LOCUS       sequence-271001-        6107 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of iLID fused to mCherry and intracellular 
            signaling domain of TrkA (aa 450-799) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6107)
  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 6107)
  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-mCherry-iTrkA
FEATURES             Location/Qualifiers
     source          1..6107
                     /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             631..1068
                     /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      1105..1114
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1111..1818
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1256..1274)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1524..1543
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2895..3016
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2932..2951)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2986..3005
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3023..3478)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3160..3181)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3372..3391
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3505..3609
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3611..3968
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3611..3802
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3634..3654)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3819..3954
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4003..4797
                     /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             4003..4797
                     /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(4057..4076)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4667..4686
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4985..5004)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5029..5076
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5405..5993
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5894..5913
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 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 atctcgagat gggggagttt ctggcaacca cactggaacg gatcgagaaa
      661 aatttcgtga ttactgatcc gagactgcct gacaacccaa tcatttttgc gagcgattcc
      721 ttcctgcagc tgacagaata ttctcgggaa gagatcctgg ggcgcaattg ccgttttctg
      781 cagggacccg agacagaccg tgccactgtt cggaaaatca gagatgctat tgacaaccag
      841 actgaagtga ccgttcagct gatcaattat accaagagcg gcaagaagtt ctggaacgtg
      901 ttccacctgc agccgatgcg cgattataag ggcgacgtcc agtacttcat tggcgtgcag
      961 ctggatggca ccgaacgtct tcatggcgcc gctgagcgtg aggcggtctg cctgatcaaa
     1021 aagacagcct ttcagattgc tgaggcagcg aacgacgaaa attactttgg aagcgggagt
     1081 gggagccccc gggatccacc ggtcgccacc atggtgagca agggcgagga ggataacatg
     1141 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
     1201 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     1261 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
     1321 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
     1381 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     1441 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
     1501 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     1561 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
     1621 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
     1681 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
     1741 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     1801 atggacgagc tgtacaaggg ttcttctaaa tttgggatca accgccctgc tgtgctggcg
     1861 ccagaggatg ggctggccat gtccctacac ttcatgacac tgggtggcag ttctctttcc
     1921 cctactgagg gcaaaggctc cggactccag ggccacatca tggagaaccc acagtacttc
     1981 agtgatacct gtgtccacca tatcaagcgc caggacatca ttctcaagtg ggagctaggg
     2041 gagggagcct ttggaaaggt ctttcttgct gagtgctaca accttctgaa tgatcaggac
     2101 aagatgctgg tggctgtcaa ggcactgaag gagacatctg agaatgctcg tcaggacttc
     2161 catcgtgagg cagagctgct caccatgcta cagcaccaac acatcgtacg cttctttgga
     2221 gtctgcacgg agggtggccc attgctcatg gtcttcgagt acatgcgcca tggggacctc
     2281 aaccgtttcc tccggtccca cggacctgat gcaaaactgc tggctggcgg cgaggatgtg
     2341 gctcctggtc ctttgggcct tgggcagctt ctggctgtgg ctagccaggt ggctgctggt
     2401 atggtgtacc tagccagcct gcactttgtg caccgggacc tggccacacg caactgtctg
     2461 gtgggtcagg gactagtggt caagattgga gacttcggca tgagcaggga catctacagc
     2521 acagactact accgtgtggg aggtcggacc atgctgccca tccgctggat gcctccagag
     2581 agcatcctct accgcaagtt cagcaccgag agtgatgtgt ggagcttcgg ggtggtgctc
     2641 tgggagatct tcacctatgg aaagcaaccc tggtaccagc tctccaacac tgaggcgatc
     2701 gagtgcatca cgcagggccg ggagctggag cggccgcgcg cctgccctcc tgatgtctac
     2761 gccatcatgc gcggctgctg gcagcgtgag ccgcaacagc gcctcagcat gaaggatgtg
     2821 cacgcgcggc tgcaggcctt ggcacaggcg ccaccgagtt acctggacgt tctgggttag
     2881 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2941 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3001 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     3061 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3121 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3181 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3241 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3301 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3361 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     3421 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     3481 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3541 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3601 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     3661 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3721 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3781 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3841 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3901 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3961 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4021 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4081 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4141 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     4201 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4261 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4321 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4381 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4441 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4501 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     4561 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4621 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4681 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4741 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4801 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4861 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4921 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     4981 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     5041 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     5101 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     5161 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     5221 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     5281 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5341 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5401 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5461 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5521 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5581 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5641 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5701 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5761 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5821 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5881 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5941 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6001 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6061 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
