LOCUS       sequence-247882-        7550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses optoFGFR1 with mutation in PHR to suppress 
            homointeraction, thus cannot be activated by light.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7550)
  AUTHORS   Kim N, Kim JM, Lee M, Kim CY, Chang KY, Heo WD
  TITLE     Spatiotemporal control of fibroblast growth factor receptor signals 
            by blue light.
  JOURNAL   Chem Biol. 2014 Jul 17;21(7):903-12. doi: 
            10.1016/j.chembiol.2014.05.013. Epub 2014 Jun 26.
  PUBMED    24981772
REFERENCE   2  (bases 1 to 7550)
  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.chembiol.2014.05.013"; journalName: "Chem Biol"; date: 
            "2014-07-17- 17"; volume: "21"; issue: "7"; pages: "903-12"
COMMENT     Sequence Label: pCMV-OptoFGFR1-D387A
FEATURES             Location/Qualifiers
     source          1..7550
                     /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      535..544
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3430..3439
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3436..3438,3439..3441,3442..4152)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3481..3502)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3742..3761)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4089..4110
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4278..4399
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4315..4334)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4369..4388
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4406..4861)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4543..4564)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4755..4774
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4888..4992
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4994..5351
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4994..5185
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5017..5037)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5202..5337
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5386..6180
                     /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             5386..6180
                     /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(5440..5459)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6050..6069
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6368..6387)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6412..6459
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6788..7376
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7277..7296
                     /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 gctagccacc
      541 atgggatgta taaaatcaaa agggaaagac agcgcgggag cagatagtgc tggtagtgct
      601 ggtagtgctg ggaattccaa gatgaagagt ggtaccaaga agagtgactt ccacagccag
      661 atggctgtgc acaagctggc caagagcatc cctctgcgca gacaggtaac agtgtctgct
      721 gactccagtg catccatgaa ctctggggtt cttctggttc ggccatcacg gctctcctcc
      781 agtgggactc ccatgctagc aggggtctct gagtatgagc ttcccgaaga ccctcgctgg
      841 gagctgcctc gggacagact ggtcttaggc aaacccctgg gagagggctg ctttgggcag
      901 gtggtgttgg cagaggctat cgggctggac aaggacaaac ccaaccgtgt gaccaaagtg
      961 gctgtgaaga tgttgaagtc ggacgcaaca gagaaagact tgtcagacct gatctcagaa
     1021 atggagatga tgaagatgat cgggaagcat aagaatatca tcaacctgct gggggcctgc
     1081 acgcaggatg gtcccttgta tgtcatcgtg gagtatgcct ccaagggcaa cctgcgggag
     1141 tacctgcagg cccggaggcc cccagggctg gaatactgct acaaccccag ccacaaccca
     1201 gaggagcagc tctcctccaa ggacctggtg tcctgcgcct accaggtggc ccgaggcatg
     1261 gagtatctgg cctccaagaa gtgcatacac cgagacctgg cagccaggaa tgtcctggtg
     1321 acagaggaca atgtgatgaa gatagcagac tttggcctcg cacgggacat tcaccacatc
     1381 gactactata aaaagacaac caacggccga ctgcctgtga agtggatggc acccgaggca
     1441 ttatttgacc ggatctacac ccaccagagt gatgtgtggt ctttcggggt gctcctgtgg
     1501 gagatcttca ctctgggcgg ctccccatac cccggtgtgc ctgtggagga acttttcaag
     1561 ctgctgaagg agggtcaccg catggacaag cccagtaact gcaccaacga gctgtacatg
     1621 atgatgcggg actgctggca tgcagtgccc tcacagagac ccaccttcaa gcagctggtg
     1681 gaagacctgg accgcatcgt ggccttgacc tccaaccagg agtacctgga cctgtccatg
     1741 cccctggacc agtactcccc cagctttccc gacacccgga gctctacgtg ctcctcaggg
     1801 gaggattccg tcttctctca tgagccgctg cccgaggagc cctgcctgcc ccgacaccca
     1861 gcccagcttg ccaatggcgg actcaaacgc cgcgcggatc caccggtcgc caccatgaag
     1921 atggacaaaa agaccatcgt ctggtttcgg agagatttga gaatagaaga taatcccgcg
     1981 ctcgccgccg cggcccacga gggttccgtc ttccccgttt tcatttggtg tcctgaagaa
     2041 gaaggccagt tttatcccgg aagggcctct aggtggtgga tgaagcaaag tctggcccat
     2101 cttagccagt cactgaaagc actgggcagt gatcttaccc tgatcaagac acacaatacc
     2161 atctctgcca tcctcgactg catcagggtg accggcgcaa cgaaagtcgt gtttaaccac
     2221 ctgtacgatc cagttagtct ggtgcgcgac cacactgtga aggagaagct ggtggaacgg
     2281 gggatcagtg tgcagagcta caacggggac cttctgtacg agccatggga gatctattgc
     2341 gagaaaggga aaccgttcac ctccttcaac agttactgga agaaatgttt ggatatgtca
     2401 atagagtccg ttatgttgcc ccctccctgg agactgatgc cgattactgc tgctgcagag
     2461 gccatctggg cctgctccat cgaggaactc ggtctggaaa atgaagcaga aaagccaagc
     2521 aatgcacttc tcactagagc ctggagcccc ggctggtcta atgccgacaa gctgcttaac
     2581 gagttcatcg aaaaacaact gattgactac gcgaagaact ccaagaaagt ggtaggtaac
     2641 tcaactagct tgctctctcc atatctccat tttggcgaga tttctgtccg ccatgtattt
     2701 cagtgcgctc ggatgaaaca gattatctgg gctcgcgata aaaacagcga aggcgaagaa
     2761 agcgccgatc tgttcctgcg agggatcgga cttcgggaat actcccggta tatatgtttc
     2821 aactttccat tcacacacga gcagagtctg ttgtcccacc tcaggttctt cccctgggac
     2881 gccgatgtcg acaaattcaa ggcatggaga cagggaagga cagggtaccc actcgtggat
     2941 gctggcatga gagagctctg ggctacaggc tggatgcaca accgcatccg ggtaatcgtg
     3001 tcctcatttg ctgtcaagtt tctgctcctg ccttggaaat ggggaatgaa gtacttttgg
     3061 gatacccttc tcgccgccga cttggagtgt gacattctgg gatggcaata tattagcggg
     3121 tcaattcctg acggccatga gttggacagg ttggacaatc cggccttgca gggagctaag
     3181 tatgatcccg aaggagagta tattcgacag tggctccccg agctggcccg acttcctacg
     3241 gagtggattc accatccttg ggacgcacca ctgacagtgc tcaaggcaag cggggtggag
     3301 ctgggcacca attacgctaa gcctatagtt gatatagata cagcacgcga gctgctggct
     3361 aaagcgatct ctcgcactcg ggaggcgcag attatgatcg gtgctgccgg cagcggcagc
     3421 ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3481 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     3541 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     3601 gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgatgtgctt tgcccgctac
     3661 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     3721 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     3781 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     3841 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     3901 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     3961 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     4021 ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag
     4081 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     4141 gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     4201 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     4261 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4321 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4381 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     4441 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     4501 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     4561 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     4621 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     4681 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4741 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     4801 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     4861 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4921 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4981 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     5041 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5101 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5161 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     5221 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5281 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5341 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     5401 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     5461 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5521 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     5581 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5641 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5701 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5761 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5821 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5881 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     5941 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     6001 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     6061 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     6121 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     6181 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     6241 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     6301 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     6361 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     6421 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     6481 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     6541 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     6601 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     6661 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6721 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6781 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6841 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6901 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6961 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7021 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7081 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7141 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7201 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7261 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7321 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7381 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     7441 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
     7501 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
