LOCUS       sequence-196662-        6480 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BLInCR 'Localizer' construct that marks the telomeres 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 6480)
  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 6480)
  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-hTRF1
FEATURES             Location/Qualifiers
     source          1..6480
                     /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    3268..3389
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3305..3324)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3359..3378
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3396..3851)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3533..3554)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3745..3764
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3878..3982
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3984..4341
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3984..4175
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4007..4027)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4192..4327
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4376..5170
                     /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             4376..5170
                     /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(4430..4449)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5040..5059
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5358..5377)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5402..5449
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5778..6366
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6267..6286
                     /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 ggcggaggat gtttcctcag cggccccgag cccgcggggc
     1921 tgtgcggatg gtagggatgc cgaccctact gaggagcaga tggcagaaac agagagaaac
     1981 gacgaggagc agttcgaatg ccaggaactg ctcgagtgcc aggtgcaggt gggggccccc
     2041 gaggaggagg aggaggagga ggaggacgcg ggcctggtgg ccgaggccga ggccgtggct
     2101 gccggctgga tgctcgattt cctctgcctc tctctttgcc gagctttccg cgacggccgc
     2161 tccgaggact tccgcaggac ccgcaacagc gcagaggcta ttattcatgg actatccagt
     2221 ctaacagctt gccagttgag aacgatatac atatgtcagt ttttgacaag aattgcagca
     2281 ggaaaaaccc ttgatgcaca gtttgaaaat gatgaacgaa ttacaccctt ggaatcagcc
     2341 ctgatgattt ggggttcaat tgaaaaggaa catgacaaac ttcatgaaga aatacagaat
     2401 ttaattaaaa ttcaggctat agctgtttgt atggaaaatg gcaactttaa agaagcagaa
     2461 gaagtctttg aaagaatatt tggtgatcca aattctcata tgcctttcaa aagcaaattg
     2521 cttatgataa tctctcagaa agatacattt cattcctttt ttcaacactt cagctacaac
     2581 cacatgatgg agaaaattaa gagttatgtg aattatgtgc taagtgaaaa atcatcaacc
     2641 tttctaatga aggcagcggc aaaagtagta gaaagcaaaa ggacaagaac aataacttct
     2701 caagataaac ctagtggtaa tgatgttgaa atggaaactg aagctaattt ggatacaaga
     2761 aaaaggtctc acaagaatct tttcttatct aagttgcaac atggaaccca gcaacaagac
     2821 cttaataaga aagaaagaag agtaggaact cctcaaagta caaaaaagaa aaaagaaagc
     2881 agaagagcca ctgaaagcag aatacctgtt tcaaagagtc agccggtaac tcctgaaaaa
     2941 catcgagcta gaaaaagaca ggcatggctt tgggaagaag acaagaattt gagatctggc
     3001 gtgaggaaat atggagaggg aaactggtct aaaatactgt tgcattataa attcaacaac
     3061 cggacaagtg tcatgttaaa agacagatgg aggaccatga agaaactaaa actgatttcc
     3121 tcagacagcg aagactgagg atccaccgga tctagataac tgatcataat cagccatacc
     3181 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     3241 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     3301 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3361 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     3421 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     3481 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     3541 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     3601 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     3661 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     3721 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     3781 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     3841 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3901 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3961 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     4021 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4081 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4141 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4201 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4261 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4321 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     4381 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4441 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4501 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     4561 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4621 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4681 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4741 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4801 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4861 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     4921 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4981 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     5041 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     5101 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     5161 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     5221 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5281 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5341 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     5401 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     5461 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     5521 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     5581 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     5641 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5701 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5761 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5821 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5881 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     5941 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6001 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6061 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6121 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6181 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6241 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6301 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6361 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6421 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
