LOCUS       sequence-228919-        6652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mCherry fused to LINuS, a light-inducible nuclear localization 
            signal (biNLS2/PKIt NES).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6652)
  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 6652)
  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: pDN34
FEATURES             Location/Qualifiers
     source          1..6652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             934..1638
                     /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(1076..1094)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1344..1363
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2243..2260)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2249..2473
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2519..2947
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2606..2625)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2816..2837
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2956..2976)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2961..3290
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3141..3276
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3203..3222
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3357..4151
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3411..3430)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4021..4040
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4325..4446
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4362..4381)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4416..4435
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4495..4511)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4495..4511)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4508..4530)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4519..4535
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4543..4549,4550..4567,4568..4573))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4588..4609
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4726..4743)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4897..5485)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4977..4996)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5656..6447,6448..6516))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6279..6298
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6517..6621)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 atgttagcct tgaaattagc aggtcttgat atcgtgagca agggcgagga ggataacatg
      961 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
     1021 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     1081 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
     1141 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
     1201 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     1261 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
     1321 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     1381 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
     1441 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
     1501 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
     1561 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     1621 atggacgagc tgtacaaggg tggatctgga ggttcaggtg gaagtttggc tactacactt
     1681 gaacgtattg agaagaactt tgtcattact gacccaagat tgccagataa tcccattata
     1741 ttcgcgtccg atagtttctt gcagttgaca gaatatagcc gtgaagaaat tttgggaaga
     1801 aactgcaggt ttctacaagg tcctgaaact gatcgcgcga cagtgagaaa aattagagat
     1861 gccatagata accaaacaga ggtcactgtt cagctgatta attatacaaa gagtggtaaa
     1921 aagttctgga acctctttca cttgcagcct atgcgagatc agaagggaga tgtccagtac
     1981 tttattgggg ttcagttgga tggaactgag catgtccgag atgctgccga gagagaggga
     2041 gtcatgctga ttaagaaaac tgcagaaaat attgacgagg ccgcaaagag actgcccgac
     2101 gccaacctgg cagccgcagc caagaagaaa aagctggact agctcgagcg gccgccactg
     2161 tgctggatat ctgcagaatt ccaccacact ggactagtgg atccgagctc ggtaccaagc
     2221 ttaagtttaa accgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2281 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2341 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2401 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     2461 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2521 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2581 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2641 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2701 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2761 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2821 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2881 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2941 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3001 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3061 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3121 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3181 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3241 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3301 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3361 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3421 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3481 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3541 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3601 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3661 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3721 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3781 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3841 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3901 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3961 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4021 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4081 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4141 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4201 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4261 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4321 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4381 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4441 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4501 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4561 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4621 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4681 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4741 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4801 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4861 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4921 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4981 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5041 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5101 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5161 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5221 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5281 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5341 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5401 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5461 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5521 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5581 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5641 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5701 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5761 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5821 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5881 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5941 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6001 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6061 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6121 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6181 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6241 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6301 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6361 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6421 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6481 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6541 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6601 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
