LOCUS       sequence-199212-        6498 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 6498)
  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 6498)
  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-EYFP-hTRF1
FEATURES             Location/Qualifiers
     source          1..6498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        49..352
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        353..556
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     507..527
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             595..1104
                     /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"
     regulatory      1117..1126
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1123..1125,1126..1128,1129..1839)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1168..1189)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1429..1448)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1776..1797
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3274..3395
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3311..3330)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3365..3384
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3402..3857)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3539..3560)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3751..3770
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3884..3988
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3990..4347
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3990..4181
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4013..4033)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4198..4333
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4382..5176
                     /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             4382..5176
                     /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(4436..4455)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5046..5065
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5364..5383)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5408..5455
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5784..6372
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6273..6292
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
       61 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      121 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      181 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      241 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      301 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      361 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      421 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      481 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      541 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctcgc caccatgaat
      601 ggagctatag gaggtgacct tttgctcaat tttcctgaca tgtcggtcct agagcgccaa
      661 agggctcacc tcaagtacct caatcccacc tttgattctc ctctcgccgg cttctttgcc
      721 gattcttcaa tgattaccgg cggcgagatg gacagctatc tttcgactgc cggtttgaat
      781 cttccgatga tgtacggtga gacgacggtg gaaggtgatt caagactctc aatttcgccg
      841 gaaacgacgc ttgggactgg aaatttcaag gcagcgaagt ttgatacaga gactaaggat
      901 tgtaatgagg cggcgaagaa gatgacgatg aacagagatg acctagtaga agaaggagaa
      961 gaagagaagt cgaaaataac agagcaaaac aatgggagca caaaaagcat caagaagatg
     1021 aaacacaaag ccaagaaaga agagaacaat ttctctaatg attcatctaa agtgacgaag
     1081 gaattggaga aaacggatta tattcatgga ccggtcgcca ccatggtgag caagggcgag
     1141 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     1201 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1261 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc
     1321 tacggcctgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     1381 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1441 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1501 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1561 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     1621 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1681 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
     1741 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1801 gccgccggga tcactctcgg catggacgag ctgtacaagt ccggactcag atctcgagct
     1861 caagcttcga attctgcagt cgacatggcg gaggatgttt cctcagcggc cccgagcccg
     1921 cggggctgtg cggatggtag ggatgccgac cctactgagg agcagatggc agaaacagag
     1981 agaaacgacg aggagcagtt cgaatgccag gaactgctcg agtgccaggt gcaggtgggg
     2041 gcccccgagg aggaggagga ggaggaggag gacgcgggcc tggtggccga ggccgaggcc
     2101 gtggctgccg gctggatgct cgatttcctc tgcctctctc tttgccgagc tttccgcgac
     2161 ggccgctccg aggacttccg caggacccgc aacagcgcag aggctattat tcatggacta
     2221 tccagtctaa cagcttgcca gttgagaacg atatacatat gtcagttttt gacaagaatt
     2281 gcagcaggaa aaacccttga tgcacagttt gaaaatgatg aacgaattac acccttggaa
     2341 tcagccctga tgatttgggg ttcaattgaa aaggaacatg acaaacttca tgaagaaata
     2401 cagaatttaa ttaaaattca ggctatagct gtttgtatgg aaaatggcaa ctttaaagaa
     2461 gcagaagaag tctttgaaag aatatttggt gatccaaatt ctcatatgcc tttcaaaagc
     2521 aaattgctta tgataatctc tcagaaagat acatttcatt ccttttttca acacttcagc
     2581 tacaaccaca tgatggagaa aattaagagt tatgtgaatt atgtgctaag tgaaaaatca
     2641 tcaacctttc taatgaaggc agcggcaaaa gtagtagaaa gcaaaaggac aagaacaata
     2701 acttctcaag ataaacctag tggtaatgat gttgaaatgg aaactgaagc taatttggat
     2761 acaagaaaaa ggtctcacaa gaatcttttc ttatctaagt tgcaacatgg aacccagcaa
     2821 caagacctta ataagaaaga aagaagagta ggaactcctc aaagtacaaa aaagaaaaaa
     2881 gaaagcagaa gagccactga aagcagaata cctgtttcaa agagtcagcc ggtaactcct
     2941 gaaaaacatc gagctagaaa aagacaggca tggctttggg aagaagacaa gaatttgaga
     3001 tctggcgtga ggaaatatgg agagggaaac tggtctaaaa tactgttgca ttataaattc
     3061 aacaaccgga caagtgtcat gttaaaagac agatggagga ccatgaagaa actaaaactg
     3121 atttcctcag acagcgaaga ctgaggatcc accggatcta gataactgat cataatcagc
     3181 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     3241 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     3301 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3361 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     3421 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3481 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3541 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3601 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     3661 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3721 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     3781 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3841 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3901 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3961 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     4021 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4081 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4141 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4201 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4261 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4321 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     4381 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4441 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4501 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4561 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4621 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4681 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4741 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4801 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4861 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     4921 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4981 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5041 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5101 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5161 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5221 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5281 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5341 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     5401 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     5461 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     5521 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     5581 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     5641 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5701 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5761 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5821 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5881 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5941 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6001 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6061 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6121 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6181 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6241 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6301 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6361 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6421 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6481 atgcattagt tattaata
//
