LOCUS       sequence-208647-        7139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of CRY2PHR (aa 1-498) fused to mCherry and 
            intracellular domain of TrkA (aa 450-799).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7139)
  AUTHORS   Duan L, Hope JM, Guo S, Ong Q, Francois A, Kaplan L, Scherrer G, Cui
            B
  TITLE     Optical Activation of TrkA Signaling.
  JOURNAL   ACS Synth Biol. 2018 Jul 20;7(7):1685-1693. doi: 
            10.1021/acssynbio.8b00126. Epub 2018 Jul 12.
  PUBMED    29975841
REFERENCE   2  (bases 1 to 7139)
  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/acssynbio.8b00126"; journalName: "ACS Synth Biol"; date: 
            "2018-07-20- 20"; volume: "7"; issue: "7"; pages: "1685-1693"
COMMENT     Sequence Label: CRY2-mCherry-iTrkA
FEATURES             Location/Qualifiers
     source          1..7139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             565..2058
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     regulatory      2077..2086
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2083..2790
                     /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(2228..2246)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2496..2515
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3867..3988
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3904..3923)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3958..3977
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3995..4450)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4132..4153)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4344..4363
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4477..4581
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4583..4940
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4583..4774
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4606..4626)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4791..4926
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4975..5769
                     /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             4975..5769
                     /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(5029..5048)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5639..5658
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5957..5976)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6001..6048
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6377..6965
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6866..6885
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgaggc caccatgaag atggacaaaa agactatagt ttggtttaga
      601 agagacctaa ggattgagga taatcctgca ttagcagcag ctgctcacga aggatctgtt
      661 tttcctgtct tcatttggtg tcctgaagaa gaaggacagt tttatcctgg aagagcttca
      721 agatggtgga tgaaacaatc acttgctcac ttatctcaat ccttgaaggc tcttggatct
      781 gacctcactt taatcaaaac ccacaacacg atttcagcga tcttggattg tatccgcgtt
      841 accggtgcta caaaagtcgt ctttaaccac ctctatgatc ctgtttcgtt agttcgggac
      901 cataccgtaa aggagaagct ggtggaacgt gggatctctg tgcaaagcta caatggagat
      961 ctattgtatg aaccgtggga gatatactgc gaaaagggca aaccttttac gagtttcaat
     1021 tcttactgga agaaatgctt agatatgtcg attgaatccg ttatgcttcc tcctccttgg
     1081 cggttgatgc caataactgc agcggctgaa gcgatttggg cgtgttcgat tgaagaacta
     1141 gggctggaga atgaggccga gaaaccgagc aatgcgttgt taactagagc ttggtctcca
     1201 ggatggagca atgctgataa gttactaaat gagttcatcg agaagcagtt gatagattat
     1261 gcaaagaaca gcaagaaagt tgttgggaat tctacttcac tactttctcc gtatctccat
     1321 ttcggggaaa taagcgtcag acacgttttc cagtgtgccc ggatgaaaca aattatatgg
     1381 gcaagagata agaacagtga aggagaagaa agtgcagatc tttttcttag gggaatcggt
     1441 ttaagagagt attctcggta tatatgtttc aacttcccgt ttactcacga gcaatcgttg
     1501 ttgagtcatc ttcggttttt cccttgggat gctgatgttg ataagttcaa ggcctggaga
     1561 caaggcagga ccggttatcc gttggtggat gccggaatga gagagctttg ggctaccgga
     1621 tggatgcata acagaataag agtgattgtt tcaagctttg ctgtgaagtt tcttctcctt
     1681 ccatggaaat ggggaatgaa gtatttctgg gatacacttt tggatgctga tttggaatgt
     1741 gacatccttg gctggcagta tatctctggg agtatccccg atggccacga gcttgatcgc
     1801 ttggacaatc ccgcgttaca aggcgccaaa tatgacccag aaggtgagta cataaggcaa
     1861 tggcttcccg agcttgcgag attgccaact gaatggatcc atcatccatg ggacgctcct
     1921 ttaaccgtac tcaaagcttc tggtgtggaa ctcggaacaa actatgcgaa acccattgta
     1981 gacatcgaca cagctcgtga gctactagct aaagctattt caagaacccg tgaagcacag
     2041 atcatgatcg gagcagcagc ccgggatcca ccggtcgcca ccatggtgag caagggcgag
     2101 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
     2161 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     2221 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
     2281 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
     2341 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     2401 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
     2461 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
     2521 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
     2581 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac
     2641 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc
     2701 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
     2761 tccaccggcg gcatggacga gctgtacaag ggttcttcta aatttgggat caaccgccct
     2821 gctgtgctgg cgccagagga tgggctggcc atgtccctac acttcatgac actgggtggc
     2881 agttctcttt cccctactga gggcaaaggc tccggactcc agggccacat catggagaac
     2941 ccacagtact tcagtgatac ctgtgtccac catatcaagc gccaggacat cattctcaag
     3001 tgggagctag gggagggagc ctttggaaag gtctttcttg ctgagtgcta caaccttctg
     3061 aatgatcagg acaagatgct ggtggctgtc aaggcactga aggagacatc tgagaatgct
     3121 cgtcaggact tccatcgtga ggcagagctg ctcaccatgc tacagcacca acacatcgta
     3181 cgcttctttg gagtctgcac ggagggtggc ccattgctca tggtcttcga gtacatgcgc
     3241 catggggacc tcaaccgttt cctccggtcc cacggacctg atgcaaaact gctggctggc
     3301 ggcgaggatg tggctcctgg tcctttgggc cttgggcagc ttctggctgt ggctagccag
     3361 gtggctgctg gtatggtgta cctagccagc ctgcactttg tgcaccggga cctggccaca
     3421 cgcaactgtc tggtgggtca gggactagtg gtcaagattg gagacttcgg catgagcagg
     3481 gacatctaca gcacagacta ctaccgtgtg ggaggtcgga ccatgctgcc catccgctgg
     3541 atgcctccag agagcatcct ctaccgcaag ttcagcaccg agagtgatgt gtggagcttc
     3601 ggggtggtgc tctgggagat cttcacctat ggaaagcaac cctggtacca gctctccaac
     3661 actgaggcga tcgagtgcat cacgcagggc cgggagctgg agcggccgcg cgcctgccct
     3721 cctgatgtct acgccatcat gcgcggctgc tggcagcgtg agccgcaaca gcgcctcagc
     3781 atgaaggatg tgcacgcgcg gctgcaggcc ttggcacagg cgccaccgag ttacctggac
     3841 gttctgggtt agcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3901 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3961 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     4021 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4081 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4141 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4201 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4261 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4321 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4381 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4441 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4501 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4561 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4621 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4681 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4741 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4801 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4861 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4921 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4981 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5041 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5101 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     5161 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5221 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5281 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5341 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5401 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5461 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     5521 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5581 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5641 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5701 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5761 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5821 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5881 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5941 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     6001 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     6061 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     6121 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     6181 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     6241 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6301 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6361 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6421 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6481 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6541 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6601 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6661 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6721 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6781 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6841 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6901 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6961 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7021 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt
     7081 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccg
//
