LOCUS       sequence-99714-e        6838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LINuS-flex: modular LINuS construct enabling introduction of any 
            protein encoding sequence, NES and NLS. The sequence of interest 
            replaces the LexA DBD-MBP-VP64 fragment..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6838)
  AUTHORS   Niopek D, Benzinger D, Roensch J, Draebing T, Wehler P, Eils R, Di 
            Ventura B
  TITLE     Engineering light-inducible nuclear localization signals for precise
            spatiotemporal control of protein dynamics in living cells.
  JOURNAL   Nat Commun. 2014 Jul 14;5:4404. doi: 10.1038/ncomms5404.
  PUBMED    25019686
REFERENCE   2  (bases 1 to 6838)
  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"; journalName: "Nat 
            Commun."; date: "2014-07-14"; volume: "5:4404. doi"; pages: " 
            10.1038/ncomms5404"
COMMENT     Sequence Label: pDN80
FEATURES             Location/Qualifiers
     source          1..6838
                     /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"
     misc_feature    591..671
                     /label=MCS
                     /note="multiple cloning site"
     CDS             679..930
                     /codon_start=1
                     /gene="lexA (truncated)"
                     /product="DNA-binding domain of E. coli LexA"
                     /label=LexA BD
                     /note="binds the 16-bp palindromic sequence 
                     5'-CTGTATATATATACAG-3'"
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAA"
     CDS             join(958..1032,1033..2130)
                     /codon_start=1
                     /gene="malE"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This gene encodes a signal sequence that can direct 
                     MBP to the bacterial periplasm."
                     /translation="KIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYNGL
                     AEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEI
                     TPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKE
                     LKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVD
                     LIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSK
                     PFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDP
                     RIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1987..1996
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2104..2127
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2176..2286
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     CDS             2335..3039
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2477..2495)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2745..2764
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3626..3747
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3663..3682)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3717..3736
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3754..4209)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3891..3912)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4103..4122
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4236..4340
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4342..4699
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4342..4533
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4365..4385)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4550..4685
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4734..5528
                     /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             4734..5528
                     /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(4788..4807)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5398..5417
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5716..5735)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5760..5807
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6136..6724
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6625..6644
                     /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 gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
      661 gatccaccgg tcgccaccat gaaagcgtta acggccaggc aacaagaggt gtttgatctc
      721 atccgtgatc acatcagcca gacaggtatg ccgccgacgc gtgcggaaat cgcgcagcgt
      781 ttggggttcc gttccccaaa cgcggctgaa gaacatctga aggcgctggc acgcaaaggc
      841 gttattgaaa ttgtttccgg cgcatcacgc gggattcgtc tgttgcagga agaggaagaa
      901 gggttgccgc tggtaggtcg tgtggctgcc ggtgaaccag gtggtagcgg aaccagcaaa
      961 ataaaaacag gtgcacgcat cctcgcatta tccgcattaa cgacgatgat gttttccgcc
     1021 tcggctctcg ccaaaatcga agaaggtaaa ctggtaatct ggattaacgg cgataaaggc
     1081 tataacggcc tcgctgaagt cggtaagaaa ttcgagaaag ataccggaat taaagtcacc
     1141 gttgagcatc cggataaact ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc
     1201 cctgacatta tcttctgggc acacgaccgc tttggtggct acgctcaatc tggcctgttg
     1261 gctgaaatca ccccggacaa agcgttccag gacaagctgt atccgtttac ctgggatgcc
     1321 gtacgttaca acggcaagct gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt
     1381 tataacaaag atctgctgcc gaacccgcca aaaacctggg aagagatccc ggcgctggat
     1441 aaagaactga aagcgaaagg taagagcgcg ctgatgttca acctgcaaga accgtacttc
     1501 acctggccgc tgattgctgc tgacgggggt tatgcgttca agtatgaaaa cggcaagtac
     1561 gacattaaag acgtgggcgt ggataacgct ggcgcgaaag cgggtctgac cttcctggtt
     1621 gacctgatta aaaacaaaca catgaatgca gacaccgatt actccatcgc agaagctgcc
     1681 tttaataaag gcgaaacagc gatgaccatc aacggcccgt gggcatggtc caacatcgac
     1741 accagcaaag tgaattatgg tgtaacggta ctgccgacct tcaagggtca accatccaaa
     1801 ccgttcgttg gcgtgctgag cgcaggtatt aacgccgcca gtccgaacaa agagctggcg
     1861 aaagagttcc tcgaaaacta tctgctgact gatgaaggtc tggaagcggt taataaagac
     1921 aaaccgctgg gtgccgtagc gctgaagtct tacgaggaag agttggcgaa agatccacgt
     1981 attgccgcca ccatggaaaa cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg
     2041 tccgctttct ggtatgccgt gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact
     2101 gtcgatgaag ccctgaaaga cgcgcagact cgtatcacca agggcggcag cggtaccgaa
     2161 agcggcgggc cggccgacgc ccttgacgat tttgacttag acatgctccc agccgatgcc
     2221 cttgacgact ttgaccttga tatgctgcct gctgacgctc ttgacgattt tgaccttgac
     2281 atgctccccg ggactagtgg agacgccggc ggcagcggcg gcagcgtcgt ctcagtgagc
     2341 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
     2401 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     2461 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
     2521 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
     2581 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     2641 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
     2701 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
     2761 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
     2821 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
     2881 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
     2941 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
     3001 ggccgccact ccaccggcgg catggacgag ctgtacaagg gtggatctgg aggttcaggt
     3061 ggaagtttgg ctactacact tgaacgtatt gagaagaact ttgtcattac tgacccaaga
     3121 ttgccagata atcccattat attcgcgtcc gatagtttct tgcagttgac agaatatagc
     3181 cgtgaagaaa ttttgggaag aaactgcagg tttctacaag gtcctgaaac tgatcgcgcg
     3241 acagtgagaa aaattagaga tgccatagat aaccaaacag aggtcactgt tcagctgatt
     3301 aattatacaa agagtggtaa aaagttctgg aacctctttc acttgcagcc tatgcgagat
     3361 cagaagggag atgtccagta ctttattggg gttcagttgg atggaactga gcatgtccga
     3421 gatgctgccg agagagaggg agtcatgctg attaagaaaa ctgcagaaaa tattggagac
     3481 caatagtaga aattaaaggt ctcagcggcc gcgactctag atcataatca gccataccac
     3541 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     3601 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     3661 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3721 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
     3781 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     3841 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     3901 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     3961 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     4021 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     4081 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     4141 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     4201 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4261 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4321 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     4381 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4441 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4501 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4561 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4621 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     4681 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     4741 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4801 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4861 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     4921 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4981 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5041 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5101 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5161 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5221 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     5281 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5341 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5401 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5461 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5521 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5581 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5641 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5701 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     5761 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     5821 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     5881 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     5941 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     6001 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6061 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6121 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6181 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6241 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     6301 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6361 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6421 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6481 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6541 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6601 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6661 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6721 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6781 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
