LOCUS       sequence-208646-        7214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of Lyn-tagged intracellular domain of TrkA (aa 
            450-799) fused to mCherry and CRY2PHR (aa 1-498).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7214)
  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 7214)
  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: Lyn-iTrkA-mCh-CRY2
FEATURES             Location/Qualifiers
     source          1..7214
                     /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"
     regulatory      538..547
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1681..1690
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1687..2394
                     /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(1832..1850)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2100..2119
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2431..3924
                     /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"
     polyA_signal    3942..4063
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3979..3998)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4033..4052
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4070..4525)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4207..4228)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4419..4438
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4552..4656
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4658..5015
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4658..4849
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4681..4701)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4866..5001
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5050..5844
                     /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             5050..5844
                     /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(5104..5123)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5714..5733
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6032..6051)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6076..6123
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6452..7040
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6941..6960
                     /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 gctagcggcc
      541 accatgggat gtattaaatc aaaaaggaaa gacaatctca atgacgatct cgagctcaag
      601 cttcgaattc tgcagtcgac gaaatttggg atcaaccgcc ctgctgtgct ggcgccagag
      661 gatgggctgg ccatgtccct acacttcatg acactgggtg gcagttctct ttcccctact
      721 gagggcaaag gctccggact ccagggccac atcatggaga acccacagta cttcagtgat
      781 acctgtgtcc accatatcaa gcgccaggac atcattctca agtgggagct aggggaggga
      841 gcctttggaa aggtctttct tgctgagtgc tacaaccttc tgaatgatca ggacaagatg
      901 ctggtggctg tcaaggcact gaaggagaca tctgagaatg ctcgtcagga cttccatcgt
      961 gaggcagagc tgctcaccat gctacagcac caacacatcg tacgcttctt tggagtctgc
     1021 acggagggtg gcccattgct catggtcttc gagtacatgc gccatgggga cctcaaccgt
     1081 ttcctccggt cccacggacc tgatgcaaaa ctgctggctg gcggcgagga tgtggctcct
     1141 ggtcctttgg gccttgggca gcttctggct gtggctagcc aggtggctgc tggtatggtg
     1201 tacctagcca gcctgcactt tgtgcaccgg gacctggcca cacgcaactg tctggtgggt
     1261 cagggactag tggtcaagat tggagacttc ggcatgagca gggacatcta cagcacagac
     1321 tactaccgtg tgggaggtcg gaccatgctg cccatccgct ggatgcctcc agagagcatc
     1381 ctctaccgca agttcagcac cgagagtgat gtgtggagct tcggggtggt gctctgggag
     1441 atcttcacct atggaaagca accctggtac cagctctcca acactgaggc gatcgagtgc
     1501 atcacgcagg gccgggagct ggagcggccg cgcgcctgcc ctcctgatgt ctacgccatc
     1561 atgcgcggct gctggcagcg tgagccgcaa cagcgcctca gcatgaagga tgtgcacgcg
     1621 cggctgcagg ccttggcaca ggcgccaccg agttacctgg acgttctggg tccaccggtc
     1681 gccaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     1741 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     1801 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     1861 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     1921 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     1981 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     2041 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     2101 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     2161 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     2221 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     2281 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     2341 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagggcagt
     2401 agcggaagca gtggttcttc aggatcatcc atgaagatgg acaaaaagac tatagtttgg
     2461 tttagaagag acctaaggat tgaggataat cctgcattag cagcagctgc tcacgaagga
     2521 tctgtttttc ctgtcttcat ttggtgtcct gaagaagaag gacagtttta tcctggaaga
     2581 gcttcaagat ggtggatgaa acaatcactt gctcacttat ctcaatcctt gaaggctctt
     2641 ggatctgacc tcactttaat caaaacccac aacacgattt cagcgatctt ggattgtatc
     2701 cgcgttaccg gtgctacaaa agtcgtcttt aaccacctct atgatcctgt ttcgttagtt
     2761 cgggaccata ccgtaaagga gaagctggtg gaacgtggga tctctgtgca aagctacaat
     2821 ggagatctat tgtatgaacc gtgggagata tactgcgaaa agggcaaacc ttttacgagt
     2881 ttcaattctt actggaagaa atgcttagat atgtcgattg aatccgttat gcttcctcct
     2941 ccttggcggt tgatgccaat aactgcagcg gctgaagcga tttgggcgtg ttcgattgaa
     3001 gaactagggc tggagaatga ggccgagaaa ccgagcaatg cgttgttaac tagagcttgg
     3061 tctccaggat ggagcaatgc tgataagtta ctaaatgagt tcatcgagaa gcagttgata
     3121 gattatgcaa agaacagcaa gaaagttgtt gggaattcta cttcactact ttctccgtat
     3181 ctccatttcg gggaaataag cgtcagacac gttttccagt gtgcccggat gaaacaaatt
     3241 atatgggcaa gagataagaa cagtgaagga gaagaaagtg cagatctttt tcttagggga
     3301 atcggtttaa gagagtattc tcggtatata tgtttcaact tcccgtttac tcacgagcaa
     3361 tcgttgttga gtcatcttcg gtttttccct tgggatgctg atgttgataa gttcaaggcc
     3421 tggagacaag gcaggaccgg ttatccgttg gtggatgccg gaatgagaga gctttgggct
     3481 accggatgga tgcataacag aataagagtg attgtttcaa gctttgctgt gaagtttctt
     3541 ctccttccat ggaaatgggg aatgaagtat ttctgggata cacttttgga tgctgatttg
     3601 gaatgtgaca tccttggctg gcagtatatc tctgggagta tccccgatgg ccacgagctt
     3661 gatcgcttgg acaatcccgc gttacaaggc gccaaatatg acccagaagg tgagtacata
     3721 aggcaatggc ttcccgagct tgcgagattg ccaactgaat ggatccatca tccatgggac
     3781 gctcctttaa ccgtactcaa agcttctggt gtggaactcg gaacaaacta tgcgaaaccc
     3841 attgtagaca tcgacacagc tcgtgagcta ctagctaaag ctatttcaag aacccgtgaa
     3901 gcacagatca tgatcggagc agcataacaa ttgttgttgt taacttgttt attgcagctt
     3961 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4021 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     4081 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     4141 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     4201 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     4261 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     4321 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4381 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4441 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4501 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4561 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4621 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     4681 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4741 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4801 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4861 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4921 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     4981 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     5041 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     5101 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     5161 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     5221 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5281 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5341 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5401 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5461 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5521 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     5581 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5641 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5701 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5761 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5821 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5881 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5941 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     6001 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     6061 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     6121 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     6181 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     6241 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     6301 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6361 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6421 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6481 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6541 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6601 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6661 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6721 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6781 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6841 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6901 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6961 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7021 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7081 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     7141 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     7201 tatatggagt tccg
//
