LOCUS       sequence-477298-        7320 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destabilized red fluorescent protein (d2mRFP1) under transcriptional
            control of NF-kB activation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7320)
  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 7320)
  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:d2mRFP1
FEATURES             Location/Qualifiers
     source          1..7320
                     /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             997..1671
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             997..1671
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1127..1145)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1395..1414
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1589..1612
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1810..1944
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1834..1853)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1888..1907
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(2383..2570,2571..2582)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2616..2632)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2616..2632)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2669..2689)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2669..2687)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2708..2724)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2708..2724)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2721..2743)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2732..2748
                     /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(2756..2762,2763..2780,2781..2786))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2801..2822
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2939..2956)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3110..3698)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3190..3209)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3869..4660,4661..4729))
                     /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     4492..4511
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4730..4834)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4860..5315)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4997..5018)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5209..5228
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5442..5464
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5456..5473
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5457..5473
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5483..5502
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5483..5501
                     /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 ttcgagatgg cctcctccga ggacgtcatc
     1021 aaggagttca tgcgcttcaa ggtgcgcatg gagggctccg tgaacggcca cgagttcgag
     1081 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
     1141 accaagggcg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt ccagtacggc
     1201 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
     1261 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     1321 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
     1381 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag
     1441 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agatgaggct gaagctgaag
     1501 gacggcggcc actacgacgc cgaggtcaag accacctaca tggccaagaa gcccgtgcag
     1561 ctgcccggcg cctacaagac cgacatcaag ctggacatca cctcccacaa cgaggactac
     1621 accatcgtgg aacagtacga gcgcgccgag ggccgccact ccaccggcgc caagcttagc
     1681 catggcttcc cgccggaggt ggaggagcag gatgatggca cgctgcccat gtcttgtgcc
     1741 caggagagcg ggatggaccg tcaccctgca gcctgtgctt ctgctaggat caatgtgtag
     1801 atcgatgatg atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     1861 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     1921 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     1981 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
     2041 gattatgatc ctctagatca gatctgcgaa gatacggcca cgggtgctct tgatcctgtg
     2101 gctgattttg gactgtgctg ctcgcagctg ctgatgaatc acatacttcc tccattttct
     2161 tccactgatt gactgttata atttccctaa tttccaggtc aaggtgctgt gcattgtggt
     2221 aatagatgtg acatgacgtc acttccaaag gaccaatgaa catgtctgac caatttcata
     2281 taatgtgaaa acgattttca taggcagaat aaataacatt taaattaaac tgggcatcag
     2341 cgcaattcaa ttggtttggt aatagcaagg gaaaatagaa tgaagtgatc tccaaaaaat
     2401 aagtactttt tgactgtaaa taaaattgta aggagtaaaa agtacttttt tttctaaaaa
     2461 aatgtaatta agtaaaagta aaagtattga tttttaattg tactcaagta aagtaaaaat
     2521 ccccaaaaat aatacttaag tacagtaatc aagtaaaatt actcaagtac tttacacctc
     2581 tggttcttga ccccctacct tcagcaagcc cagcagatcc actagttcta gagcggccgc
     2641 caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     2701 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     2761 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     2821 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     2881 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     2941 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3001 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3061 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3121 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3181 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3241 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3301 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3361 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3421 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3481 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3541 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3601 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3661 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3721 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3781 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3841 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3901 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3961 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4021 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4081 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4141 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4201 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4261 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4321 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4381 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4441 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4501 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4561 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4621 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4681 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4741 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4801 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     4861 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     4921 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     4981 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     5041 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     5101 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     5161 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     5221 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     5281 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
     5341 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     5401 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     5461 aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg gtaccaaata
     5521 gtaggaatta cccacctgta caagtgctga aaacttggat gaataagccc gtttgccatt
     5581 ttctactgct attttaaatc ttttctgtat ttgtctgcat ttgtctttac ccttgaataa
     5641 atgttttagt tgtttttttt caattatgac tgtgtttaac agacattatt acaataatat
     5701 gtaatagtag tacacactat tattatgtaa tagtacttgt tgactgtatt tgagaactgg
     5761 accgagtgag tgttacgtca cccattcaaa atgacttact tctggctcca acaaaatgaa
     5821 gttaattcag ttgccatttt tcactgtatg gacatcgccg tgttggagct agacgttgcc
     5881 aataagcaag aaagagccga gatgcgtcga gtctgagtca ccgttcctat ggcaacccct
     5941 ctaaccaatc agaagtaagc ttgttggaag tccacagcct accacttgaa agcgggctgc
     6001 acaaaatctg tcaaacgttt tgaacgttgg atgtgagagc acatactttt attaaggcat
     6061 ctggttggtc agtttataat atcaacaact tgggctacag aaaagaaaag ttattacaga
     6121 aattatgatt aacaagtaca tgttaaataa agattttaat atgaatgcca ccactggagc
     6181 attcatgcca tttggagctt cttcctgttt ggatcactag aaggaggagg tcactcatta
     6241 cagttctcat atacagtcgt tggttggttg gttggttggt agattgattg attgattgat
     6301 tgattgattg attgattgat tgattgattg attgattgat tgattgattg attgattggt
     6361 agtcaaaata agaaataatt tccacagatt cattacagaa atgattaaat gcatacataa
     6421 aaaactgggg ggggggggga tacaacaaca cacttaagtc acatttgcct acgtaaagaa
     6481 aagtaaagaa aatcaatagc tatattttac atctcctttt tttgctgtct tttaattagc
     6541 cttgttttgc tgttatcttg attgaaactg taacttcttc acctgcttct tttctttgta
     6601 ggttttgcca gcccagaaac aggcatggct gtgcaaggcc agagcaccat gcacaatgcg
     6661 ctgcatgtct tcatgaacgg ctcaatgtcc tcagtccagg gctcagccaa cgaccccatc
     6721 ttcctccttc accatgcttt cattgacagg taacaaacac gtcatgacat tagactgcac
     6781 agtttttgac aaagttcata caatctgttg tttatagctg ctacaattag tgaagtttgt
     6841 gaatgtactt ggatgagcag cgaaagatca attgagatca attgttagag tttggttgcc
     6901 ctgcagagca aagaacaaaa aataatctgg tggctttact gcgtgaggtt attattggtg
     6961 gaatagaaac acaaaacata attgcattta tttgtttaat tttttatctt atcttaactt
     7021 tcatcttgca tatttgtttc ttacatcatt tctagcatct ttgagcgctg gctaagaact
     7081 catcagcctc cccggtccat ctacccacgt accaatgcac caattggcca caatgacggc
     7141 tactacatgg tgccattcct tcctctttat aggaatggag actacctcct gtccaacaat
     7201 gctcttggat acgagtacgc ctacctgttg gacccaggtc attgcacaac accagaaatg
     7261 ccctctgatc tgcaaaagac gtgaatatct gttcagacac ccatatccac tctgttccac
//
