LOCUS       sequence-199241-        7147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BLInCR 'Localizer' construct that marks the PML nuclear bodies 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 7147)
  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 7147)
  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-hPMLIII
FEATURES             Location/Qualifiers
     source          1..7147
                     /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"
     polyA_signal    3933..4054
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3970..3989)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4024..4043
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4061..4516)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4198..4219)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4410..4429
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4543..4647
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4649..5006
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4649..4840
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4672..4692)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4857..4992
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5041..5835
                     /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             5041..5835
                     /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(5095..5114)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5705..5724
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6023..6042)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6067..6114
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6443..7031
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6932..6951
                     /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 agcctgcacc cgcccgatct ccgaggcccc agcaggaccc
     1921 cgcccggccc caggagccca ccatgcctcc ccccgagacc ccctctgaag gccgccagcc
     1981 cagccccagc cccagcccta cagagcgagc ccccgcttcg gaggaggagt tccagtttct
     2041 gcgctgccag caatgccagg cggaagccaa gtgcccgaag ctgctgcctt gtctgcacac
     2101 gctgtgctca ggatgcctgg aggcgtcggg catgcagtgc cccatctgcc aggcgccctg
     2161 gcccctaggt gcagacacac ccgccctgga taacgtcttt ttcgagagtc tgcagcggcg
     2221 cctgtcggtg taccggcaga ttgtggatgc gcaggctgtg tgcacccgct gcaaagagtc
     2281 ggccgacttc tggtgctttg agtgcgagca gctcctctgc gccaagtgct tcgaggcaca
     2341 ccagtggttc ctcaagcacg aggcccggcc cctagcagag ctgcgcaacc agtcggtgcg
     2401 tgagttcctg gacggcaccc gcaagaccaa caacatcttc tgctccaacc ccaaccaccg
     2461 cacccctacg ctgaccagca tctactgccg aggatgttcc aagccgctgt gctgctcgtg
     2521 cgcgctcctt gacagcagcc acagtgagct caagtgcgac atcagcgcag agatccagca
     2581 gcgacaggag gagctggacg ccatgacgca ggcgctgcag gagcaggata gtgcctttgg
     2641 cgcggttcac gcgcagatgc acgcggccgt cggccagctg ggccgcgcgc gtgccgagac
     2701 cgaggagctg atccgcgagc gcgtgcgcca ggtggtagct cacgtgcggg ctcaggagcg
     2761 cgagctgctg gaggctgtgg acgcgcggta ccagcgcgac tacgaggaga tggccagtcg
     2821 gctgggccgc ctggatgctg tgctgcagcg catccgcacg ggcagcgcgc tggtgcagag
     2881 gatgaagtgc tacgcctcgg accaggaggt gctggacatg cacggtttcc tgcgccaggc
     2941 gctctgccgc ctgcgccagg aggagcccca gagcctgcaa gctgccgtgc gcaccgatgg
     3001 cttcgacgag ttcaaggtgc gcctgcagga cctcagctct tgcatcaccc aggggaaaga
     3061 tgcagctgta tccaagaaag ccagcccaga ggctgccagc actcccaggg accctattga
     3121 cgttgacctg cccgaggagg cagagagagt gaaggcccag gttcaggccc tggggctggc
     3181 tgaagcccag cctatggctg tggtacagtc agtgcccggg gcacaccccg tgccagtgta
     3241 cgccttctcc atcaaaggcc cttcctatgg agaggatgtc tccaatacaa cgacagccca
     3301 gaagaggaag tgcagccaga cccagtgccc caggaaggtc atcaagatgg agtctgagga
     3361 ggggaaggag gcaaggttgg ctcggagctc cccggagcag cccaggccca gcacctccaa
     3421 ggcagtctca ccaccccacc tggatggacc gcctagcccc aggagccccg tcataggaag
     3481 tgaggtcttc ctgcccaaca gcaaccacgt ggccagtggc gccggggagg cagaggaacg
     3541 cgttgtggtg atcagcagct cggaagactc agatgccgaa aactcggtct cttccagccc
     3601 tcagtctgag gttctgtatt ggaaagtgca tggagcccat ggagaccgcc gagccacagt
     3661 cctcgccagc ccactcctcg ccagcccact cctcgccagc ccactcctcg ccagtccagt
     3721 ctctgctgag agcacaagga gcctccagcc tgccctgtgg cacataccac cccccagctt
     3781 ggcctcccca ccagcccgct gagggatcca ccggatctag ataactgatc ataatcagcc
     3841 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     3901 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     3961 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4021 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     4081 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     4141 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     4201 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     4261 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     4321 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     4381 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     4441 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     4501 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4561 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4621 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     4681 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4741 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4801 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4861 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4921 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     4981 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     5041 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5101 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5161 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5221 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5281 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5341 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5401 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5461 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5521 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     5581 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5641 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5701 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5761 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5821 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5881 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5941 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     6001 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     6061 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     6121 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     6181 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     6241 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     6301 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6361 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6421 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6481 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6541 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6601 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6661 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6721 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6781 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6841 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6901 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6961 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7021 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7081 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     7141 tgcatta
//
