LOCUS       sequence-196656-        6988 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 6988)
  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 6988)
  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-hLaminB1
FEATURES             Location/Qualifiers
     source          1..6988
                     /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    3776..3897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3813..3832)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3867..3886
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3904..4359)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4041..4062)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4253..4272
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4386..4490
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4492..4849
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4492..4683
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4515..4535)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4700..4835
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4884..5678
                     /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             4884..5678
                     /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(4938..4957)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5548..5567
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5866..5885)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5910..5957
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6286..6874
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6775..6794
                     /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 ggcgactgcg acccccgtgc cgccgcggat gggcagccgc
     1921 gctggcggcc ccaccacgcc gctgagcccc acgcgcctgt cgcggctcca ggagaaggag
     1981 gagctgcgcg agctcaatga ccggctggcg gtgtacatcg acaaggtgcg cagcctggag
     2041 acggagaaca gcgcgctgca gctgcaggtg acggagcgcg aggaggtgcg cggccgtgag
     2101 ctcaccggcc tcaaggcgct ctacgagacc gagctggccg acgcgcgacg cgcgctcgac
     2161 gacacggccc gcgagcgcgc caagctgcag atcgagctgg gcaagtgcaa ggcggaacac
     2221 gaccagctgc tcctcaacta tgctaagaag gaatctgatc ttaatggcgc ccagatcaag
     2281 cttcgagaat atgaagcagc actgaattcg aaagatgcag ctcttgctac tgcacttggt
     2341 gacaaaaaaa gtttagaggg agatttggag gatctgaagg atcagattgc ccagttggaa
     2401 gcctccttag ctgcagccaa aaaacagtta gcagatgaaa ctttacttaa agtagatttg
     2461 gagaatcgtt gtcagagcct tactgaggac ttggagtttc gcaaaagcat gtatgaagag
     2521 gagattaacg agaccagaag gaagcatgaa acgcgcttgg tagaggtgga ttctgggcgt
     2581 caaattgagt atgagtacaa gctggcgcaa gcccttcatg agatgagaga gcaacatgat
     2641 gcccaagtga ggctgtataa ggaggagctg gagcagactt accatgccaa acttgagaat
     2701 gccagactgt catcagagat gaatacttct actgtcaaca gtgccaggga agaactgatg
     2761 gaaagccgca tgagaattga gagcctttca tcccagcttt ctaatctaca gaaagagtct
     2821 agagcatgtt tggaaaggat tcaagaatta gaggacttgc ttgctaaaga aaaagacaac
     2881 tctcgtcgca tgctgacaga caaagagaga gagatggcgg aaataaggga tcaaatgcag
     2941 caacagctga atgactatga acagcttctt gatgtaaagt tagccctgga catggaaatc
     3001 agtgcttaca ggaaactctt agaaggcgaa gaagagaggt tgaagctgtc tccaagccct
     3061 tcttcccgtg tgacagtatc ccgagcatcc tcaagtcgta gtgtacgtac aactagagga
     3121 aagcggaaga gggttgatgt ggaagaatca gaggcgagta gtagtgttag catctctcat
     3181 tccgcctcag ccactggaaa tgtttgcatc gaagaaattg atgttgatgg gaaatttatc
     3241 cgcttgaaga acacttctga acaggatcaa ccaatgggag gctgggagat gatcagaaaa
     3301 attggagaca catcagtcag ttataaatat acctcaagat atgtgctgaa ggcaggccag
     3361 actgttacaa tttgggctgc aaacgctggt gtcacagcca gccccccaac tgacctcatc
     3421 tggaagaacc agaactcgtg gggcactggc gaagatgtga aggttatatt gaaaaattct
     3481 cagggagagg aggttgctca aagaagtaca gtctttaaaa caaccatacc tgaagaagag
     3541 gaggaggagg aagaagcagc tggagtggtt gttgaggaag aacttttcca ccagcaggga
     3601 accccaagag catccaatag aagctgtgca attatgtaag ggcccgggat ccaccggatc
     3661 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3721 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3781 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3841 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     3901 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3961 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     4021 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     4081 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     4141 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     4201 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     4261 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     4321 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     4381 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4441 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     4501 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4561 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4621 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4681 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4741 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4801 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     4861 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4921 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4981 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5041 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     5101 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5161 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5221 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5281 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5341 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5401 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5461 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5521 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5581 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5641 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5701 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5761 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5821 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     5881 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     5941 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     6001 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     6061 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     6121 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6181 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6241 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6301 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6361 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6421 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6481 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6541 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6601 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6661 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6721 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6781 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6841 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6901 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6961 tgtggataac cgtattaccg ccatgcat
//
