LOCUS       sequence-196680-        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        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"
     polyA_signal    3935..4056
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3972..3991)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4026..4045
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4063..4518)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4200..4221)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4412..4431
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4545..4649
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4651..5008
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4651..4842
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4674..4694)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4859..4994
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5043..5837
                     /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             5043..5837
                     /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(5097..5116)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5707..5726
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6025..6044)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6069..6116
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6445..7033
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6934..6953
                     /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 ggagcctgca cccgcccgat ctccgaggcc ccagcaggac
     1921 cccgcccggc cccaggagcc caccatgcct ccccccgaga ccccctctga aggccgccag
     1981 cccagcccca gccccagccc tacagagcga gcccccgctt cggaggagga gttccagttt
     2041 ctgcgctgcc agcaatgcca ggcggaagcc aagtgcccga agctgctgcc ttgtctgcac
     2101 acgctgtgct caggatgcct ggaggcgtcg ggcatgcagt gccccatctg ccaggcgccc
     2161 tggcccctag gtgcagacac acccgccctg gataacgtct ttttcgagag tctgcagcgg
     2221 cgcctgtcgg tgtaccggca gattgtggat gcgcaggctg tgtgcacccg ctgcaaagag
     2281 tcggccgact tctggtgctt tgagtgcgag cagctcctct gcgccaagtg cttcgaggca
     2341 caccagtggt tcctcaagca cgaggcccgg cccctagcag agctgcgcaa ccagtcggtg
     2401 cgtgagttcc tggacggcac ccgcaagacc aacaacatct tctgctccaa ccccaaccac
     2461 cgcaccccta cgctgaccag catctactgc cgaggatgtt ccaagccgct gtgctgctcg
     2521 tgcgcgctcc ttgacagcag ccacagtgag ctcaagtgcg acatcagcgc agagatccag
     2581 cagcgacagg aggagctgga cgccatgacg caggcgctgc aggagcagga tagtgccttt
     2641 ggcgcggttc acgcgcagat gcacgcggcc gtcggccagc tgggccgcgc gcgtgccgag
     2701 accgaggagc tgatccgcga gcgcgtgcgc caggtggtag ctcacgtgcg ggctcaggag
     2761 cgcgagctgc tggaggctgt ggacgcgcgg taccagcgcg actacgagga gatggccagt
     2821 cggctgggcc gcctggatgc tgtgctgcag cgcatccgca cgggcagcgc gctggtgcag
     2881 aggatgaagt gctacgcctc ggaccaggag gtgctggaca tgcacggttt cctgcgccag
     2941 gcgctctgcc gcctgcgcca ggaggagccc cagagcctgc aagctgccgt gcgcaccgat
     3001 ggcttcgacg agttcaaggt gcgcctgcag gacctcagct cttgcatcac ccaggggaaa
     3061 gatgcagctg tatccaagaa agccagccca gaggctgcca gcactcccag ggaccctatt
     3121 gacgttgacc tgcccgagga ggcagagaga gtgaaggccc aggttcaggc cctggggctg
     3181 gctgaagccc agcctatggc tgtggtacag tcagtgcccg gggcacaccc cgtgccagtg
     3241 tacgccttct ccatcaaagg cccttcctat ggagaggatg tctccaatac aacgacagcc
     3301 cagaagagga agtgcagcca gacccagtgc cccaggaagg tcatcaagat ggagtctgag
     3361 gaggggaagg aggcaaggtt ggctcggagc tccccggagc agcccaggcc cagcacctcc
     3421 aaggcagtct caccacccca cctggatgga ccgcctagcc ccaggagccc cgtcatagga
     3481 agtgaggtct tcctgcccaa cagcaaccac gtggccagtg gcgccgggga ggcagaggaa
     3541 cgcgttgtgg tgatcagcag ctcggaagac tcagatgccg aaaactcggt ctcttccagc
     3601 cctcagtctg aggttctgta ttggaaagtg catggagccc atggagaccg ccgagccaca
     3661 gtcctcgcca gcccactcct cgccagccca ctcctcgcca gcccactcct cgccagtcca
     3721 gtctctgctg agagcacaag gagcctccag cctgccctgt ggcacatacc accccccagc
     3781 ttggcctccc caccagcccg ctgagggatc caccggatct agataactga tcataatcag
     3841 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     3901 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     3961 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4021 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     4081 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     4141 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     4201 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     4261 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     4321 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     4381 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     4441 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     4501 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4561 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4621 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     4681 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4741 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4801 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4861 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4921 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     4981 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     5041 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5101 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5161 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5221 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5281 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5341 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5401 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5461 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5521 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     5581 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5641 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5701 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5761 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5821 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5881 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5941 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     6001 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     6061 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     6121 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     6181 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     6241 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     6301 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6361 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6421 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6481 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6541 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     6601 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6661 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6721 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6781 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6841 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6901 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6961 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     7021 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7081 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     7141 catgcat
//
