LOCUS       sequence-196692-        6443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BLInCR 'Localizer' construct that marks telomeres and is targeted by
            a PHR-tagged effector upon illumination with blue light; can be 
            visualized without triggering PHR recruitment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6443)
  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 6443)
  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-hTRF1-tagRFP-T
FEATURES             Location/Qualifiers
     source          1..6443
                     /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             603..1112
                     /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             2388..3101
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     polyA_signal    3231..3352
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3268..3287)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3322..3341
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3359..3814)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3496..3517)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3708..3727
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3841..3945
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3947..4304
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3947..4138
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3970..3990)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4155..4290
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4339..5133
                     /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             4339..5133
                     /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(4393..4412)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5003..5022
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5321..5340)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5365..5412
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5741..6329
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6230..6249
                     /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 gctagcgcca
      601 ccatgaatgg agctatagga ggtgaccttt tgctcaattt tcctgacatg tcggtcctag
      661 agcgccaaag ggctcacctc aagtacctca atcccacctt tgattctcct ctcgccggct
      721 tctttgccga ttcttcaatg attaccggcg gcgagatgga cagctatctt tcgactgccg
      781 gtttgaatct tccgatgatg tacggtgaga cgacggtgga aggtgattca agactctcaa
      841 tttcgccgga aacgacgctt gggactggaa atttcaaggc agcgaagttt gatacagaga
      901 ctaaggattg taatgaggcg gcgaagaaga tgacgatgaa cagagatgac ctagtagaag
      961 aaggagaaga agagaagtcg aaaataacag agcaaaacaa tgggagcaca aaaagcatca
     1021 agaagatgaa acacaaagcc aagaaagaag agaacaattt ctctaatgat tcatctaaag
     1081 tgacgaagga attggagaaa acggattata ttcatgtacc ggtaatggcg gaggatgttt
     1141 cctcagcggc cccgagcccg cggggctgtg cggatggtag ggatgccgac cctactgagg
     1201 agcagatggc agaaacagag agaaacgacg aggagcagtt cgaatgccag gaactgctcg
     1261 agtgccaggt gcaggtgggg gcccccgagg aggaggagga ggaggaggag gacgcgggcc
     1321 tggtggccga ggccgaggcc gtggctgccg gctggatgct cgatttcctc tgcctctctc
     1381 tttgccgagc tttccgcgac ggccgctccg aggacttccg caggacccgc aacagcgcag
     1441 aggctattat tcatggacta tccagtctaa cagcttgcca gttgagaacg atatacatat
     1501 gtcagttttt gacaagaatt gcagcaggaa aaacccttga tgcacagttt gaaaatgatg
     1561 aacgaattac acccttggaa tcagccctga tgatttgggg ttcaattgaa aaggaacatg
     1621 acaaacttca tgaagaaata cagaatttaa ttaaaattca ggctatagct gtttgtatgg
     1681 aaaatggcaa ctttaaagaa gcagaagaag tctttgaaag aatatttggt gatccaaatt
     1741 ctcatatgcc tttcaaaagc aaattgctta tgataatctc tcagaaagat acatttcatt
     1801 ccttttttca acacttcagc tacaaccaca tgatggagaa aattaagagt tatgtgaatt
     1861 atgtgctaag tgaaaaatca tcaacctttc taatgaaggc agcggcaaaa gtagtagaaa
     1921 gcaaaaggac aagaacaata acttctcaag ataaacctag tggtaatgat gttgaaatgg
     1981 aaactgaagc taatttggat acaagaaaaa ggtctcacaa gaatcttttc ttatctaagt
     2041 tgcaacatgg aacccagcaa caagacctta ataagaaaga aagaagagta ggaactcctc
     2101 aaagtacaaa aaagaaaaaa gaaagcagaa gagccactga aagcagaata cctgtttcaa
     2161 agagtcagcc ggtaactcct gaaaaacatc gagctagaaa aagacaggca tggctttggg
     2221 aagaagacaa gaatttgaga tctggcgtga ggaaatatgg agagggaaac tggtctaaaa
     2281 tactgttgca ttataaattc aacaaccgga caagtgtcat gttaaaagac agatggagga
     2341 ccatgaagaa actaaaactg atttcctcag acagcgaaga caagcttatg gtgtctaagg
     2401 gcgaagagct gattaaggag aacatgcaca tgaagctgta catggagggc accgtgaaca
     2461 accaccactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
     2521 tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacca
     2581 gcttcatgta cggcagcaga accttcatca accacaccca gggcatcccc gatttcttta
     2641 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg
     2701 tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga
     2761 tcagaggggt gaacttccca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg
     2821 aggccaacac cgagatgctg taccccgctg acggcggcct ggaaggcaga accgacatgg
     2881 ccctgaagct cgtgggcggg ggccacctga tctgcaactt caagaccaca tacagatcca
     2941 agaaacccgc taagaacctc aagatgcccg gcgtctacta tgtggaccac agactggaaa
     3001 gaatcaagga ggccgacaaa gagacctacg tcgagcagca cgaggtggct gtggccagat
     3061 actgcgacct ccctagcaaa ctggggcaca aacttaatta aggatccacc ggatctagat
     3121 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     3181 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     3241 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3301 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
     3361 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     3421 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     3481 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     3541 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     3601 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3661 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3721 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3781 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     3841 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3901 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     3961 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4021 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4081 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4141 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4201 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     4261 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     4321 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4381 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4441 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4501 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     4561 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4621 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4681 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4741 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4801 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4861 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4921 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4981 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     5041 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5101 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     5161 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5221 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5281 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     5341 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     5401 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     5461 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     5521 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     5581 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5641 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5701 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5761 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5821 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5881 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5941 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6001 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6061 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6121 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6181 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6241 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6301 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6361 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6421 ataaccgtat taccgccatg cat
//
