LOCUS       sequence-199226-        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        59..362
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        363..566
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     517..537
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             605..1114
                     /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             1133..1831
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    3507..3628
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3544..3563)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3598..3617
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3635..4090)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3772..3793)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3984..4003
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4117..4221
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4223..4580
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4223..4414
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4246..4266)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4431..4566
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4615..5409
                     /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             4615..5409
                     /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(4669..4688)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5279..5298
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5597..5616)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5641..5688
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6017..6605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6506..6525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
       61 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      121 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      181 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      241 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      301 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      361 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      421 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      481 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      541 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctcgc
      601 caccatgaat ggagctatag gaggtgacct tttgctcaat tttcctgaca tgtcggtcct
      661 agagcgccaa agggctcacc tcaagtacct caatcccacc tttgattctc ctctcgccgg
      721 cttctttgcc gattcttcaa tgattaccgg cggcgagatg gacagctatc tttcgactgc
      781 cggtttgaat cttccgatga tgtacggtga gacgacggtg gaaggtgatt caagactctc
      841 aatttcgccg gaaacgacgc ttgggactgg aaatttcaag gcagcgaagt ttgatacaga
      901 gactaaggat tgtaatgagg cggcgaagaa gatgacgatg aacagagatg acctagtaga
      961 agaaggagaa gaagagaagt cgaaaataac agagcaaaac aatgggagca caaaaagcat
     1021 caagaagatg aaacacaaag ccaagaaaga agagaacaat ttctctaatg attcatctaa
     1081 agtgacgaag gaattggaga aaacggatta tattcatgga ccggtcgcca ccatgagcga
     1141 gctgattaag gagaacatgc acatgaagct gtacatggag ggcaccgtgg acaaccatca
     1201 cttcaagtgc acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgagaat
     1261 caaggtggtc gagggcggcc ctctcccctt cgccttcgac atcctggcta ctagcttcct
     1321 ctacggcagc aagaccttca tcaaccacac ccagggcatc cccgacttct tcaagcagtc
     1381 cttccctgag ggcttcacat gggagagagt caccacatac gaagacgggg gcgtgctgac
     1441 cgctacccag gacaccagcc tccaggacgg ctgcctcatc tacaacgtca agatcagagg
     1501 ggtgaacttc acatccaacg gccctgtgat gcagaagaaa acactcggct gggaggcctt
     1561 caccgagacg ctgtaccccg ctgacggcgg cctggaaggc agaaacgaca tggccctgaa
     1621 gctcgtgggc gggagccatc tgatcgcaaa catcaagacc acatatagat ccaagaaacc
     1681 cgctaagaac ctcaagatgc ctggcgtcta ctatgtggac tacagactgg aaagaatcaa
     1741 ggaggccaac aacgagacct acgtcgagca gcacgaggtg gcagtggcca gatactgcga
     1801 cctccctagc aaactggggc acaagcttaa ttccggactc agatctcgag ctcaagcttc
     1861 gaattctgca gtcgacatgg cgggaggagg cgggagtagc gacggcagcg ggcgggcagc
     1921 tggcaggcgg gcgtcccgca gtagcgggcg ggcccggcgg gggcgccacg agccggggct
     1981 ggggggcccg gcggagcgcg gcgcggggga ggcacggctg gaagaggcag tcaatcgctg
     2041 ggtgctcaag ttctacttcc acgaggcgct gcgggccttt cggggtagcc ggtacgggga
     2101 cttcagacag atccgggaca tcatgcaggc tttgcttgtc aggcccttgg ggaaggagca
     2161 caccgtgtcc cgattgctgc gggttatgca gtgtctgtcg cggattgaag aaggggaaaa
     2221 tttagactgt tcctttgata tggaggctga gctcacacca ctggaatcag ctatcaatgt
     2281 gctggagatg attaaaacgg aatttacact gacagaagca gtggtcgaat ccagtagaaa
     2341 actggtcaag gaagctgctg tcattatttg tatcaaaaac aaagaatttg aaaaggcttc
     2401 aaaaattttg aaaaaacata tgtccaagga ccccacaact cagaagctga gaaatgatct
     2461 cctgaatatt attcgagaaa agaacttggc ccatcctgtt atccagaact tttcatatga
     2521 gaccttccag cagaagatgc tgcgcttcct ggagagccac ctggatgacg ccgagcccta
     2581 cctcctcacg atggccaaaa aggctttgaa atctgagtcc gctgcctcaa gtacagggaa
     2641 ggaagataaa cagccagcac cagggcctgt ggaaaagcca cccagagaac ccgcaaggca
     2701 gctacggaat cctccaacca ccattggaat gatgactctg aaagcagctt tcaagactct
     2761 gtctggtgca caggattctg aggcagcctt tgcaaaactg gaccagaagg atctggttct
     2821 tcctactcaa gctctcccag catcaccagc cctcaaaaac aagagaccca gaaaagatga
     2881 aaacgaaagt tcagccccgg ctgacggtga gggtggctcg gaactgcagc ccaagaacaa
     2941 gcgcatgaca ataagcagat tggtcttgga ggaggacagc cagagtactg agcccagcgc
     3001 aggcctcaac tcctcccagg aggccgcttc agcgccacca tccaagccca ccgttctcaa
     3061 ccaacccctc cctggagaga agaatcccaa agtacccaaa ggcaagtgga acagctctaa
     3121 tggggttgaa gaaaaggaga cttgggtgga agaggatgaa ctgtttcaag ttcaggcacc
     3181 agatgaagac agtacaacca atataacaaa aaagcagaag tggactgtag aagaaagcga
     3241 gtgggtcaag gctggagtgc agaaatatgg ggaaggaaac tgggctgcca tttctaaaaa
     3301 ttacccattt gttaaccgaa cagctgtgat gattaaggat cgctggcgga ccatgaaaag
     3361 acttggcatg aactgaggga tccaccggat ctagataact gatcataatc agccatacca
     3421 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3481 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3541 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3601 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     3661 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3721 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3781 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3841 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3901 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3961 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4021 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4081 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4141 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4201 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4261 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4321 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4381 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4441 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4501 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4561 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4621 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4681 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4741 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     4801 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4861 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4921 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4981 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5041 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5101 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     5161 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5221 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5281 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5341 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5401 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5461 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5521 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5581 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     5641 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     5701 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     5761 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     5821 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     5881 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5941 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6001 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6061 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6601 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6661 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt
     6721 a
//
