LOCUS       sequence-199240-        7372 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 7372)
  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 7372)
  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-hNCL
FEATURES             Location/Qualifiers
     source          1..7372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        40..343
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        344..547
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     498..518
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             586..1095
                     /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      1108..1117
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1114..1116,1117..1119,1120..1830)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1159..1180)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1420..1439)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1767..1788
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    2440..2451
                     /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    2458..2469
                     /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             3517..3528
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4139..4260
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4176..4195)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4230..4249
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4267..4722)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4404..4425)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4616..4635
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4749..4853
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4855..5212
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4855..5046
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4878..4898)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5063..5198
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5247..6041
                     /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             5247..6041
                     /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(5301..5320)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5911..5930
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6229..6248)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6273..6320
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6649..7237
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7138..7157
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
       61 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      121 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      181 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      241 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      301 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      361 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      421 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      481 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      541 cagagctggt ttagtgaacc gtcagatccg ctagcgctcg ccaccatgaa tggagctata
      601 ggaggtgacc ttttgctcaa ttttcctgac atgtcggtcc tagagcgcca aagggctcac
      661 ctcaagtacc tcaatcccac ctttgattct cctctcgccg gcttctttgc cgattcttca
      721 atgattaccg gcggcgagat ggacagctat ctttcgactg ccggtttgaa tcttccgatg
      781 atgtacggtg agacgacggt ggaaggtgat tcaagactct caatttcgcc ggaaacgacg
      841 cttgggactg gaaatttcaa ggcagcgaag tttgatacag agactaagga ttgtaatgag
      901 gcggcgaaga agatgacgat gaacagagat gacctagtag aagaaggaga agaagagaag
      961 tcgaaaataa cagagcaaaa caatgggagc acaaaaagca tcaagaagat gaaacacaaa
     1021 gccaagaaag aagagaacaa tttctctaat gattcatcta aagtgacgaa ggaattggag
     1081 aaaacggatt atattcatgg accggtcgcc accatggtga gcaagggcga ggagctgttc
     1141 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1201 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1261 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg
     1321 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1381 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1441 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1501 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1561 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1621 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1681 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc
     1741 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1801 atcactctcg gcatggacga gctgtacaag tccggactca gatctcgagc tcaagcttcg
     1861 aattctgcag tcgacatggt gaagctcgcg aaggcaggta aaaatcaagg tgaccccaag
     1921 aaaatggctc ctcctccaaa ggaggtagaa gaagatagtg aagatgagga aatgtcagaa
     1981 gatgaagaag atgatagcag tggagaagag gtcgtcatac ctcagaagaa aggcaagaag
     2041 gctgctgcaa cctcagcaaa gaaggtggtc gtttccccaa caaaaaaggt tgcagttgcc
     2101 acaccagcca agaaagcagc tgtcactcca ggcaaaaagg cagcagcaac acctgccaag
     2161 aagacagtta caccagccaa agcagttacc acacctggca agaagggagc cacaccaggc
     2221 aaagcattgg tagcaactcc tggtaagaag ggtgctgcca tcccagccag gggggcaaag
     2281 aatggcaaga atgccgagga ggaagacagt gatgaagagg aggatgatga cagtgaggag
     2341 gatgaggagg atgacgagga cgaggatgag gatgaagatg aaattgaacc agcagcgatg
     2401 aaagcagcag ctgctgcccc tgcctcagag gatgaggacg atgaggatga cgaagatgat
     2461 gaggatgacg atgacgatga ggaagatgac tctgaagaag aagctatgga gactacacca
     2521 gccaaaggaa agaaagctgc aaaagttgtt cctgtgaaag ccaagaacgt ggctgaggat
     2581 gaagatgaag aagaggatga tgaggacgag gatgacgacg acgacgaaga tgatgaagat
     2641 gatgatgatg aagatgatga ggaggaggaa gaagaggagg aggaagagcc tgtcaaagaa
     2701 gcacctggaa aacgaaagaa ggaaatggcc aaacagaaag cagctcctga agccaagaaa
     2761 cagaaagtgg aaggcacaga accgactacg gctttcaatc tctttgttgg aaacctaaac
     2821 tttaacaaat ctgctcctga attaaaaact ggtatcagcg atgtttttgc taaaaatgat
     2881 cttgctgttg tggatgtcag aattggtatg actaggaaat ttggttatgt ggattttgaa
     2941 tctgctgaag acctggagaa agcgttggaa ctcactggtt tgaaagtctt tggcaatgaa
     3001 attaaactag agaaaccaaa aggaaaagac agtaagaaag agcgagatgc gagaacactt
     3061 ttggctaaaa atctccctta caaagtcact caggatgaat tgaaagaagt gtttgaagat
     3121 gctgcggaga tcagattagt cagcaaggat gggaaaagta aagggattgc ttatattgaa
     3181 tttaagacag aagctgatgc agagaaaacc tttgaagaaa agcagggaac agagatcgat
     3241 gggcgatcta tttccctgta ctatactgga gagaaaggtc aaaatcaaga ctatagaggt
     3301 ggaaagaata gcacttggag tggtgaatca aaaactctgg ttttaagcaa cctctcctac
     3361 agtgcaacag aagaaactct tcaggaagta tttgagaaag caacttttat caaagtaccc
     3421 cagaaccaaa atggcaaatc taaagggtat gcatttatag agtttgcttc attcgaagac
     3481 gctaaagaag ctttaaattc ctgtaataaa agggaaattg agggcagagc aatcaggctg
     3541 gagttgcaag gacccagggg atcacctaat gccagaagcc agccatccaa aactctgttt
     3601 gtcaaaggcc tgtctgagga taccactgaa gagacattaa aggagtcatt tgacggctcc
     3661 gttcgggcta ggatagttac tgaccgggaa actgggtcct ccaaagggtt tggttttgta
     3721 gacttcaaca gtgaggagga tgccaaagct gccaaggagg ccatggaaga cggtgaaatt
     3781 gatggaaata aagttacctt ggactgggcc aaacctaagg gtgaaggtgg cttcgggggt
     3841 cgtggtggag gcagaggcgg ctttggagga cgaggtggtg gtagaggagg ccgaggagga
     3901 tttggtggca gaagccgggg aggctttgga gggcgaggag gcttccgagg aggcagagga
     3961 ggaggaggtg accacaagcc acaaggaaag aagacgaagt ttgaataggg gatccaccgg
     4021 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     4081 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     4141 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4201 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4261 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     4321 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     4381 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     4441 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     4501 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     4561 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     4621 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4681 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     4741 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4801 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     4861 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4921 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4981 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     5041 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     5101 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     5161 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     5221 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     5281 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     5341 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     5401 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     5461 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     5521 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     5581 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     5641 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5701 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5761 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5821 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5881 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5941 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     6001 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     6061 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     6121 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     6181 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     6241 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     6301 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     6361 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     6421 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     6481 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6541 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6601 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6661 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6721 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6781 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6841 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6901 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6961 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7021 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7081 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7141 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7201 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7261 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     7321 ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca at
//
