LOCUS       Exported                6824 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  mCherry fused to LINuS, a light-inducible nuclear localization 
            signal (biNLS9/PKIt NES).
ACCESSION   .
VERSION     .
KEYWORDS    pDN41
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6824)
  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 6824)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6824
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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(2249..2266)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2255..2479
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2525..2953
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2612..2631)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2822..2843
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2962..2982)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2967..3296
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3147..3282
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3209..3228
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3345..4370
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    4500..4621
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4537..4556)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4591..4610
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4670..4686)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4670..4686)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4683..4705)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4694..4710
                     /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(4718..4748)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4763..4784
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4901..4918)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5072..5657)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5152..5171)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5828..6688)
                     /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     6451..6470
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6689..6793)
                     /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 ccgcagcagc cgcagccaag aagaaaaagc tggactagct cgagcggccg
     2161 ccactgtgct ggatatctgc agaattccac cacactggac tagtggatcc gagctcggta
     2221 ccaagcttaa gtttaaaccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     2281 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2341 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2401 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     2461 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     2521 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2581 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2641 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2701 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2761 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2821 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2881 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2941 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3001 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3061 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3121 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3181 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     3241 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3301 tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgatgaaa aagcctgaac
     3361 tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga
     3421 tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat
     3481 atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc
     3541 actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa ttcagcgaga
     3601 gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa
     3661 ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg
     3721 atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta
     3781 catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga
     3841 tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg
     3901 aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga
     3961 cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc
     4021 aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga
     4081 cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata
     4141 tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg
     4201 cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc
     4261 gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg
     4321 ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag cacgtgctac
     4381 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4441 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4501 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4561 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4621 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4681 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4741 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4801 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4861 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4921 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4981 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5041 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5101 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5161 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5221 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5281 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5341 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5401 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5461 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5521 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5581 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca
     5641 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5701 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5761 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5821 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5881 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5941 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6001 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6061 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6121 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6181 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6241 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6301 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6361 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6421 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6481 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6541 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6601 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6661 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6721 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6781 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
