LOCUS       sequence-199232-        6739 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 6739)
  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 6739)
  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-hTRF2
FEATURES             Location/Qualifiers
     source          1..6739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        58..361
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        362..565
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     516..536
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             604..1113
                     /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      1126..1135
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1132..1134,1135..1137,1138..1848)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1177..1198)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1438..1457)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1785..1806
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3524..3645
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3561..3580)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3615..3634
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3652..4107)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3789..3810)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4001..4020
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4134..4238
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4240..4597
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4240..4431
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4263..4283)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4448..4583
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4632..5426
                     /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             4632..5426
                     /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(4686..4705)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5296..5315
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5614..5633)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5658..5705
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6034..6622
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6523..6542
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
       61 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      121 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      181 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      241 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      301 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      361 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      421 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      481 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      541 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgctcgcc
      601 accatgaatg gagctatagg aggtgacctt ttgctcaatt ttcctgacat gtcggtccta
      661 gagcgccaaa gggctcacct caagtacctc aatcccacct ttgattctcc tctcgccggc
      721 ttctttgccg attcttcaat gattaccggc ggcgagatgg acagctatct ttcgactgcc
      781 ggtttgaatc ttccgatgat gtacggtgag acgacggtgg aaggtgattc aagactctca
      841 atttcgccgg aaacgacgct tgggactgga aatttcaagg cagcgaagtt tgatacagag
      901 actaaggatt gtaatgaggc ggcgaagaag atgacgatga acagagatga cctagtagaa
      961 gaaggagaag aagagaagtc gaaaataaca gagcaaaaca atgggagcac aaaaagcatc
     1021 aagaagatga aacacaaagc caagaaagaa gagaacaatt tctctaatga ttcatctaaa
     1081 gtgacgaagg aattggagaa aacggattat attcatggac cggtcgccac catggtgagc
     1141 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1201 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     1261 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1321 accttcggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     1381 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1441 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1501 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1561 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     1621 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     1681 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1741 taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1801 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc cggactcaga
     1861 tctcgagctc aagcttcgaa ttctgcagtc gacatggcgg gaggaggcgg gagtagcgac
     1921 ggcagcgggc gggcagctgg caggcgggcg tcccgcagta gcgggcgggc ccggcggggg
     1981 cgccacgagc cggggctggg gggcccggcg gagcgcggcg cgggggaggc acggctggaa
     2041 gaggcagtca atcgctgggt gctcaagttc tacttccacg aggcgctgcg ggcctttcgg
     2101 ggtagccggt acggggactt cagacagatc cgggacatca tgcaggcttt gcttgtcagg
     2161 cccttgggga aggagcacac cgtgtcccga ttgctgcggg ttatgcagtg tctgtcgcgg
     2221 attgaagaag gggaaaattt agactgttcc tttgatatgg aggctgagct cacaccactg
     2281 gaatcagcta tcaatgtgct ggagatgatt aaaacggaat ttacactgac agaagcagtg
     2341 gtcgaatcca gtagaaaact ggtcaaggaa gctgctgtca ttatttgtat caaaaacaaa
     2401 gaatttgaaa aggcttcaaa aattttgaaa aaacatatgt ccaaggaccc cacaactcag
     2461 aagctgagaa atgatctcct gaatattatt cgagaaaaga acttggccca tcctgttatc
     2521 cagaactttt catatgagac cttccagcag aagatgctgc gcttcctgga gagccacctg
     2581 gatgacgccg agccctacct cctcacgatg gccaaaaagg ctttgaaatc tgagtccgct
     2641 gcctcaagta cagggaagga agataaacag ccagcaccag ggcctgtgga aaagccaccc
     2701 agagaacccg caaggcagct acggaatcct ccaaccacca ttggaatgat gactctgaaa
     2761 gcagctttca agactctgtc tggtgcacag gattctgagg cagcctttgc aaaactggac
     2821 cagaaggatc tggttcttcc tactcaagct ctcccagcat caccagccct caaaaacaag
     2881 agacccagaa aagatgaaaa cgaaagttca gccccggctg acggtgaggg tggctcggaa
     2941 ctgcagccca agaacaagcg catgacaata agcagattgg tcttggagga ggacagccag
     3001 agtactgagc ccagcgcagg cctcaactcc tcccaggagg ccgcttcagc gccaccatcc
     3061 aagcccaccg ttctcaacca acccctccct ggagagaaga atcccaaagt acccaaaggc
     3121 aagtggaaca gctctaatgg ggttgaagaa aaggagactt gggtggaaga ggatgaactg
     3181 tttcaagttc aggcaccaga tgaagacagt acaaccaata taacaaaaaa gcagaagtgg
     3241 actgtagaag aaagcgagtg ggtcaaggct ggagtgcaga aatatgggga aggaaactgg
     3301 gctgccattt ctaaaaatta cccatttgtt aaccgaacag ctgtgatgat taaggatcgc
     3361 tggcggacca tgaaaagact tggcatgaac tgagggatcc accggatcta gataactgat
     3421 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     3481 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3541 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3601 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     3661 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3721 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3781 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3841 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3901 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3961 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     4021 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     4081 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4141 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4201 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     4261 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4321 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4381 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4441 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4501 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4561 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     4621 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4681 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4741 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4801 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4861 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4921 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4981 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5041 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5101 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     5161 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5221 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5281 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5341 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5401 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5461 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5521 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5581 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     5641 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     5701 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     5761 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     5821 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     5881 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5941 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6001 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6061 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6121 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6181 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6241 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6301 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6361 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6421 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6481 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6541 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6601 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6661 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6721 tattaccgcc atgcattag
//
