LOCUS       sequence-199199-        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        62..365
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        366..569
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     520..540
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             608..1117
                     /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      1130..1139
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1136..1138,1139..1141,1142..1852)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1181..1202)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1442..1461)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1789..1810
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3795..3916
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3832..3851)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3886..3905
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3923..4378)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4060..4081)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4272..4291
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4405..4509
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4511..4868
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4511..4702
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4534..4554)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4719..4854
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4903..5697
                     /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             4903..5697
                     /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(4957..4976)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5567..5586
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5885..5904)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5929..5976
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6305..6893
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6794..6813
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
       61 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      121 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      181 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      241 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      301 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      361 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      421 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      481 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      541 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
      601 cgccaccatg aatggagcta taggaggtga ccttttgctc aattttcctg acatgtcggt
      661 cctagagcgc caaagggctc acctcaagta cctcaatccc acctttgatt ctcctctcgc
      721 cggcttcttt gccgattctt caatgattac cggcggcgag atggacagct atctttcgac
      781 tgccggtttg aatcttccga tgatgtacgg tgagacgacg gtggaaggtg attcaagact
      841 ctcaatttcg ccggaaacga cgcttgggac tggaaatttc aaggcagcga agtttgatac
      901 agagactaag gattgtaatg aggcggcgaa gaagatgacg atgaacagag atgacctagt
      961 agaagaagga gaagaagaga agtcgaaaat aacagagcaa aacaatggga gcacaaaaag
     1021 catcaagaag atgaaacaca aagccaagaa agaagagaac aatttctcta atgattcatc
     1081 taaagtgacg aaggaattgg agaaaacgga ttatattcat ggaccggtcg ccaccatggt
     1141 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1201 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1261 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1321 gaccaccttc ggctacggcc tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
     1381 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1441 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1501 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1561 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     1621 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     1681 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1741 gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1801 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtccggact
     1861 cagatctcga gctcaagctt cgaattctgc agtcgacatg gcgactgcga cccccgtgcc
     1921 gccgcggatg ggcagccgcg ctggcggccc caccacgccg ctgagcccca cgcgcctgtc
     1981 gcggctccag gagaaggagg agctgcgcga gctcaatgac cggctggcgg tgtacatcga
     2041 caaggtgcgc agcctggaga cggagaacag cgcgctgcag ctgcaggtga cggagcgcga
     2101 ggaggtgcgc ggccgtgagc tcaccggcct caaggcgctc tacgagaccg agctggccga
     2161 cgcgcgacgc gcgctcgacg acacggcccg cgagcgcgcc aagctgcaga tcgagctggg
     2221 caagtgcaag gcggaacacg accagctgct cctcaactat gctaagaagg aatctgatct
     2281 taatggcgcc cagatcaagc ttcgagaata tgaagcagca ctgaattcga aagatgcagc
     2341 tcttgctact gcacttggtg acaaaaaaag tttagaggga gatttggagg atctgaagga
     2401 tcagattgcc cagttggaag cctccttagc tgcagccaaa aaacagttag cagatgaaac
     2461 tttacttaaa gtagatttgg agaatcgttg tcagagcctt actgaggact tggagtttcg
     2521 caaaagcatg tatgaagagg agattaacga gaccagaagg aagcatgaaa cgcgcttggt
     2581 agaggtggat tctgggcgtc aaattgagta tgagtacaag ctggcgcaag cccttcatga
     2641 gatgagagag caacatgatg cccaagtgag gctgtataag gaggagctgg agcagactta
     2701 ccatgccaaa cttgagaatg ccagactgtc atcagagatg aatacttcta ctgtcaacag
     2761 tgccagggaa gaactgatgg aaagccgcat gagaattgag agcctttcat cccagctttc
     2821 taatctacag aaagagtcta gagcatgttt ggaaaggatt caagaattag aggacttgct
     2881 tgctaaagaa aaagacaact ctcgtcgcat gctgacagac aaagagagag agatggcgga
     2941 aataagggat caaatgcagc aacagctgaa tgactatgaa cagcttcttg atgtaaagtt
     3001 agccctggac atggaaatca gtgcttacag gaaactctta gaaggcgaag aagagaggtt
     3061 gaagctgtct ccaagccctt cttcccgtgt gacagtatcc cgagcatcct caagtcgtag
     3121 tgtacgtaca actagaggaa agcggaagag ggttgatgtg gaagaatcag aggcgagtag
     3181 tagtgttagc atctctcatt ccgcctcagc cactggaaat gtttgcatcg aagaaattga
     3241 tgttgatggg aaatttatcc gcttgaagaa cacttctgaa caggatcaac caatgggagg
     3301 ctgggagatg atcagaaaaa ttggagacac atcagtcagt tataaatata cctcaagata
     3361 tgtgctgaag gcaggccaga ctgttacaat ttgggctgca aacgctggtg tcacagccag
     3421 ccccccaact gacctcatct ggaagaacca gaactcgtgg ggcactggcg aagatgtgaa
     3481 ggttatattg aaaaattctc agggagagga ggttgctcaa agaagtacag tctttaaaac
     3541 aaccatacct gaagaagagg aggaggagga agaagcagct ggagtggttg ttgaggaaga
     3601 acttttccac cagcagggaa ccccaagagc atccaataga agctgtgcaa ttatgtaagg
     3661 gcccgggatc caccggatct agataactga tcataatcag ccataccaca tttgtagagg
     3721 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     3781 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3841 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3901 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     3961 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     4021 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     4081 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     4141 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     4201 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     4261 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     4321 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     4381 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4441 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4501 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     4561 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4621 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4681 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4741 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4801 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4861 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4921 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4981 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5041 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     5101 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5161 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5221 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5281 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5341 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5401 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     5461 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5521 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5581 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5641 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5701 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5761 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5821 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     5881 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     5941 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     6001 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     6061 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     6121 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     6181 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6241 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6301 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6361 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6421 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6481 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6541 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6601 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6661 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6721 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6781 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6841 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6901 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6961 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgca
//
