LOCUS       sequence-271003-        6310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of iLID fused to mRuby3 and intracellular 
            signaling domain of TrkB (aa 455-822).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6310)
  AUTHORS   Hope JM, Liu A, Calvin GJ, Cui B
  TITLE     Construction of Light-Activated Neurotrophin Receptors Using the 
            Improved Light-Induced Dimerizer (iLID).
  JOURNAL   J Mol Biol. 2020 Jun 12;432(13):3739-3748. doi: 
            10.1016/j.jmb.2020.04.018. Epub 2020 Apr 23.
  PUBMED    32335036
REFERENCE   2  (bases 1 to 6310)
  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: 
            "10.1016/j.jmb.2020.04.018"; journalName: "J Mol Biol"; date: 
            "2020-06-12- 12"; volume: "432"; issue: "13"; pages: "3739-3748"
COMMENT     Sequence Label: iLID-mRuby3-iTrkB
FEATURES             Location/Qualifiers
     source          1..6310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             647..1084
                     /codon_start=1
                     /product="improved light-inducible dimer when used with 
                     SspB nano or SspB micro (Guntas et al., 2015)"
                     /label=iLID
                     /translation="LATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNVFHLQPMRDYKGDVQ
                     YFIGVQLDGTERLHGAAEREAVCLIKKTAFQIAEAANDENYF"
     regulatory      1121..1130
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1127..1837
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     polyA_signal    3098..3219
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3135..3154)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3189..3208
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3226..3681)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3363..3384)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3575..3594
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3708..3812
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3814..4171
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3814..4005
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3837..3857)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4022..4157
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4206..5000
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4206..5000
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4260..4279)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4870..4889
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5188..5207)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5232..5279
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5608..6196
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6097..6116
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatatg tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcatgggg gagtttctgg caaccacact
      661 ggaacggatc gagaaaaatt tcgtgattac tgatccgaga ctgcctgaca acccaatcat
      721 ttttgcgagc gattccttcc tgcagctgac agaatattct cgggaagaga tcctggggcg
      781 caattgccgt tttctgcagg gacccgagac agaccgtgcc actgttcgga aaatcagaga
      841 tgctattgac aaccagactg aagtgaccgt tcagctgatc aattatacca agagcggcaa
      901 gaagttctgg aacgtgttcc acctgcagcc gatgcgcgat tataagggcg acgtccagta
      961 cttcattggc gtgcagctgg atggcaccga acgtcttcat ggcgccgctg agcgtgaggc
     1021 ggtctgcctg atcaaaaaga cagcctttca gattgctgag gcagcgaacg acgaaaatta
     1081 ctttggaagc gggagtggga gcccccggga tccaccggtc gccaccatgg tgtctaaggg
     1141 cgaagagctg atcaaggaaa atatgcgtat gaaggtggtc atggaaggtt cggtcaacgg
     1201 ccaccaattc aaatgcacag gtgaaggaga aggcagaccg tacgagggag tgcaaaccat
     1261 gaggatcaaa gtcatcgagg gaggacccct gccatttgcc tttgacattc ttgccacgtc
     1321 gttcatgtat ggcagccgta cctttatcaa gtacccggcc gacatccctg atttctttaa
     1381 acagtccttt cctgagggtt ttacttggga aagagttacg agatacgaag atggtggagt
     1441 cgtcaccgtc acgcaggaca ccagccttga ggatggcgag ctcgtctaca acgtcaaggt
     1501 cagaggggta aactttccct ccaatggtcc cgtgatgcag aagaagacca agggttggga
     1561 gcctaataca gagatgatgt atccagcaga tggtggtctg agaggataca ctgacatcgc
     1621 actgaaagtt gatggtggtg gccatctgca ctgcaacttc gtgacaactt acaggtcaaa
     1681 aaagaccgtc gggaacatca agatgcccgg tgtccatgcc gttgatcacc gcctggaaag
     1741 gatcgaggag agtgacaatg aaacctacgt agtgcaaaga gaagtggcag ttgccaaata
     1801 cagcaacctt ggtggtggca tggacgagct gtacaagtcc ggactcagat ctggaggctc
     1861 cggaggcaag ttggcgagac attccaagtt tggcatgaaa ggcccagcct ccgttatcag
     1921 caatgatgat gactctgcca gcccactcca tcacatctcc aatgggagta acactccatc
     1981 ttcttcggaa ggtggcccag atgctgtcat tattggaatg accaagatcc ctgtcattga
     2041 aaatccccag tactttggca tcaccaacag tcagctcaag ccagacacat ttgttcagca
     2101 catcaagcga cataacattg ttctgaaaag ggagctaggc gaaggagcct ttggaaaagt
     2161 gttcctagct gaatgctata acctctgtcc tgagcaggac aagatcttgg tggcagtgaa
     2221 gaccctgaag gatgccagtg acaatgcacg caaggacttc caccgtgagg ccgagctcct
     2281 gaccaacctc cagcatgagc acatcgtcaa gttctatggc gtctgcgtgg agggcgaccc
     2341 cctcatcatg gtctttgagt acatgaagca tggggacctc aacaagttcc tcagggcaca
     2401 cggccctgat gccgtgctga tggctgaggg caacccgccc acggaactga cgcagtcgca
     2461 gatgctgcat atagcccagc agatcgccgc gggcatggtc tacctggcgt cccagcactt
     2521 cgtgcaccgc gatttggcca ccaggaactg cctggtcggg gagaacttgc tggtgaaaat
     2581 cggggacttt gggatgtccc gggacgtgta cagcactgac tactacaggg tcggtggcca
     2641 cacaatgctg cccattcgct ggatgcctcc agagagcatc atgtacagga aattcacgac
     2701 ggaaagcgac gtctggagcc tgggggtcgt gttgtgggag attttcacct atggcaaaca
     2761 gccctggtac cagctgtcaa acaatgaggt gatagagtgt atcactcagg gccgagtcct
     2821 gcagcgaccc cgcacgtgcc cccaggaggt gtatgagctg atgctggggt gctggcagcg
     2881 agagccccac atgaggaaga acatcaaggg catccatacc ctccttcaga acttggccaa
     2941 ggcatctccg gtctacctgg acatcctagg ctagaagcgg ccgcgactct agatcataat
     3001 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     3061 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     3121 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3181 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     3241 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3301 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3361 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3421 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3481 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3541 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3601 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3661 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3721 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3781 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     3841 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3901 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3961 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4021 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4081 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4141 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     4201 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4261 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4321 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4381 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4441 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4501 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4561 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4621 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4681 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     4741 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4801 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4861 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4921 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4981 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     5041 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5101 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5161 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     5221 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     5281 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     5341 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     5401 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     5461 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5521 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5581 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5641 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5701 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5761 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5821 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5881 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5941 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6001 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6061 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6121 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6181 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6241 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6301 cgccatgcat
//
