LOCUS       sequence-199237-        7354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BLInCR 'Localizer' construct that marks the nucleoli and is targeted
            by a PHR-tagged effector upon illumination with blue light.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7354)
  AUTHORS   Rademacher A, Erdel F, Trojanowski J, Schumacher S, Rippe K
  TITLE     Real-time observation of light-controlled transcription in living 
            cells.
  JOURNAL   J Cell Sci. 2017 Dec 15;130(24):4213-4224. doi: 10.1242/jcs.205534. 
            Epub 2017 Nov 9.
  PUBMED    29122982
REFERENCE   2  (bases 1 to 7354)
  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"; doi: 
            "10.1242/jcs.205534"; journalName: "J Cell Sci"; date: "2017-12-15- 
            15"; volume: "130"; issue: "24"; pages: "4213-4224"
COMMENT     Sequence Label: pCIBN-tagBFP-hNCL
FEATURES             Location/Qualifiers
     source          1..7354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        59..362
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        363..566
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     517..537
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             605..1114
                     /codon_start=1
                     /product="N-terminal portion of the CIB1 transcription 
                     factor from Arabidopsis thaliana"
                     /label=CIBN
                     /note="binds to CRY2 (cryptochrome 2) that has absorbed 
                     blue light (Kennedy et al., 2010)"
                     /translation="MNGAIGGDLLLNFPDMSVLERQRAHLKYLNPTFDSPLAGFFADSS
                     MITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKAAKFDTETKDCN
                     EAAKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKE
                     LEKTDYI"
     CDS             1133..1831
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     protein_bind    2441..2452
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     protein_bind    2459..2470
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     CDS             3518..3529
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4140..4261
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4177..4196)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4231..4250
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4268..4723)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4405..4426)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4617..4636
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4750..4854
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4856..5213
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4856..5047
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4879..4899)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5064..5199
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5248..6042
                     /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             5248..6042
                     /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(5302..5321)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5912..5931
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6230..6249)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6274..6321
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6650..7238
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7139..7158
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
       61 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      121 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      181 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      241 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      301 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      361 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      421 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      481 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      541 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctcgc
      601 caccatgaat ggagctatag gaggtgacct tttgctcaat tttcctgaca tgtcggtcct
      661 agagcgccaa agggctcacc tcaagtacct caatcccacc tttgattctc ctctcgccgg
      721 cttctttgcc gattcttcaa tgattaccgg cggcgagatg gacagctatc tttcgactgc
      781 cggtttgaat cttccgatga tgtacggtga gacgacggtg gaaggtgatt caagactctc
      841 aatttcgccg gaaacgacgc ttgggactgg aaatttcaag gcagcgaagt ttgatacaga
      901 gactaaggat tgtaatgagg cggcgaagaa gatgacgatg aacagagatg acctagtaga
      961 agaaggagaa gaagagaagt cgaaaataac agagcaaaac aatgggagca caaaaagcat
     1021 caagaagatg aaacacaaag ccaagaaaga agagaacaat ttctctaatg attcatctaa
     1081 agtgacgaag gaattggaga aaacggatta tattcatgga ccggtcgcca ccatgagcga
     1141 gctgattaag gagaacatgc acatgaagct gtacatggag ggcaccgtgg acaaccatca
     1201 cttcaagtgc acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgagaat
     1261 caaggtggtc gagggcggcc ctctcccctt cgccttcgac atcctggcta ctagcttcct
     1321 ctacggcagc aagaccttca tcaaccacac ccagggcatc cccgacttct tcaagcagtc
     1381 cttccctgag ggcttcacat gggagagagt caccacatac gaagacgggg gcgtgctgac
     1441 cgctacccag gacaccagcc tccaggacgg ctgcctcatc tacaacgtca agatcagagg
     1501 ggtgaacttc acatccaacg gccctgtgat gcagaagaaa acactcggct gggaggcctt
     1561 caccgagacg ctgtaccccg ctgacggcgg cctggaaggc agaaacgaca tggccctgaa
     1621 gctcgtgggc gggagccatc tgatcgcaaa catcaagacc acatatagat ccaagaaacc
     1681 cgctaagaac ctcaagatgc ctggcgtcta ctatgtggac tacagactgg aaagaatcaa
     1741 ggaggccaac aacgagacct acgtcgagca gcacgaggtg gcagtggcca gatactgcga
     1801 cctccctagc aaactggggc acaagcttaa ttccggactc agatctcgag ctcaagcttc
     1861 gaattctgca gtcgacatgg tgaagctcgc gaaggcaggt aaaaatcaag gtgaccccaa
     1921 gaaaatggct cctcctccaa aggaggtaga agaagatagt gaagatgagg aaatgtcaga
     1981 agatgaagaa gatgatagca gtggagaaga ggtcgtcata cctcagaaga aaggcaagaa
     2041 ggctgctgca acctcagcaa agaaggtggt cgtttcccca acaaaaaagg ttgcagttgc
     2101 cacaccagcc aagaaagcag ctgtcactcc aggcaaaaag gcagcagcaa cacctgccaa
     2161 gaagacagtt acaccagcca aagcagttac cacacctggc aagaagggag ccacaccagg
     2221 caaagcattg gtagcaactc ctggtaagaa gggtgctgcc atcccagcca ggggggcaaa
     2281 gaatggcaag aatgccgagg aggaagacag tgatgaagag gaggatgatg acagtgagga
     2341 ggatgaggag gatgacgagg acgaggatga ggatgaagat gaaattgaac cagcagcgat
     2401 gaaagcagca gctgctgccc ctgcctcaga ggatgaggac gatgaggatg acgaagatga
     2461 tgaggatgac gatgacgatg aggaagatga ctctgaagaa gaagctatgg agactacacc
     2521 agccaaagga aagaaagctg caaaagttgt tcctgtgaaa gccaagaacg tggctgagga
     2581 tgaagatgaa gaagaggatg atgaggacga ggatgacgac gacgacgaag atgatgaaga
     2641 tgatgatgat gaagatgatg aggaggagga agaagaggag gaggaagagc ctgtcaaaga
     2701 agcacctgga aaacgaaaga aggaaatggc caaacagaaa gcagctcctg aagccaagaa
     2761 acagaaagtg gaaggcacag aaccgactac ggctttcaat ctctttgttg gaaacctaaa
     2821 ctttaacaaa tctgctcctg aattaaaaac tggtatcagc gatgtttttg ctaaaaatga
     2881 tcttgctgtt gtggatgtca gaattggtat gactaggaaa tttggttatg tggattttga
     2941 atctgctgaa gacctggaga aagcgttgga actcactggt ttgaaagtct ttggcaatga
     3001 aattaaacta gagaaaccaa aaggaaaaga cagtaagaaa gagcgagatg cgagaacact
     3061 tttggctaaa aatctccctt acaaagtcac tcaggatgaa ttgaaagaag tgtttgaaga
     3121 tgctgcggag atcagattag tcagcaagga tgggaaaagt aaagggattg cttatattga
     3181 atttaagaca gaagctgatg cagagaaaac ctttgaagaa aagcagggaa cagagatcga
     3241 tgggcgatct atttccctgt actatactgg agagaaaggt caaaatcaag actatagagg
     3301 tggaaagaat agcacttgga gtggtgaatc aaaaactctg gttttaagca acctctccta
     3361 cagtgcaaca gaagaaactc ttcaggaagt atttgagaaa gcaactttta tcaaagtacc
     3421 ccagaaccaa aatggcaaat ctaaagggta tgcatttata gagtttgctt cattcgaaga
     3481 cgctaaagaa gctttaaatt cctgtaataa aagggaaatt gagggcagag caatcaggct
     3541 ggagttgcaa ggacccaggg gatcacctaa tgccagaagc cagccatcca aaactctgtt
     3601 tgtcaaaggc ctgtctgagg ataccactga agagacatta aaggagtcat ttgacggctc
     3661 cgttcgggct aggatagtta ctgaccggga aactgggtcc tccaaagggt ttggttttgt
     3721 agacttcaac agtgaggagg atgccaaagc tgccaaggag gccatggaag acggtgaaat
     3781 tgatggaaat aaagttacct tggactgggc caaacctaag ggtgaaggtg gcttcggggg
     3841 tcgtggtgga ggcagaggcg gctttggagg acgaggtggt ggtagaggag gccgaggagg
     3901 atttggtggc agaagccggg gaggctttgg agggcgagga ggcttccgag gaggcagagg
     3961 aggaggaggt gaccacaagc cacaaggaaa gaagacgaag tttgaatagg ggatccaccg
     4021 gatctagata actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     4081 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     4141 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4201 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4261 aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     4321 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     4381 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     4441 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     4501 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     4561 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     4621 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     4681 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     4741 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4801 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     4861 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4921 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4981 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5041 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     5101 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     5161 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
     5221 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     5281 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     5341 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     5401 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     5461 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     5521 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     5581 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5641 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5701 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5761 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5821 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5881 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5941 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     6001 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     6061 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     6121 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     6181 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     6241 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     6301 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     6361 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     6421 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     6481 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6541 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6601 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6661 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6721 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6781 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6841 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6901 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6961 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7021 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7081 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7141 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     7201 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     7261 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     7321 attctgtgga taaccgtatt accgccatgc atta
//
