LOCUS       sequence-196695-        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        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             2628..3341
                     /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    3471..3592
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3508..3527)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3562..3581
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3599..4054)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3736..3757)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3948..3967
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4081..4185
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4187..4544
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4187..4378
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4210..4230)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4395..4530
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4579..5373
                     /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             4579..5373
                     /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(4633..4652)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5243..5262
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5561..5580)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5605..5652
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5981..6569
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6470..6489
                     /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 ggaggaggcg
     1141 ggagtagcga cggcagcggg cgggcagctg gcaggcgggc gtcccgcagt agcgggcggg
     1201 cccggcgggg gcgccacgag ccggggctgg ggggcccggc ggagcgcggc gcgggggagg
     1261 cacggctgga agaggcagtc aatcgctggg tgctcaagtt ctacttccac gaggcgctgc
     1321 gggcctttcg gggtagccgg tacggggact tcagacagat ccgggacatc atgcaggctt
     1381 tgcttgtcag gcccttgggg aaggagcaca ccgtgtcccg attgctgcgg gttatgcagt
     1441 gtctgtcgcg gattgaagaa ggggaaaatt tagactgttc ctttgatatg gaggctgagc
     1501 tcacaccact ggaatcagct atcaatgtgc tggagatgat taaaacggaa tttacactga
     1561 cagaagcagt ggtcgaatcc agtagaaaac tggtcaagga agctgctgtc attatttgta
     1621 tcaaaaacaa agaatttgaa aaggcttcaa aaattttgaa aaaacatatg tccaaggacc
     1681 ccacaactca gaagctgaga aatgatctcc tgaatattat tcgagaaaag aacttggccc
     1741 atcctgttat ccagaacttt tcatatgaga ccttccagca gaagatgctg cgcttcctgg
     1801 agagccacct ggatgacgcc gagccctacc tcctcacgat ggccaaaaag gctttgaaat
     1861 ctgagtccgc tgcctcaagt acagggaagg aagataaaca gccagcacca gggcctgtgg
     1921 aaaagccacc cagagaaccc gcaaggcagc tacggaatcc tccaaccacc attggaatga
     1981 tgactctgaa agcagctttc aagactctgt ctggtgcaca ggattctgag gcagcctttg
     2041 caaaactgga ccagaaggat ctggttcttc ctactcaagc tctcccagca tcaccagccc
     2101 tcaaaaacaa gagacccaga aaagatgaaa acgaaagttc agccccggct gacggtgagg
     2161 gtggctcgga actgcagccc aagaacaagc gcatgacaat aagcagattg gtcttggagg
     2221 aggacagcca gagtactgag cccagcgcag gcctcaactc ctcccaggag gccgcttcag
     2281 cgccaccatc caagcccacc gttctcaacc aacccctccc tggagagaag aatcccaaag
     2341 tacccaaagg caagtggaac agctctaatg gggttgaaga aaaggagact tgggtggaag
     2401 aggatgaact gtttcaagtt caggcaccag atgaagacag tacaaccaat ataacaaaaa
     2461 agcagaagtg gactgtagaa gaaagcgagt gggtcaaggc tggagtgcag aaatatgggg
     2521 aaggaaactg ggctgccatt tctaaaaatt acccatttgt taaccgaaca gctgtgatga
     2581 ttaaggatcg ctggcggacc atgaaaagac ttggcatgaa gaagcttatg gtgtctaagg
     2641 gcgaagagct gattaaggag aacatgcaca tgaagctgta catggagggc accgtgaaca
     2701 accaccactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
     2761 tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacca
     2821 gcttcatgta cggcagcaga accttcatca accacaccca gggcatcccc gatttcttta
     2881 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg
     2941 tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga
     3001 tcagaggggt gaacttccca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg
     3061 aggccaacac cgagatgctg taccccgctg acggcggcct ggaaggcaga accgacatgg
     3121 ccctgaagct cgtgggcggg ggccacctga tctgcaactt caagaccaca tacagatcca
     3181 agaaacccgc taagaacctc aagatgcccg gcgtctacta tgtggaccac agactggaaa
     3241 gaatcaagga ggccgacaaa gagacctacg tcgagcagca cgaggtggct gtggccagat
     3301 actgcgacct ccctagcaaa ctggggcaca aacttaatta aggatccacc ggatctagat
     3361 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     3421 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     3481 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3541 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
     3601 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     3661 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     3721 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     3781 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     3841 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3901 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3961 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     4021 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     4081 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4141 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     4201 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4261 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4321 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4381 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4441 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     4501 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     4561 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4621 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4681 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4741 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     4801 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4861 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4921 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4981 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     5041 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     5101 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     5161 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     5221 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     5281 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5341 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     5401 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5461 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5521 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     5581 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     5641 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     5701 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     5761 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     5821 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5881 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5941 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6001 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6061 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6121 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6181 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6241 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6301 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6361 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6421 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6481 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6541 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6601 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6661 ataaccgtat taccgccatg cat
//
