LOCUS       sequence-347307-        6753 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of PB1-MontiRed-LOV2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6753)
  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 6753)
  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-LOV2
FEATURES             Location/Qualifiers
     source          1..6753
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        380..655
                     /label=chicken beta-actin promoter
     intron          656..1664
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1588..1610
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1734..1743
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2424..2426,2427..2429,2430..3089)
                     /codon_start=1
                     /product="humanized monomeric Keima-Red fluorescent 
                     protein"
                     /label=hmKeima-Red
                     /note="mammalian codon-optimized"
                     /translation="MVGVIAKQMTYKVYMSGTVNGHYFEVEGDGKGKPYEGEQTVRLTV
                     TKGGPLPFAWDILSPQLMYGSITFTKYPEDIPDYFKQSFPEGFTWERIMNFEDGAVCTV
                     SNDSSIQGNCFIYNVKISGVNFPPNGPVMQKKTQGWEPSTERLFARDGMLIGYNYMALK
                     LEGGGHYLCEFKTTYKAKKPVRMPGFHFIDRKLDVTSHNRDYTSVEQCEIAIARHSLLG
                     "
     CDS             3135..3563
                     /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
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNLFHLQPMRDQKGDVQ
                     YFIGVQLDGTEHVRDAAEREGVMLIKKTAENIDEAAKEL"
     primer_bind     complement(3612..3632)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3612..3630)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3651..3667)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3769..3788)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3834..3889
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3888..3907)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4250..4266)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4274..4290
                     /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(4298..4304,4305..4322,4323..4328))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4343..4364
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4423..4618
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4469..4604
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4624..4758
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4661..4680)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4715..4734
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4826..4843)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4997..5585)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5077..5096)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5756..6547,6548..6616))
                     /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     6379..6398
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6617..6721)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(6751..6753,1..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 gactatttac ggtaaactgc ccacttggca gtacatcaag
      241 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      301 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      361 tcatcgctat taccatgggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
      421 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg
      481 ggggcggggg gggggggggc gcgcgccagg cggggcgggg cgggggcgag ggcggggcgg
      541 ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt
      601 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt
      661 cgctgcgttg ccttcgcccc gtgccccgct ccgcgccgcc tcgcgccgcc cgccccggct
      721 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
      781 taattagcgc ttggtttaat gacggctcgt ttcttttctg tggctgcgtg aaagccttaa
      841 agggctccgg gagggccctt tgtgcggggg ggagcggctc ggggggtgcg tgcgtgtgtg
      901 tgtgcgtggg gagcgccgcg tgcggcccgc gctgcccggc ggctgtgagc gctgcgggcg
      961 cggcgcgggg ctttgtgcgc tccgcgtgtg cgcgagggga gcgcggccgg gggcggtgcc
     1021 ccgcggtgcg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg
     1081 ggtgagcagg gggtgtgggc gcggcggtcg ggctgtaacc cccccctgca cccccctccc
     1141 cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtgcggggcg tggcgcgggg
     1201 ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct
     1261 cgggccgggg agggctcggg ggaggggcgc ggcggccccg gagcgccggc ggctgtcgag
     1321 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
     1381 ctttgtccca aatctggcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg
     1441 gcgcgggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg
     1501 tcgccgcgcc gccgtcccct tctccatctc cagcctcggg gctgccgcag ggggacggct
     1561 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
     1621 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
     1681 tggttgttgt gctgtctcat cattttggca aagaattcat agtagctagc attgccacca
     1741 tggcgtcgct caccgtgaag gcctaccttc tgggcaagga ggacgcggcg cgcgagattc
     1801 gccgcttcag cttctgttgc agccccgagc ctgaggcgga agccgaggct gcggcgggtc
     1861 cgggaccctg cgagcggctg ctgagccggg tggccgccct gttccccgcg ctgcggcctg
     1921 gcggcttcca ggcgcactac cgcgatgagg acggggactt ggttgccttt tccagtgacg
     1981 aggaattgac aatggccatg tcctacgtga aggatgacat cttccgaatc tacattaaag
     2041 agaaaaccgg ttctgggagt ggcggtagtg gtgctggagg cagcgcaggc tctggagcag
     2101 gcggcagtgc cggaagtggc gctggagggt ctgccggatc tggagccggt ggttctgccg
     2161 gctccggcgc tggtgggagc gcttccggaa gtagtcccgg ttctgggagt ggcggtagtg
     2221 gtgctggagg cagcgcaggc tctggagcag gcggcagtgc cggaagtggc gctggagggt
     2281 ctgccggatc tggagccggt ggttctgccg gctccggcgc tggtgggagc gcttccggaa
     2341 gtagtcccgg taattccgct gacggcggcg gaggatcggg tggtagtggt ggttcaggag
     2401 gaggatcgac ccaaggagga tccatggtgg gcgtgatcgc caagcagatg acctacaagg
     2461 tgtacatgag cggcaccgtg aacggccact acttcgaggt ggagggcgac ggcaagggca
     2521 agccctacga gggcgagcag accgtgaggc tgaccgtgac caagggtggc cccctgccct
     2581 tcgcctggga catcctgagc ccccagctca tgtacggcag catcaccttc accaagtacc
     2641 ccgaggacat ccccgactac ttcaaacaga gcttccccga gggcttcacc tgggagcgca
     2701 tcatgaactt cgaggacggc gccgtgtgca ccgtgagcaa cgacagcagc atccagggca
     2761 actgcttcat ctacaacgtg aagatcagcg gcgtgaactt cccccccaac ggccccgtga
     2821 tgcagaagaa gacccagggc tgggagccca gcaccgagcg cctgttcgcc cgcgacggaa
     2881 tgctgatcgg ctacaactac atggccctga agctggaggg cggcggccac tacctgtgcg
     2941 agttcaagac cacctacaag gccaagaagc ccgtgaggat gcccggcttc cacttcatcg
     3001 accgcaagct ggacgtgacc agccacaacc gcgactacac cagcgtggag cagtgcgaga
     3061 tcgccatcgc ccgccacagc ctgctgggcg gtaccggaac tgcagcagag aattcgggaa
     3121 actcgagaac aaagcttgct actacacttg aacgtattga gaagaacttt gtcattactg
     3181 acccaagatt gccagataat cccattatat tcgcgtccga tagtttcttg cagttgacag
     3241 aatatagccg tgaagaaatt ttgggaagaa actgcaggtt tctacaaggt cctgaaactg
     3301 atcgcgcgac agtgagaaaa attagagatg ccatagataa ccaaacagag gtcactgttc
     3361 agctgattaa ttatacaaag agtggtaaaa agttctggaa cctctttcac ttgcagccta
     3421 tgcgagatca gaagggagat gtccagtact ttattggggt tcagttggat ggaactgagc
     3481 atgtccgaga tgctgccgag agagagggag tcatgctgat taagaaaact gcagaaaata
     3541 ttgatgaggc ggcaaaagaa ctttaacccg ggtaagcggc cgccaccgcg gtggagctcc
     3601 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     3661 tttcctgtgt gaaattgtta tccgctaatt cactcctcag gtgcaggctg cctatcagaa
     3721 ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc
     3781 tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag
     3841 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac
     3901 atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac
     3961 atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat
     4021 gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat
     4081 tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca
     4141 tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct
     4201 tctcttatgg agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt
     4261 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4321 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4381 tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc
     4441 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4501 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4561 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     4621 gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4681 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4741 tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4801 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4861 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4921 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4981 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5041 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5101 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5161 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     5221 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5281 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5341 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5401 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     5461 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5521 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5581 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5641 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5701 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5761 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5821 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5881 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5941 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6001 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6061 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6121 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6181 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6241 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6301 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6361 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6421 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6481 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6541 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6601 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6661 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6721 gcacatttcc ccgaaaagtg ccacctggtc gac
//
