LOCUS       sequence-203431-        6502 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Soluble BLInCR transcription activator (transactivation domain of 
            viral VP16 protein) that is recruited to 'localizer' sites upon blue
            light illumination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6502)
  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 6502)
  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: pPHR-mCherry-NLS-VP16
FEATURES             Location/Qualifiers
     source          1..6502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        54..158
                     /gene="bla"
                     /label=AmpR promoter
     promoter        160..517
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        160..351
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(183..203)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      368..503
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             552..1346
                     /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             552..1346
                     /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(606..625)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1216..1235
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1534..1553)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1578..1625
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1954..2542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2443..2462
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2717..3020
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3021..3224
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3175..3195
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3281..4774
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWHAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     regulatory      4793..4802
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4799..5506
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4944..4962)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5212..5231
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             5534..5554
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5946..6067
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5983..6002)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6037..6056
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(join(6074..6502,1..27))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6211..6232)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6423..6442
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
       61 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
      121 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
      181 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
      241 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
      301 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
      361 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
      421 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
      481 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
      541 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
      601 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
      661 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
      721 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
      781 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
      841 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
      901 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
      961 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     1021 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     1081 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     1141 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     1201 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     1261 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     1321 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     1381 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     1441 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     1501 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     1561 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     1621 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     1681 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     1741 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     1801 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1861 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1921 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1981 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2041 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2101 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2161 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2221 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2281 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2341 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2401 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2461 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2521 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2581 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     2641 tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     2701 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     2761 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     2821 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     2881 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     2941 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     3001 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     3061 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     3121 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     3181 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     3241 agatccgcta gcgctaccgg actcagatct cgaggccacc atgaagatgg acaaaaagac
     3301 tatagtttgg tttagaagag acctaaggat tgaggataat cctgcattag cagcagctgc
     3361 tcacgaagga tctgtttttc ctgtcttcat ttggtgtcct gaagaagaag gacagtttta
     3421 tcctggaaga gcttcaagat ggtggatgaa acaatcactt gctcacttat ctcaatcctt
     3481 gaaggctctt ggatctgacc tcactttaat caaaacccac aacacgattt cagcgatctt
     3541 ggattgtatc cgcgttaccg gtgctacaaa agtcgtcttt aaccacctct atgatcctgt
     3601 ttcgttagtt cgggaccata ccgtaaagga gaagctggtg gaacgtggga tctctgtgca
     3661 aagctacaat ggagatctat tgtatgaacc gtgggagata tactgcgaaa agggcaaacc
     3721 ttttacgagt ttcaattctt actggaagaa atgcttagat atgtcgattg aatccgttat
     3781 gcttcctcct ccttggcggt tgatgccaat aactgcagcg gctgaagcga tttgggcgtg
     3841 ttcgattgaa gaactagggc tggagaatga ggccgagaaa ccgagcaatg cgttgttaac
     3901 tagagcttgg tctccaggat ggagcaatgc tgataagtta ctaaatgagt tcatcgagaa
     3961 gcagttgata gattatgcaa agaacagcaa gaaagttgtt gggaattcta cttcactact
     4021 ttctccgtat ctccatttcg gggaaataag cgtcagacac gttttccagt gtgcccggat
     4081 gaaacaaatt atatgggcaa gagataagaa cagtgaagga gaagaaagtg cagatctttt
     4141 tcttagggga atcggtttaa gagagtattc tcggtatata tgtttcaact tcccgtttac
     4201 tcacgagcaa tcgttgttga gtcatcttcg gtttttccct tgggatgctg atgttgataa
     4261 gttcaaggcc tggagacaag gcaggaccgg ttatccgttg gtggatgccg gaatgagaga
     4321 gctttgggct accggatgga tgcataacag aataagagtg attgtttcaa gctttgctgt
     4381 gaagtttctt ctccttccat ggaaatgggg aatgaagtat ttctgggata cacttttgga
     4441 tgctgatttg gaatgtgaca tccttggctg gcagtatatc tctgggagta tccccgatgg
     4501 ccacgagctt gatcgcttgg acaatcccgc gttacaaggc gccaaatatg acccagaagg
     4561 tgagtacata aggcaatggc ttcccgagct tgcgagattg ccaactgaat ggatccatca
     4621 tccatggcac gctcctttaa ccgtactcaa agcttctggt gtggaactcg gaacaaacta
     4681 tgcgaaaccc attgtagaca tcgacacagc tcgtgagcta ctagctaaag ctatttcaag
     4741 aacccgtgaa gcacagatca tgatcggagc agcagcccgg gatccaccgg tcgccaccat
     4801 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     4861 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     4921 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     4981 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     5041 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     5101 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     5161 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     5221 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     5281 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     5341 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     5401 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     5461 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagggta gggggcgtcg
     5521 actgggtacc cctcctaaga agaagaggaa ggttgccccc accatggata aaaagaagag
     5581 aaaggccccc ccgaccgatg tcagcctggg ggacgagctc cacttagacg gcgaggacgt
     5641 ggcgatggcg catgccgacg cgctagacga tttcgatctg gacatgttgg gggacgggga
     5701 ttccccgggt ccgggattta ccccccacga ctccgccccc tacggcgctc tggatatggc
     5761 cgacttcgag tttgagcaga tgtttaccga tgcccctcta gacgggggga tccaccggat
     5821 ctaggcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     5881 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     5941 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     6001 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     6061 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     6121 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     6181 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     6241 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     6301 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     6361 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     6421 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     6481 cgcgtaacca ccacacccgc cg
//
