LOCUS       sequence-196659-        7006 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BLInCR 'Localizer' construct that marks the nuclear lamina 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 7006)
  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 7006)
  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-hLaminB1
FEATURES             Location/Qualifiers
     source          1..7006
                     /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"
     regulatory      1129..1138
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1135..1137,1138..1140,1141..1851)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1180..1201)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1441..1460)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1788..1809
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3794..3915
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3831..3850)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3885..3904
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3922..4377)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4059..4080)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4271..4290
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4404..4508
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4510..4867
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4510..4701
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4533..4553)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4718..4853
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4902..5696
                     /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             4902..5696
                     /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(4956..4975)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5566..5585
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5884..5903)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5928..5975
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6304..6892
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6793..6812
                     /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 caccatggtg
     1141 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1201 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     1261 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1321 accaccttcg gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
     1381 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1441 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1501 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1561 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     1621 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     1681 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1741 agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1801 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtccggactc
     1861 agatctcgag ctcaagcttc gaattctgca gtcgacatgg cgactgcgac ccccgtgccg
     1921 ccgcggatgg gcagccgcgc tggcggcccc accacgccgc tgagccccac gcgcctgtcg
     1981 cggctccagg agaaggagga gctgcgcgag ctcaatgacc ggctggcggt gtacatcgac
     2041 aaggtgcgca gcctggagac ggagaacagc gcgctgcagc tgcaggtgac ggagcgcgag
     2101 gaggtgcgcg gccgtgagct caccggcctc aaggcgctct acgagaccga gctggccgac
     2161 gcgcgacgcg cgctcgacga cacggcccgc gagcgcgcca agctgcagat cgagctgggc
     2221 aagtgcaagg cggaacacga ccagctgctc ctcaactatg ctaagaagga atctgatctt
     2281 aatggcgccc agatcaagct tcgagaatat gaagcagcac tgaattcgaa agatgcagct
     2341 cttgctactg cacttggtga caaaaaaagt ttagagggag atttggagga tctgaaggat
     2401 cagattgccc agttggaagc ctccttagct gcagccaaaa aacagttagc agatgaaact
     2461 ttacttaaag tagatttgga gaatcgttgt cagagcctta ctgaggactt ggagtttcgc
     2521 aaaagcatgt atgaagagga gattaacgag accagaagga agcatgaaac gcgcttggta
     2581 gaggtggatt ctgggcgtca aattgagtat gagtacaagc tggcgcaagc ccttcatgag
     2641 atgagagagc aacatgatgc ccaagtgagg ctgtataagg aggagctgga gcagacttac
     2701 catgccaaac ttgagaatgc cagactgtca tcagagatga atacttctac tgtcaacagt
     2761 gccagggaag aactgatgga aagccgcatg agaattgaga gcctttcatc ccagctttct
     2821 aatctacaga aagagtctag agcatgtttg gaaaggattc aagaattaga ggacttgctt
     2881 gctaaagaaa aagacaactc tcgtcgcatg ctgacagaca aagagagaga gatggcggaa
     2941 ataagggatc aaatgcagca acagctgaat gactatgaac agcttcttga tgtaaagtta
     3001 gccctggaca tggaaatcag tgcttacagg aaactcttag aaggcgaaga agagaggttg
     3061 aagctgtctc caagcccttc ttcccgtgtg acagtatccc gagcatcctc aagtcgtagt
     3121 gtacgtacaa ctagaggaaa gcggaagagg gttgatgtgg aagaatcaga ggcgagtagt
     3181 agtgttagca tctctcattc cgcctcagcc actggaaatg tttgcatcga agaaattgat
     3241 gttgatggga aatttatccg cttgaagaac acttctgaac aggatcaacc aatgggaggc
     3301 tgggagatga tcagaaaaat tggagacaca tcagtcagtt ataaatatac ctcaagatat
     3361 gtgctgaagg caggccagac tgttacaatt tgggctgcaa acgctggtgt cacagccagc
     3421 cccccaactg acctcatctg gaagaaccag aactcgtggg gcactggcga agatgtgaag
     3481 gttatattga aaaattctca gggagaggag gttgctcaaa gaagtacagt ctttaaaaca
     3541 accatacctg aagaagagga ggaggaggaa gaagcagctg gagtggttgt tgaggaagaa
     3601 cttttccacc agcagggaac cccaagagca tccaatagaa gctgtgcaat tatgtaaggg
     3661 cccgggatcc accggatcta gataactgat cataatcagc cataccacat ttgtagaggt
     3721 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     3781 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3841 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3901 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     3961 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     4021 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     4081 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     4141 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     4201 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     4261 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     4321 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     4381 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4441 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4501 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     4561 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4621 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4681 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4741 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4801 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4861 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4921 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4981 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5041 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     5101 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5161 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5221 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5281 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5341 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5401 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     5461 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5521 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5581 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5641 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5701 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5761 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5821 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     5881 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     5941 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     6001 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     6061 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     6121 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     6181 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6241 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6301 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6361 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6421 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     6481 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6541 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6601 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6661 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6721 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6781 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6841 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6901 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6961 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
