LOCUS       sequence-199242-        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        65..368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        369..572
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     523..543
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             611..1120
                     /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      1133..1142
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1139..1141,1142..1144,1145..1855)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1184..1205)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1445..1464)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1792..1813
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3957..4078
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3994..4013)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4048..4067
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4085..4540)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4222..4243)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4434..4453
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4567..4671
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4673..5030
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4673..4864
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4696..4716)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4881..5016
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5065..5859
                     /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             5065..5859
                     /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(5119..5138)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5729..5748
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6047..6066)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6091..6138
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6467..7055
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6956..6975
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
       61 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      121 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      181 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      241 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      301 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      361 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      421 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      481 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      541 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc
      601 gctcgccacc atgaatggag ctataggagg tgaccttttg ctcaattttc ctgacatgtc
      661 ggtcctagag cgccaaaggg ctcacctcaa gtacctcaat cccacctttg attctcctct
      721 cgccggcttc tttgccgatt cttcaatgat taccggcggc gagatggaca gctatctttc
      781 gactgccggt ttgaatcttc cgatgatgta cggtgagacg acggtggaag gtgattcaag
      841 actctcaatt tcgccggaaa cgacgcttgg gactggaaat ttcaaggcag cgaagtttga
      901 tacagagact aaggattgta atgaggcggc gaagaagatg acgatgaaca gagatgacct
      961 agtagaagaa ggagaagaag agaagtcgaa aataacagag caaaacaatg ggagcacaaa
     1021 aagcatcaag aagatgaaac acaaagccaa gaaagaagag aacaatttct ctaatgattc
     1081 atctaaagtg acgaaggaat tggagaaaac ggattatatt catggaccgg tcgccaccat
     1141 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1201 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1261 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1321 cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca
     1381 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1441 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1501 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1561 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     1621 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     1681 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1741 cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1801 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg
     1861 actcagatct cgagctcaag cttcgaattc tgcagtcgac atggagcctg cacccgcccg
     1921 atctccgagg ccccagcagg accccgcccg gccccaggag cccaccatgc ctccccccga
     1981 gaccccctct gaaggccgcc agcccagccc cagccccagc cctacagagc gagcccccgc
     2041 ttcggaggag gagttccagt ttctgcgctg ccagcaatgc caggcggaag ccaagtgccc
     2101 gaagctgctg ccttgtctgc acacgctgtg ctcaggatgc ctggaggcgt cgggcatgca
     2161 gtgccccatc tgccaggcgc cctggcccct aggtgcagac acacccgccc tggataacgt
     2221 ctttttcgag agtctgcagc ggcgcctgtc ggtgtaccgg cagattgtgg atgcgcaggc
     2281 tgtgtgcacc cgctgcaaag agtcggccga cttctggtgc tttgagtgcg agcagctcct
     2341 ctgcgccaag tgcttcgagg cacaccagtg gttcctcaag cacgaggccc ggcccctagc
     2401 agagctgcgc aaccagtcgg tgcgtgagtt cctggacggc acccgcaaga ccaacaacat
     2461 cttctgctcc aaccccaacc accgcacccc tacgctgacc agcatctact gccgaggatg
     2521 ttccaagccg ctgtgctgct cgtgcgcgct ccttgacagc agccacagtg agctcaagtg
     2581 cgacatcagc gcagagatcc agcagcgaca ggaggagctg gacgccatga cgcaggcgct
     2641 gcaggagcag gatagtgcct ttggcgcggt tcacgcgcag atgcacgcgg ccgtcggcca
     2701 gctgggccgc gcgcgtgccg agaccgagga gctgatccgc gagcgcgtgc gccaggtggt
     2761 agctcacgtg cgggctcagg agcgcgagct gctggaggct gtggacgcgc ggtaccagcg
     2821 cgactacgag gagatggcca gtcggctggg ccgcctggat gctgtgctgc agcgcatccg
     2881 cacgggcagc gcgctggtgc agaggatgaa gtgctacgcc tcggaccagg aggtgctgga
     2941 catgcacggt ttcctgcgcc aggcgctctg ccgcctgcgc caggaggagc cccagagcct
     3001 gcaagctgcc gtgcgcaccg atggcttcga cgagttcaag gtgcgcctgc aggacctcag
     3061 ctcttgcatc acccagggga aagatgcagc tgtatccaag aaagccagcc cagaggctgc
     3121 cagcactccc agggacccta ttgacgttga cctgcccgag gaggcagaga gagtgaaggc
     3181 ccaggttcag gccctggggc tggctgaagc ccagcctatg gctgtggtac agtcagtgcc
     3241 cggggcacac cccgtgccag tgtacgcctt ctccatcaaa ggcccttcct atggagagga
     3301 tgtctccaat acaacgacag cccagaagag gaagtgcagc cagacccagt gccccaggaa
     3361 ggtcatcaag atggagtctg aggaggggaa ggaggcaagg ttggctcgga gctccccgga
     3421 gcagcccagg cccagcacct ccaaggcagt ctcaccaccc cacctggatg gaccgcctag
     3481 ccccaggagc cccgtcatag gaagtgaggt cttcctgccc aacagcaacc acgtggccag
     3541 tggcgccggg gaggcagagg aacgcgttgt ggtgatcagc agctcggaag actcagatgc
     3601 cgaaaactcg gtctcttcca gccctcagtc tgaggttctg tattggaaag tgcatggagc
     3661 ccatggagac cgccgagcca cagtcctcgc cagcccactc ctcgccagcc cactcctcgc
     3721 cagcccactc ctcgccagtc cagtctctgc tgagagcaca aggagcctcc agcctgccct
     3781 gtggcacata ccacccccca gcttggcctc cccaccagcc cgctgaggga tccaccggat
     3841 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     3901 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     3961 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4021 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     4081 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     4141 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     4201 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     4261 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     4321 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     4381 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     4441 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     4501 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     4561 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4621 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     4681 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4741 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4801 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4861 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4921 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     4981 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     5041 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     5101 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5161 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5221 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     5281 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5341 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5401 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5461 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5521 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5581 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5641 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5701 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5761 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5821 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5881 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5941 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     6001 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     6061 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     6121 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     6181 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     6241 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     6301 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     6361 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     6421 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6481 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6541 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6601 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6661 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6721 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6781 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6841 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6901 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6961 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     7021 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     7081 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     7141 ctgtggataa ccgtattacc gccat
//
