LOCUS       sequence-196683-        7165 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 7165)
  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 7165)
  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-EYFP-hPMLIII
FEATURES             Location/Qualifiers
     source          1..7165
                     /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"
     regulatory      1129..1138
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1135..1137,1138..1140,1141..1851)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1180..1201)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1441..1460)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1788..1809
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3953..4074
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3990..4009)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4044..4063
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4081..4536)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4218..4239)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4430..4449
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4563..4667
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4669..5026
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4669..4860
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4692..4712)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4877..5012
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5061..5855
                     /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             5061..5855
                     /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(5115..5134)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5725..5744
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6043..6062)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6087..6134
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6463..7051
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6952..6971
                     /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 caccatggtg
     1141 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1201 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     1261 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1321 accaccttcg gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
     1381 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1441 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1501 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1561 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     1621 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     1681 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1741 agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1801 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtccggactc
     1861 agatctcgag ctcaagcttc gaattctgca gtcgacatgg agcctgcacc cgcccgatct
     1921 ccgaggcccc agcaggaccc cgcccggccc caggagccca ccatgcctcc ccccgagacc
     1981 ccctctgaag gccgccagcc cagccccagc cccagcccta cagagcgagc ccccgcttcg
     2041 gaggaggagt tccagtttct gcgctgccag caatgccagg cggaagccaa gtgcccgaag
     2101 ctgctgcctt gtctgcacac gctgtgctca ggatgcctgg aggcgtcggg catgcagtgc
     2161 cccatctgcc aggcgccctg gcccctaggt gcagacacac ccgccctgga taacgtcttt
     2221 ttcgagagtc tgcagcggcg cctgtcggtg taccggcaga ttgtggatgc gcaggctgtg
     2281 tgcacccgct gcaaagagtc ggccgacttc tggtgctttg agtgcgagca gctcctctgc
     2341 gccaagtgct tcgaggcaca ccagtggttc ctcaagcacg aggcccggcc cctagcagag
     2401 ctgcgcaacc agtcggtgcg tgagttcctg gacggcaccc gcaagaccaa caacatcttc
     2461 tgctccaacc ccaaccaccg cacccctacg ctgaccagca tctactgccg aggatgttcc
     2521 aagccgctgt gctgctcgtg cgcgctcctt gacagcagcc acagtgagct caagtgcgac
     2581 atcagcgcag agatccagca gcgacaggag gagctggacg ccatgacgca ggcgctgcag
     2641 gagcaggata gtgcctttgg cgcggttcac gcgcagatgc acgcggccgt cggccagctg
     2701 ggccgcgcgc gtgccgagac cgaggagctg atccgcgagc gcgtgcgcca ggtggtagct
     2761 cacgtgcggg ctcaggagcg cgagctgctg gaggctgtgg acgcgcggta ccagcgcgac
     2821 tacgaggaga tggccagtcg gctgggccgc ctggatgctg tgctgcagcg catccgcacg
     2881 ggcagcgcgc tggtgcagag gatgaagtgc tacgcctcgg accaggaggt gctggacatg
     2941 cacggtttcc tgcgccaggc gctctgccgc ctgcgccagg aggagcccca gagcctgcaa
     3001 gctgccgtgc gcaccgatgg cttcgacgag ttcaaggtgc gcctgcagga cctcagctct
     3061 tgcatcaccc aggggaaaga tgcagctgta tccaagaaag ccagcccaga ggctgccagc
     3121 actcccaggg accctattga cgttgacctg cccgaggagg cagagagagt gaaggcccag
     3181 gttcaggccc tggggctggc tgaagcccag cctatggctg tggtacagtc agtgcccggg
     3241 gcacaccccg tgccagtgta cgccttctcc atcaaaggcc cttcctatgg agaggatgtc
     3301 tccaatacaa cgacagccca gaagaggaag tgcagccaga cccagtgccc caggaaggtc
     3361 atcaagatgg agtctgagga ggggaaggag gcaaggttgg ctcggagctc cccggagcag
     3421 cccaggccca gcacctccaa ggcagtctca ccaccccacc tggatggacc gcctagcccc
     3481 aggagccccg tcataggaag tgaggtcttc ctgcccaaca gcaaccacgt ggccagtggc
     3541 gccggggagg cagaggaacg cgttgtggtg atcagcagct cggaagactc agatgccgaa
     3601 aactcggtct cttccagccc tcagtctgag gttctgtatt ggaaagtgca tggagcccat
     3661 ggagaccgcc gagccacagt cctcgccagc ccactcctcg ccagcccact cctcgccagc
     3721 ccactcctcg ccagtccagt ctctgctgag agcacaagga gcctccagcc tgccctgtgg
     3781 cacataccac cccccagctt ggcctcccca ccagcccgct gagggatcca ccggatctag
     3841 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3901 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3961 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4021 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     4081 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     4141 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     4201 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4261 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     4321 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     4381 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     4441 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     4501 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     4561 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4621 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     4681 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4741 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4801 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4861 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4921 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4981 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     5041 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     5101 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5161 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5221 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     5281 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5341 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5401 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5461 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5521 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5581 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5641 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5701 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5761 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5821 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5881 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5941 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     6001 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     6061 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     6121 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     6181 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     6241 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     6301 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6361 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6421 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6481 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6541 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6601 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6661 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6721 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6781 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6841 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6901 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6961 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7021 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7081 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     7141 ggataaccgt attaccgcca tgcat
//
