LOCUS       sequence-196668-        6721 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 6721)
  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 6721)
  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-hTRF2
FEATURES             Location/Qualifiers
     source          1..6721
                     /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    3509..3630
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3546..3565)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3600..3619
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3637..4092)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3774..3795)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3986..4005
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4119..4223
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4225..4582
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4225..4416
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4248..4268)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4433..4568
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4617..5411
                     /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             4617..5411
                     /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(4671..4690)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5281..5300
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5599..5618)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5643..5690
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6019..6607
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6508..6527
                     /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 ggcgggagga ggcgggagta gcgacggcag cgggcgggca
     1921 gctggcaggc gggcgtcccg cagtagcggg cgggcccggc gggggcgcca cgagccgggg
     1981 ctggggggcc cggcggagcg cggcgcgggg gaggcacggc tggaagaggc agtcaatcgc
     2041 tgggtgctca agttctactt ccacgaggcg ctgcgggcct ttcggggtag ccggtacggg
     2101 gacttcagac agatccggga catcatgcag gctttgcttg tcaggccctt ggggaaggag
     2161 cacaccgtgt cccgattgct gcgggttatg cagtgtctgt cgcggattga agaaggggaa
     2221 aatttagact gttcctttga tatggaggct gagctcacac cactggaatc agctatcaat
     2281 gtgctggaga tgattaaaac ggaatttaca ctgacagaag cagtggtcga atccagtaga
     2341 aaactggtca aggaagctgc tgtcattatt tgtatcaaaa acaaagaatt tgaaaaggct
     2401 tcaaaaattt tgaaaaaaca tatgtccaag gaccccacaa ctcagaagct gagaaatgat
     2461 ctcctgaata ttattcgaga aaagaacttg gcccatcctg ttatccagaa cttttcatat
     2521 gagaccttcc agcagaagat gctgcgcttc ctggagagcc acctggatga cgccgagccc
     2581 tacctcctca cgatggccaa aaaggctttg aaatctgagt ccgctgcctc aagtacaggg
     2641 aaggaagata aacagccagc accagggcct gtggaaaagc cacccagaga acccgcaagg
     2701 cagctacgga atcctccaac caccattgga atgatgactc tgaaagcagc tttcaagact
     2761 ctgtctggtg cacaggattc tgaggcagcc tttgcaaaac tggaccagaa ggatctggtt
     2821 cttcctactc aagctctccc agcatcacca gccctcaaaa acaagagacc cagaaaagat
     2881 gaaaacgaaa gttcagcccc ggctgacggt gagggtggct cggaactgca gcccaagaac
     2941 aagcgcatga caataagcag attggtcttg gaggaggaca gccagagtac tgagcccagc
     3001 gcaggcctca actcctccca ggaggccgct tcagcgccac catccaagcc caccgttctc
     3061 aaccaacccc tccctggaga gaagaatccc aaagtaccca aaggcaagtg gaacagctct
     3121 aatggggttg aagaaaagga gacttgggtg gaagaggatg aactgtttca agttcaggca
     3181 ccagatgaag acagtacaac caatataaca aaaaagcaga agtggactgt agaagaaagc
     3241 gagtgggtca aggctggagt gcagaaatat ggggaaggaa actgggctgc catttctaaa
     3301 aattacccat ttgttaaccg aacagctgtg atgattaagg atcgctggcg gaccatgaaa
     3361 agacttggca tgaactgagg gatccaccgg atctagataa ctgatcataa tcagccatac
     3421 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     3481 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     3541 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3601 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     3661 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3721 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3781 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3841 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3901 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3961 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4021 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4081 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4141 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4201 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     4261 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4321 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4381 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4441 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4501 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4561 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     4621 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4681 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4741 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     4801 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4861 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4921 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4981 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5041 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5101 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     5161 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5221 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5281 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5341 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5401 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5461 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5521 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5581 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     5641 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     5701 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     5761 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     5821 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     5881 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5941 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6001 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6061 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6121 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     6181 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6241 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6301 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6361 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6421 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6481 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6541 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6601 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6661 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     6721 t
//
