaa taaataacat ttaaattaaa
     2221 ctgggcatca gcgcaattca attggtttgg taatagcaag ggaaaataga atgaagtgat
     2281 ctccaaaaaa taagtacttt ttgactgtaa ataaaattgt aaggagtaaa aagtactttt
     2341 ttttctaaaa aaatgtaatt aagtaaaagt aaaagtattg atttttaatt gtactcaagt
     2401 aaagtaaaaa tccccaaaaa taatacttaa gtacagtaat caagtaaaat tactcaagta
     2461 ctttacacct ctggttcttg accccctacc ttcagcaagc ccagcagatc cactagttct
     2521 agagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag ggttaattgc
     2581 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2641 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     2701 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2761 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2821 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2881 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2941 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3001 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3061 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3661 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3721 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3781 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3841 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3901 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3961 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4021 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4081 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4141 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4201 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4261 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4321 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4381 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4441 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4501 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4561 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4621 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4681 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4741 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     4801 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     4861 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     4921 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     4981 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     5041 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     5101 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     5161 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
     5221 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     5281 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     5341 cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg
     5401 ggtaccaaat agtaggaatt acccacctgt acaagtgctg aaaacttgga tgaataagcc
     5461 cgtttgccat tttctactgc tattttaaat cttttctgta tttgtctgca tttgtcttta
     5521 cccttgaata aatgttttag ttgttttttt tcaattatga ctgtgtttaa cagacattat
     5581 tacaataata tgtaatagta gtacacacta ttattatgta atagtacttg ttgactgtat
     5641 ttgagaactg gaccgagtga gtgttacgtc acccattcaa aatgacttac ttctggctcc
     5701 aacaaaatga agttaattca gttgccattt ttcactgtat ggacatcgcc gtgttggagc
     5761 tagacgttgc caataagcaa gaaagagccg agatgcgtcg agtctgagtc accgttccta
     5821 tggcaacccc tctaaccaat cagaagtaag cttgttggaa gtccacagcc taccacttga
     5881 aagcgggctg cacaaaatct gtcaaacgtt ttgaacgttg gatgtgagag cacatacttt
     5941 tattaaggca tctggttggt cagtttataa tatcaacaac ttgggctaca gaaaagaaaa
     6001 gttattacag aaattatgat taacaagtac atgttaaata aagattttaa tatgaatgcc
     6061 accactggag cattcatgcc atttggagct tcttcctgtt tggatcacta gaaggaggag
     6121 gtcactcatt acagttctca tatacagtcg ttggttggtt ggttggttgg tagattgatt
     6181 gattgattga ttgattgatt gattgattga ttgattgatt gattgattga ttgattgatt
     6241 gattgattgg tagtcaaaat aagaaataat ttccacagat tcattacaga aatgattaaa
     6301 tgcatacata aaaaactggg gggggggggg gatacaacaa cacacttaag tcacatttgc
     6361 ctacgtaaag aaaagtaaag aaaatcaata gctatatttt acatctcctt tttttgctgt
     6421 cttttaatta gccttgtttt gctgttatct tgattgaaac tgtaacttct tcacctgctt
     6481 cttttctttg taggttttgc cagcccagaa acaggcatgg ctgtgcaagg ccagagcacc
     6541 atgcacaatg cgctgcatgt cttcatgaac ggctcaatgt cctcagtcca gggctcagcc
     6601 aacgacccca tcttcctcct tcaccatgct ttcattgaca ggtaacaaac acgtcatgac
     6661 attagactgc acagtttttg acaaagttca tacaatctgt tgtttatagc tgctacaatt
     6721 agtgaagttt gtgaatgtac ttggatgagc agcgaaagat caattgagat caattgttag
     6781 agtttggttg ccctgcagag caaagaacaa aaaataatct ggtggcttta ctgcgtgagg
     6841 ttattattgg tggaatagaa acacaaaaca taattgcatt tatttgttta attttttatc
     6901 ttatcttaac tttcatcttg catatttgtt tcttacatca tttctagcat ctttgagcgc
     6961 tggctaagaa ctcatcagcc tccccggtcc atctacccac gtaccaatgc accaattggc
     7021 cacaatgacg gctactacat ggtgccattc cttcctcttt ataggaatgg agactacctc
     7081 ctgtccaaca atgctcttgg atacgagtac gcctacctgt tggacccagg tcattgcaca
     7141 acaccagaaa tgccctctga tctgcaaaag acgtgaatat ctgttcagac acccatatcc
     7201 actctgttcc ac
//
