LOCUS       sequence-414204-        6913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of WDR5 with N terminal RBP fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6913)
  AUTHORS   Sekhon H, Ha JH, Presti MF, Procopio SB, Jarvis AR, Mirsky PO, John 
            AM, Loh SN
  TITLE     Adaptable, turn-on maturation (ATOM) fluorescent biosensors for 
            multiplexed detection in cells.
  JOURNAL   Nat Methods. 2023 Nov 9. doi: 10.1038/s41592-023-02065-w.
  PUBMED    37945909
REFERENCE   2  (bases 1 to 6913)
  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 
            Methods. 2023 Nov 9. doi: 10.1038/s41592-023-02065-w."
COMMENT     Sequence Label: pCMV-RBP-WDR5
FEATURES             Location/Qualifiers
     source          1..6913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1524..1547
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     regulatory      2665..2674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2671..3369
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     polyA_signal    3701..3822
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3738..3757)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3792..3811
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3829..4284)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3966..3987)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4178..4197
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4311..4415
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4417..4774
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4417..4608
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4440..4460)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4625..4760
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4809..5603
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4809..5603
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4863..4882)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5473..5492
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5791..5810)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5835..5882
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6211..6799
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6700..6719
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgccac catgaaagag ggcaaaacga
      661 ttggcctggt gatctctacc ctgaacaatc cgttctttgt gaccctgaaa aatggtgcgg
      721 aagaaaaagc gaaagaactg ggttacaaaa ttatcgttga agattcgcaa aatgattcct
      781 ctaaagagct gtctaatgtc gaagatttga ttcaacagaa agttgatgtt ctgctgatca
      841 atccggtgga tagcgatgcg gttgttacgg cgattaaaga agcgaatagc aaaaatatcc
      901 cggttattac catcgatcgc agcgcgaatg gtggtgatgt tgtttcccat atcgccagcg
      961 ataatgttaa gggtggcgaa atggccgcgg aatttatcgc gaaagccctg aaaggcaagg
     1021 ggaatgttgt ggaactggaa ggtatcccgg gggcgtctgc ggcacgtgat cgcggcaaag
     1081 ggtttgatga agccattgct aagtatccgg atattaaaat cgttgcaaag caggcggcgg
     1141 attttgatcg ttccaaaggt ctgtcagtga tggaaaacat cttgcaagcc cagccgaaaa
     1201 ttgatgcagt gtttgcgcaa aatgatgaaa tggctctggg cgctatcaaa gccattgagg
     1261 ccgcgaatcg tcaaggtatt attgttgtgg gctttgatgg gaccgaagat gctctgaaag
     1321 cgattaaaga agggaaaatg gctgcgacca ttgcgcagca gccggccctg atgggctcac
     1381 tgggtgtgga gatggctgat aaatacctga aaggtgaaaa aattccgaac tttattccgg
     1441 cagaactgaa actcatcacg aaagaaaatg tgcagggtgg agcggcaagc gggggtgccg
     1501 cgggtggcag ctctgcggcc gcattagaag tgctgtttca aggtccaggc atggtgccca
     1561 tcggagccgt gcacggcggc catcccggcg tagtgcatcc gccacagcaa ccactgccca
     1621 cggcgcccag cggcccaaac tcgctgcagc cgaactcggt gggccagccg ggggccacca
     1681 cctcctcgaa cagcagcgcc tccaacaaga gctcgctatc cgtcaagccc aactacacgc
     1741 tcaagttcac gctggccggg cacaccaagg cggtgtcggc ggtcaagttc agtccgaatg
     1801 gcgagtggct ggccagctcc tccgctgata aactaatcaa aatctgggga gcatacgatg
     1861 gcaagttcga gaagaccatt tcgggccaca agctgggcat cagcgatgtg gcctggagct
     1921 cagactcgcg actcctcgtg agcggcagtg atgacaagac gctcaaggtc tgggagctga
     1981 gcaccgggaa gagcttgaaa actctgaagg gccacagcaa ctatgtgttc tgctgcaact
     2041 ttaatccgca gtccaatctg atcgtctccg gcagcttcga cgagagcgtt cgcatatggg
     2101 atgtgcgcac cggcaagtgt ctgaagactc tacccgccca ttccgatccc gtttcggcgg
     2161 tacatttcaa tcgcgacgga tcgctgatcg tgagcagcag ctacgacggc ctctgtcgca
     2221 tatgggacac ggccagtgga cagtgcttga aaaccctgat cgacgacgac aatccgcccg
     2281 tcagctttgt aaagttctcg cccaatggca agtacatttt ggccgccacg ctggataata
     2341 cgctcaagtt gtgggactac tcgaagggca agtgcctgaa gacgtatacg ggtcacaaga
     2401 atgagaagta ctgcatattc gccaacttct cggtgacggg aggaaagtgg atcgtgagtg
     2461 gcagcgagga caacatggtc tacatttgga atctgcagag caaggaggtg gtgcaaaagc
     2521 tgcagggaca caccgatacc gttctgtgca ccgcctgcca tcccacggag aacatcattg
     2581 cttccgcggc gctcgagaac gacaagacca tcaagctgtg gaagtcggat acatagggta
     2641 ccgcgggccc gggatccacc ggtcgccacc atggtgagcg agctgattaa ggagaacatg
     2701 cacatgaagc tgtacatgga gggcaccgtg aacaaccacc acttcaagtg cacatccgag
     2761 ggcgaaggca agccctacga gggcacccag accatgagaa tcaaggcggt cgagggcggc
     2821 cctctcccct tcgccttcga catcctggct accagcttca tgtacggcag caaaaccttc
     2881 atcaaccaca cccagggcat ccccgacttc tttaagcagt ccttccccga gggcttcaca
     2941 tgggagagag tcaccacata cgaagacggg ggcgtgctga ccgctaccca ggacaccagc
     3001 ctccaggacg gctgcctcat ctacaacgtc aagatcagag gggtgaactt cccatccaac
     3061 ggccctgtga tgcagaagaa aacactcggc tgggaggcct ccaccgagac cctgtacccc
     3121 gctgacggcg gcctggaagg cagagccgac atggccctga agctcgtggg cgggggccac
     3181 ctgatctgca acttgaagac cacatacaga tccaagaaac ccgctaagaa cctcaagatg
     3241 cccggcgtct actatgtgga cagaagactg gaaagaatca aggaggccga caaagagacc
     3301 tacgtcgagc agcacgaggt ggctgtggcc agatactgcg acctccctag caaactgggg
     3361 cacagatgag cggccgcttg ccatgtgtat gtgggagacg gtcgggtcca tctgagacgg
     3421 tcgggtccag atattcgtat ctgtcgagta gagtgtgggc tcagatgtcg agtagagtgt
     3481 gggctggatc attcatggca atctagagcg gacttcggtc cgctttttac taggacctgc
     3541 aggcatgcaa gcttgacgtc ggttaccgat atccatatgg cggccgcgac tctagatcat
     3601 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3661 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3721 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3781 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     3841 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3901 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3961 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     4021 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     4081 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     4141 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     4201 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     4261 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4321 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4381 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     4441 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4501 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4561 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4621 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4681 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4741 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     4801 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4861 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4921 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4981 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     5041 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     5101 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     5161 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     5221 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     5281 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     5341 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5401 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5461 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5521 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5581 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     5641 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5701 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5761 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     5821 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     5881 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     5941 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     6001 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     6061 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6121 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6181 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6241 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6301 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6361 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6421 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6481 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6541 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6601 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6661 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6721 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6781 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6841 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6901 taccgccatg cat
//
