LOCUS       sequence-201623-        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        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             607..1116
                     /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             1135..1833
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     protein_bind    2443..2454
                     /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    2461..2472
                     /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             3520..3531
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4142..4263
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4179..4198)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4233..4252
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4270..4725)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4407..4428)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4619..4638
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4752..4856
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4858..5215
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4858..5049
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4881..4901)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5066..5201
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5250..6044
                     /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             5250..6044
                     /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(5304..5323)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5914..5933
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6232..6251)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6276..6323
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6652..7240
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7141..7160
                     /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 gctagcgctc
      601 gccaccatga atggagctat aggaggtgac cttttgctca attttcctga catgtcggtc
      661 ctagagcgcc aaagggctca cctcaagtac ctcaatccca cctttgattc tcctctcgcc
      721 ggcttctttg ccgattcttc aatgattacc ggcggcgaga tggacagcta tctttcgact
      781 gccggtttga atcttccgat gatgtacggt gagacgacgg tggaaggtga ttcaagactc
      841 tcaatttcgc cggaaacgac gcttgggact ggaaatttca aggcagcgaa gtttgataca
      901 gagactaagg attgtaatga ggcggcgaag aagatgacga tgaacagaga tgacctagta
      961 gaagaaggag aagaagagaa gtcgaaaata acagagcaaa acaatgggag cacaaaaagc
     1021 atcaagaaga tgaaacacaa agccaagaaa gaagagaaca atttctctaa tgattcatct
     1081 aaagtgacga aggaattgga gaaaacggat tatattcatg gaccggtcgc caccatgagc
     1141 gagctgatta aggagaacat gcacatgaag ctgtacatgg agggcaccgt ggacaaccat
     1201 cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga
     1261 atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc tactagcttc
     1321 ctctacggca gcaagacctt catcaaccac acccagggca tccccgactt cttcaagcag
     1381 tccttccctg agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg
     1441 accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga
     1501 ggggtgaact tcacatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc
     1561 ttcaccgaga cgctgtaccc cgctgacggc ggcctggaag gcagaaacga catggccctg
     1621 aagctcgtgg gcgggagcca tctgatcgca aacatcaaga ccacatatag atccaagaaa
     1681 cccgctaaga acctcaagat gcctggcgtc tactatgtgg actacagact ggaaagaatc
     1741 aaggaggcca acaacgagac ctacgtcgag cagcacgagg tggcagtggc cagatactgc
     1801 gacctcccta gcaaactggg gcacaagctt aattccggac tcagatctcg agctcaagct
     1861 tcgaattctg cagtcgacat ggtgaagctc gcgaaggcag gtaaaaatca aggtgacccc
     1921 aagaaaatgg ctcctcctcc aaaggaggta gaagaagata gtgaagatga ggaaatgtca
     1981 gaagatgaag aagatgatag cagtggagaa gaggtcgtca tacctcagaa gaaaggcaag
     2041 aaggctgctg caacctcagc aaagaaggtg gtcgtttccc caacaaaaaa ggttgcagtt
     2101 gccacaccag ccaagaaagc agctgtcact ccaggcaaaa aggcagcagc aacacctgcc
     2161 aagaagacag ttacaccagc caaagcagtt accacacctg gcaagaaggg agccacacca
     2221 ggcaaagcat tggtagcaac tcctggtaag aagggtgctg ccatcccagc caggggggca
     2281 aagaatggca agaatgccga ggaggaagac agtgatgaag aggaggatga tgacagtgag
     2341 gaggatgagg aggatgacga ggacgaggat gaggatgaag atgaaattga accagcagcg
     2401 atgaaagcag cagctgctgc ccctgcctca gaggatgagg acgatgagga tgacgaagat
     2461 gatgaggatg acgatgacga tgaggaagat gactctgaag aagaagctat ggagactaca
     2521 ccagccaaag gaaagaaagc tgcaaaagtt gttcctgtga aagccaagaa cgtggctgag
     2581 gatgaagatg aagaagagga tgatgaggac gaggatgacg acgacgacga agatgatgaa
     2641 gatgatgatg atgaagatga tgaggaggag gaagaagagg aggaggaaga gcctgtcaaa
     2701 gaagcacctg gaaaacgaaa gaaggaaatg gccaaacaga aagcagctcc tgaagccaag
     2761 aaacagaaag tggaaggcac agaaccgact acggctttca atctctttgt tggaaaccta
     2821 aactttaaca aatctgctcc tgaattaaaa actggtatca gcgatgtttt tgctaaaaat
     2881 gatcttgctg ttgtggatgt cagaattggt atgactagga aatttggtta tgtggatttt
     2941 gaatctgctg aagacctgga gaaagcgttg gaactcactg gtttgaaagt ctttggcaat
     3001 gaaattaaac tagagaaacc aaaaggaaaa gacagtaaga aagagcgaga tgcgagaaca
     3061 cttttggcta aaaatctccc ttacaaagtc actcaggatg aattgaaaga agtgtttgaa
     3121 gatgctgcgg agatcagatt agtcagcaag gatgggaaaa gtaaagggat tgcttatatt
     3181 gaatttaaga cagaagctga tgcagagaaa acctttgaag aaaagcaggg aacagagatc
     3241 gatgggcgat ctatttccct gtactatact ggagagaaag gtcaaaatca agactataga
     3301 ggtggaaaga atagcacttg gagtggtgaa tcaaaaactc tggttttaag caacctctcc
     3361 tacagtgcaa cagaagaaac tcttcaggaa gtatttgaga aagcaacttt tatcaaagta
     3421 ccccagaacc aaaatggcaa atctaaaggg tatgcattta tagagtttgc ttcattcgaa
     3481 gacgctaaag aagctttaaa ttcctgtaat aaaagggaaa ttgagggcag agcaatcagg
     3541 ctggagttgc aaggacccag gggatcacct aatgccagaa gccagccatc caaaactctg
     3601 tttgtcaaag gcctgtctga ggataccact gaagagacat taaaggagtc atttgacggc
     3661 tccgttcggg ctaggatagt tactgaccgg gaaactgggt cctccaaagg gtttggtttt
     3721 gtagacttca acagtgagga ggatgccaaa gctgccaagg aggccatgga agacggtgaa
     3781 attgatggaa ataaagttac cttggactgg gccaaaccta agggtgaagg tggcttcggg
     3841 ggtcgtggtg gaggcagagg cggctttgga ggacgaggtg gtggtagagg aggccgagga
     3901 ggatttggtg gcagaagccg gggaggcttt ggagggcgag gaggcttccg aggaggcaga
     3961 ggaggaggag gtgaccacaa gccacaagga aagaagacga agtttgaata ggggatccac
     4021 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     4081 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     4141 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4201 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4261 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     4321 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     4381 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     4441 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     4501 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     4561 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     4621 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     4681 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     4741 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4801 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     4861 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4921 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4981 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5041 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     5101 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     5161 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     5221 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     5281 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     5341 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5401 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     5461 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5521 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5581 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5641 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5701 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5761 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5821 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5881 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5941 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     6001 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     6061 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     6121 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     6181 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     6241 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     6301 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     6361 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     6421 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     6481 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     6541 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6601 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6661 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6721 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6781 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6841 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6901 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6961 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7021 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     7081 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7141 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7201 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7261 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     7321 tgattctgtg gataaccgta ttaccgccat gcat
//
