LOCUS       sequence-351718-        6761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of PB1-MontiRed-LOV2dark.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6761)
  AUTHORS   Yoshikawa M, Yoshii T, Ikuta M, Tsukiji S
  TITLE     Synthetic Protein Condensates That Inducibly Recruit and Release 
            Protein Activity in Living Cells.
  JOURNAL   J Am Chem Soc. 2021 May 5;143(17):6434-6446. doi: 
            10.1021/jacs.0c12375. Epub 2021 Apr 23.
  PUBMED    33890764
REFERENCE   2  (bases 1 to 6761)
  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: 
            "10.1021/jacs.0c12375"; journalName: "J Am Chem Soc"; date: 
            "2021-05-5- 5"; volume: "143"; issue: "17"; pages: "6434-6446"
COMMENT     Sequence Label: pCAGGS-PB1-MontiRed-LOV2dark
FEATURES             Location/Qualifiers
     source          1..6761
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          655..1671
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1595..1617
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1679..1698
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1741..1750
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2431..2433,2434..2436,2437..3096)
                     /codon_start=1
                     /product="humanized monomeric Keima-Red fluorescent 
                     protein"
                     /label=hmKeima-Red
                     /note="mammalian codon-optimized"
                     /translation="MVGVIAKQMTYKVYMSGTVNGHYFEVEGDGKGKPYEGEQTVRLTV
                     TKGGPLPFAWDILSPQLMYGSITFTKYPEDIPDYFKQSFPEGFTWERIMNFEDGAVCTV
                     SNDSSIQGNCFIYNVKISGVNFPPNGPVMQKKTQGWEPSTERLFARDGMLIGYNYMALK
                     LEGGGHYLCEFKTTYKAKKPVRMPGFHFIDRKLDVTSHNRDYTSVEQCEIAIARHSLLG
                     "
     CDS             3142..3570
                     /codon_start=1
                     /gene="NPH1 from Avena sativa"
                     /product="light, oxygen, or voltage domain from phototropin
                     1 of Avena sativa (Herrou and Crosson, 2011)"
                     /label=LOV2
                     /note="photoswitch that undergoes a light-inducible 
                     conformational change"
                     /translation="LATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NARFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNLFHLQPMRDQKGDVQ
                     YFIGVQKDGTEHVRDAAEREAVMEIKKTAEEIDEAAKEL"
     primer_bind     complement(3619..3639)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3619..3637)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3658..3674)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3776..3795)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3841..3896
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3895..3914)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4257..4273)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4281..4297
                     /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(4305..4311,4312..4329,4330..4335))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4350..4371
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4430..4625
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4476..4611
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4631..4765
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4668..4687)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4722..4741
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4833..4850)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5004..5592)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5084..5103)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5763..6554,6555..6623))
                     /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     6386..6405
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6624..6728)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(6759..6761,1..377)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
       61 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      121 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      181 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      241 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      301 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      361 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
      421 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
      481 gggcgggggg ggggggggsg cgcgccrggg gggggggggg gggggggggg ggggggsggg
      541 gsgrggsggr gaggkgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
      601 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
      661 gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg
      721 ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc
      781 tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt
      841 gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt
      901 gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg
      961 cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt
     1021 gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg
     1081 ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct
     1141 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg
     1201 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
     1261 cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt
     1321 cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga
     1381 cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct
     1441 agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg
     1561 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     1621 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     1681 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattcatagt agctagcatt
     1741 gccaccatgg cgtcgctcac cgtgaaggcc taccttctgg gcaaggagga cgcggcgcgc
     1801 gagattcgcc gcttcagctt ctgttgcagc cccgagcctg aggcggaagc cgaggctgcg
     1861 gcgggtccgg gaccctgcga gcggctgctg agccgggtgg ccgccctgtt ccccgcgctg
     1921 cggcctggcg gcttccaggc gcactaccgc gatgaggacg gggacttggt tgccttttcc
     1981 agtgacgagg aattgacaat ggccatgtcc tacgtgaagg atgacatctt ccgaatctac
     2041 attaaagaga aaaccggttc tgggagtggc ggtagtggtg ctggaggcag cgcaggctct
     2101 ggagcaggcg gcagtgccgg aagtggcgct ggagggtctg ccggatctgg agccggtggt
     2161 tctgccggct ccggcgctgg tgggagcgct tccggaagta gtcccggttc tgggagtggc
     2221 ggtagtggtg ctggaggcag cgcaggctct ggagcaggcg gcagtgccgg aagtggcgct
     2281 ggagggtctg ccggatctgg agccggtggt tctgccggct ccggcgctgg tgggagcgct
     2341 tccggaagta gtcccggtaa ttccgctgac ggcggcggag gatcgggtgg tagtggtggt
     2401 tcaggaggag gatcgaccca aggaggatcc atggtgggcg tgatcgccaa gcagatgacc
     2461 tacaaggtgt acatgagcgg caccgtgaac ggccactact tcgaggtgga gggcgacggc
     2521 aagggcaagc cctacgaggg cgagcagacc gtgaggctga ccgtgaccaa gggtggcccc
     2581 ctgcccttcg cctgggacat cctgagcccc cagctcatgt acggcagcat caccttcacc
     2641 aagtaccccg aggacatccc cgactacttc aaacagagct tccccgaggg cttcacctgg
     2701 gagcgcatca tgaacttcga ggacggcgcc gtgtgcaccg tgagcaacga cagcagcatc
     2761 cagggcaact gcttcatcta caacgtgaag atcagcggcg tgaacttccc ccccaacggc
     2821 cccgtgatgc agaagaagac ccagggctgg gagcccagca ccgagcgcct gttcgcccgc
     2881 gacggaatgc tgatcggcta caactacatg gccctgaagc tggagggcgg cggccactac
     2941 ctgtgcgagt tcaagaccac ctacaaggcc aagaagcccg tgaggatgcc cggcttccac
     3001 ttcatcgacc gcaagctgga cgtgaccagc cacaaccgcg actacaccag cgtggagcag
     3061 tgcgagatcg ccatcgcccg ccacagcctg ctgggcggta ccggaactgc agcagagaat
     3121 tcgggaaact cgagaacaaa gcttgctact acacttgaac gtattgagaa gaactttgtc
     3181 attactgacc caagattgcc agataatccc attatattcg cgtccgatag tttcttgcag
     3241 ttgacagaat atagccgtga agaaattttg ggaagaaacg ccaggtttct acaaggtcct
     3301 gaaactgatc gcgcgacagt gagaaaaatt agagatgcca tagataacca aacagaggtc
     3361 actgttcagc tgattaatta tacaaagagt ggtaaaaagt tctggaacct ctttcacttg
     3421 cagcctatgc gagatcagaa gggagatgtc cagtacttta ttggggttca gaaggatgga
     3481 actgagcatg tccgagatgc tgccgagaga gaggccgtca tggagattaa gaaaactgca
     3541 gaagagattg atgaggcggc aaaagaactt taacccgggt aagcggccgc caccgcggtg
     3601 gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     3661 atagctgttt cctgtgtgaa attgttatcc gctaattcac tcctcaggtg caggctgcct
     3721 atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt
     3781 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
     3841 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     3901 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
     3961 tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc
     4021 agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag
     4081 gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt
     4141 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
     4201 tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca
     4261 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4321 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4381 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt
     4441 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4501 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4561 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4621 gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4681 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4741 caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga
     4801 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4861 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4921 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4981 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5041 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5101 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5161 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5221 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5281 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5341 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5401 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5461 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5521 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5581 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5641 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5701 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5761 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5821 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5881 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5941 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6001 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6061 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6121 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6181 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6241 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6301 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6361 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6421 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6481 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6541 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6601 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6661 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6721 gttccgcgca catttccccg aaaagtgcca cctgggtcga c
//
