LOCUS       sequence-199277-        6683 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 6683)
  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 6683)
  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-hTRF2-tagRFP-T
FEATURES             Location/Qualifiers
     source          1..6683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        56..359
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        360..563
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     514..534
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             598..1107
                     /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             2623..3336
                     /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    3466..3587
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3503..3522)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3557..3576
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3594..4049)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3731..3752)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3943..3962
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4076..4180
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4182..4539
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4182..4373
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4205..4225)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4390..4525
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4574..5368
                     /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             4574..5368
                     /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(4628..4647)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5238..5257
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5556..5575)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5600..5647
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5976..6564
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6465..6484
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
       61 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      121 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      181 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      241 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      301 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      361 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      421 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      481 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      541 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag cgccaccatg
      601 aatggagcta taggaggtga ccttttgctc aattttcctg acatgtcggt cctagagcgc
      661 caaagggctc acctcaagta cctcaatccc acctttgatt ctcctctcgc cggcttcttt
      721 gccgattctt caatgattac cggcggcgag atggacagct atctttcgac tgccggtttg
      781 aatcttccga tgatgtacgg tgagacgacg gtggaaggtg attcaagact ctcaatttcg
      841 ccggaaacga cgcttgggac tggaaatttc aaggcagcga agtttgatac agagactaag
      901 gattgtaatg aggcggcgaa gaagatgacg atgaacagag atgacctagt agaagaagga
      961 gaagaagaga agtcgaaaat aacagagcaa aacaatggga gcacaaaaag catcaagaag
     1021 atgaaacaca aagccaagaa agaagagaac aatttctcta atgattcatc taaagtgacg
     1081 aaggaattgg agaaaacgga ttatattcat gtaccggtaa tggcgggagg aggcgggagt
     1141 agcgacggca gcgggcgggc agctggcagg cgggcgtccc gcagtagcgg gcgggcccgg
     1201 cgggggcgcc acgagccggg gctggggggc ccggcggagc gcggcgcggg ggaggcacgg
     1261 ctggaagagg cagtcaatcg ctgggtgctc aagttctact tccacgaggc gctgcgggcc
     1321 tttcggggta gccggtacgg ggacttcaga cagatccggg acatcatgca ggctttgctt
     1381 gtcaggccct tggggaagga gcacaccgtg tcccgattgc tgcgggttat gcagtgtctg
     1441 tcgcggattg aagaagggga aaatttagac tgttcctttg atatggaggc tgagctcaca
     1501 ccactggaat cagctatcaa tgtgctggag atgattaaaa cggaatttac actgacagaa
     1561 gcagtggtcg aatccagtag aaaactggtc aaggaagctg ctgtcattat ttgtatcaaa
     1621 aacaaagaat ttgaaaaggc ttcaaaaatt ttgaaaaaac atatgtccaa ggaccccaca
     1681 actcagaagc tgagaaatga tctcctgaat attattcgag aaaagaactt ggcccatcct
     1741 gttatccaga acttttcata tgagaccttc cagcagaaga tgctgcgctt cctggagagc
     1801 cacctggatg acgccgagcc ctacctcctc acgatggcca aaaaggcttt gaaatctgag
     1861 tccgctgcct caagtacagg gaaggaagat aaacagccag caccagggcc tgtggaaaag
     1921 ccacccagag aacccgcaag gcagctacgg aatcctccaa ccaccattgg aatgatgact
     1981 ctgaaagcag ctttcaagac tctgtctggt gcacaggatt ctgaggcagc ctttgcaaaa
     2041 ctggaccaga aggatctggt tcttcctact caagctctcc cagcatcacc agccctcaaa
     2101 aacaagagac ccagaaaaga tgaaaacgaa agttcagccc cggctgacgg tgagggtggc
     2161 tcggaactgc agcccaagaa caagcgcatg acaataagca gattggtctt ggaggaggac
     2221 agccagagta ctgagcccag cgcaggcctc aactcctccc aggaggccgc ttcagcgcca
     2281 ccatccaagc ccaccgttct caaccaaccc ctccctggag agaagaatcc caaagtaccc
     2341 aaaggcaagt ggaacagctc taatggggtt gaagaaaagg agacttgggt ggaagaggat
     2401 gaactgtttc aagttcaggc accagatgaa gacagtacaa ccaatataac aaaaaagcag
     2461 aagtggactg tagaagaaag cgagtgggtc aaggctggag tgcagaaata tggggaagga
     2521 aactgggctg ccatttctaa aaattaccca tttgttaacc gaacagctgt gatgattaag
     2581 gatcgctggc ggaccatgaa aagacttggc atgaagaagc ttatggtgtc taagggcgaa
     2641 gagctgatta aggagaacat gcacatgaag ctgtacatgg agggcaccgt gaacaaccac
     2701 cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga
     2761 atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc taccagcttc
     2821 atgtacggca gcagaacctt catcaaccac acccagggca tccccgactt ctttaagcag
     2881 tccttccctg agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg
     2941 accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga
     3001 ggggtgaact tcccatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc
     3061 aacaccgaga tgctgtaccc cgctgacggc ggcctggaag gcagaaccga catggccctg
     3121 aagctcgtgg gcgggggcca cctgatctgc aacttcaaga ccacatacag atccaagaaa
     3181 cccgctaaga acctcaagat gcccggcgtc tactatgtgg accacagact ggaaagaatc
     3241 aaggaggccg acaaagagac ctacgtcgag cagcacgagg tggctgtggc cagatactgc
     3301 gacctcccta gcaaactggg gcacaaactt aattaaggat ccaccggatc tagataactg
     3361 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3421 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3481 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3541 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt
     3601 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     3661 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3721 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3781 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3841 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3901 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3961 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     4021 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     4081 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4141 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     4201 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4261 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4321 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4381 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4441 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4501 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     4561 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4621 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4681 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4741 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4801 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4861 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4921 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4981 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5041 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5101 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5161 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5221 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5281 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5341 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5401 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5461 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5521 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     5581 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     5641 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     5701 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5761 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     5821 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5881 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5941 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6001 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6061 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6121 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6181 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6241 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6301 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6361 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6421 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6481 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6541 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6601 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6661 cgtattaccg ccatgcatta gtt
//
