LOCUS       sequence-351712-        6776 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of PB1-AzamiGreen(Y63L)-LOV2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6776)
  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 6776)
  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-AG(Y63L)-LOV2
FEATURES             Location/Qualifiers
     source          1..6776
                     /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             3154..3582
                     /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(3634..3654)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3634..3652)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3673..3689)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3791..3810)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3856..3911
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3910..3929)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4272..4288)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4296..4312
                     /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(4320..4326,4327..4344,4345..4350))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4365..4386
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4445..4640
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4491..4626
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4646..4780
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4683..4702)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4737..4756
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4848..4865)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5019..5607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5099..5118)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5778..6569,6570..6638))
                     /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     6401..6420
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6639..6743)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(6774..6776,1..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 atggtgagcg tgatcaagcc cgagatgaag
     2461 atcaagctgt gcatgagggg caccgtgaac ggccacaact tcgtgatcga gggcgagggc
     2521 aagggcaacc cctacgaggg cacccagatc ctggacctga acgtgaccga gggcgccccc
     2581 ctgcccttcg cctacgacat cctgaccacc gtgttccagc tgggcaacag ggccttcacc
     2641 aagtaccccg ccgacatcca ggactacttc aagcagacct tccccgaggg ctaccactgg
     2701 gagaggagca tgacctacga ggaccagggc atctgcaccg ccaccagcaa catcagcatg
     2761 aggggcgact gcttcttcta cgacatcagg ttcgacggcg tgaacttccc ccccaacggc
     2821 cccgtgatgc agaagaagac cctgaagtgg gagcccagca ccgagaagat gtacgtgagg
     2881 gacggcgtgc tgaagggcga cgtgaacatg gccctgctgc tggagggcgg cggccactac
     2941 aggtgcgact tcaagaccac ctacaaggcc aagaaggacg tgaggctgcc cgactaccac
     3001 ttcgtggacc acaggatcga gatcctgaag cacgacaagg actacaacaa ggtgaagctg
     3061 tacgagaacg ccgtggccag gtacagcatg ctgcccagcc aggccaaggg taccggaact
     3121 gcagcagaga attcgggaaa ctcgagaaca aagcttgcta ctacacttga acgtattgag
     3181 aagaactttg tcattactga cccaagattg ccagataatc ccattatatt cgcgtccgat
     3241 agtttcttgc agttgacaga atatagccgt gaagaaattt tgggaagaaa ctgcaggttt
     3301 ctacaaggtc ctgaaactga tcgcgcgaca gtgagaaaaa ttagagatgc catagataac
     3361 caaacagagg tcactgttca gctgattaat tatacaaaga gtggtaaaaa gttctggaac
     3421 ctctttcact tgcagcctat gcgagatcag aagggagatg tccagtactt tattggggtt
     3481 cagttggatg gaactgagca tgtccgagat gctgccgaga gagagggagt catgctgatt
     3541 aagaaaactg cagaaaatat tgatgaggcg gcaaaagaac tttaacccgg gatactcgcg
     3601 gccgccaccg cggtggagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     3661 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctaa ttcactcctc
     3721 aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat
     3781 accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
     3841 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     3901 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
     3961 atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc
     4021 tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa
     4081 aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa
     4141 catccctaaa attttcctta catgttttac tagccagatt tttcctcctc tcctgactac
     4201 tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag cccaagcttg
     4261 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4321 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4381 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg
     4441 atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4501 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4561 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4621 gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata
     4681 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4741 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc
     4801 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4861 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4921 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4981 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5041 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5101 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5161 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5221 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5281 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5341 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5401 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5461 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5521 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5581 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5641 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5701 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5761 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5821 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5881 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5941 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6001 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6061 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6121 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6181 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6241 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6301 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6361 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6421 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6481 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6541 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6601 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6661 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6721 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgg gtcgac
//
