LOCUS       sequence-436957-        6889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PDGFR-Cry2olig-FRB-P2A-GFP(1-10) component in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6889)
  AUTHORS   Lee S, Cui M, Lee D, Han K, Sun W, Lee D
  TITLE     A rationally designed optochemogenetic switch for activating 
            canonical Wnt signaling.
  JOURNAL   iScience. 2023 Feb 19;26(3):106233. doi: 10.1016/j.isci.2023.106233.
            eCollection 2023 Mar 17.
  PUBMED    36915690
REFERENCE   2  (bases 1 to 6889)
  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"; journalName: "iScience.";
            date: "2023-02-19"; volume: "26(3):106233. doi"; pages: " 
            10.1016/j.isci.2023.106233. eCollection 2023 Mar 17"
COMMENT     Sequence Label: pCMV:: PDGFR-Cry2olig-FRB-P2A-GFP(1-10)
FEATURES             Location/Qualifiers
     source          1..6889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     sig_peptide     1050..1109
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             1113..1142
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1146..1292
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             1320..2813
                     /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
                     SGVELGTNYAKPIVDIDTARELLAKAISRTRGAQIMIGAA"
     CDS             2853..3131
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     CDS             3147..3203
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3222..3863
                     /codon_start=1
                     /product="fragment of superfolder GFP consisting of the 
                     first 10 beta-strands"
                     /label=GFP(1-10)
                     /note="associates with GFP11, the 11th beta-strand, to 
                     reconstitute fluorescent GFP (Cabantous et al., 2005; 
                     Kamiyama et al., 2016)"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATIGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     KYKTRAVVKFEGDTLVNRIELKGTDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFTVRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQTVLSKDPNEK"
     primer_bind     complement(3896..3915)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3897..3915)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3920..4054
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3944..3963)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3998..4017
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4167..4187)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4167..4185)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4206..4222)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4206..4222)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4219..4241)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4230..4246
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4254..4260,4261..4278,4279..4284))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4299..4320
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4437..4454)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4607..5195)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4687..4706)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5366..6157,6158..6226))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5989..6008
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6227..6331)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6357..6812)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6494..6515)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6706..6725
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttcta tggagacaga cacactcctg ctatgggtac
     1081 tgctgctctg ggttccaggt tccactggtg acgaacaaaa actcatctca gaagaggatc
     1141 tgaatgctgt gggccaggac acgcaggagg tcatcgtggt gccacactcc ttgcccttta
     1201 aggtggtggt gatctcagcc atcctggccc tggtggtgct caccatcatc tcccttatca
     1261 tcctcatcat gctttggcag aagaagccac gtggtggctc tggaggtctc gaggccacca
     1321 tgaagatgga caaaaagact atagtttggt ttagaagaga cctaaggatt gaggataatc
     1381 ctgcattagc agcagctgct cacgaaggat ctgtttttcc tgtcttcatt tggtgtcctg
     1441 aagaagaagg acagttttat cctggaagag cttcaagatg gtggatgaaa caatcacttg
     1501 ctcacttatc tcaatccttg aaggctcttg gatctgacct cactttaatc aaaacccaca
     1561 acacgatttc agcgatcttg gattgtatcc gcgttaccgg tgctacaaaa gtcgtcttta
     1621 accacctcta tgatcctgtt tcgttagttc gggaccatac cgtaaaggag aagctggtgg
     1681 aacgtgggat ctctgtgcaa agctacaatg gagatctatt gtatgaaccg tgggagatat
     1741 actgcgaaaa gggcaaacct tttacgagtt tcaattctta ctggaagaaa tgcttagata
     1801 tgtcgattga atccgttatg cttcctcctc cttggcggtt gatgccaata actgcagcgg
     1861 ctgaagcgat ttgggcgtgt tcgattgaag aactagggct ggagaatgag gccgagaaac
     1921 cgagcaatgc gttgttaact agagcttggt ctccaggatg gagcaatgct gataagttac
     1981 taaatgagtt catcgagaag cagttgatag attatgcaaa gaacagcaag aaagttgttg
     2041 ggaatagtac ttcactactt tctccgtatc tccatttcgg ggaaataagc gtcagacacg
     2101 ttttccagtg tgcccggatg aaacaaatta tatgggcaag agataagaac agtgaaggag
     2161 aagaaagtgc agatcttttt cttaggggaa tcggtttaag agagtattct cggtatatat
     2221 gtttcaactt cccgtttact cacgagcaat cgttgttgag tcatcttcgg tttttccctt
     2281 gggatgctga tgttgataag ttcaaggcct ggagacaagg caggaccggt tatccgttgg
     2341 tggatgccgg aatgagagag ctttgggcta ccggatggat gcataacaga ataagagtga
     2401 ttgtttcaag ctttgctgtg aagtttcttc tccttccatg gaaatgggga atgaagtatt
     2461 tctgggatac acttttggat gctgatttgg aatgtgacat ccttggctgg cagtatatct
     2521 ctgggagtat ccccgatggc cacgagcttg atcgcttgga caatcccgcg ttacaaggcg
     2581 ccaaatatga cccagaaggt gagtacataa ggcaatggct tcccgagctt gcgagattgc
     2641 caactgaatg gattcatcat ccatgggacg ctcctttaac cgtactcaaa gcttctggtg
     2701 tggaactcgg aacaaactat gcgaaaccca ttgtagacat cgacacagct cgtgagctac
     2761 tagctaaagc tatttcaaga acccgtggag cacagatcat gatcggagca gcactgcagg
     2821 agctcggtgg aggcggccgc atggcttctc gaatcctctg gcatgagatg tggcatgaag
     2881 gcctggaaga ggcatctcgt ttgtactttg gggaaaggaa cgtgaaaggc atgtttgagg
     2941 tgctggagcc cttgcatgct atgatggaac ggggccccca gactctgaag gaaacatcct
     3001 ttaatcaggc ctatggtcga gatttaatgg aggcccaaga gtggtgcagg aagtacatga
     3061 aatcagggaa tgtcaaggac ctcctccaag cctgggacct ctattatcat gtgttccgac
     3121 gaatctcaaa gactagtgga tccggcgcta ctaacttcag cctgctgaag caggctggag
     3181 acgtggagga gaaccctgga cctgaattcg ttaacagatc tatgtccaaa ggagaagaac
     3241 tgtttaccgg tgttgtgcca attttggttg aactcgatgg tgatgtcaac ggacataagt
     3301 tctcagtgag aggcgaagga gaaggtgacg ccaccattgg aaaattgact cttaaattca
     3361 tctgtactac tggtaaactt cctgtaccat ggccgactct cgtaacaacg cttacgtacg
     3421 gagttcagtg cttttcgaga tacccagacc atatgaaaag acatgacttt tttaagtcgg
     3481 ctatgcctga aggttacgtg caagaaagaa caatttcgtt caaagatgat ggaaaatata
     3541 aaactagagc agttgttaaa tttgaaggag atactttggt taaccgcatt gaactgaaag
     3601 gaacagattt taaagaagat ggtaatattc ttggacacaa actcgaatac aattttaata
     3661 gtcataacgt atacatcact gctgataagc aaaagaacgg aattaaagcg aatttcacag
     3721 tacgccataa tgtagaagat ggcagtgttc aacttgccga ccattaccaa caaaacaccc
     3781 ctattggaga cggtccggta cttcttcctg ataatcacta cctctcaaca caaacagtcc
     3841 tgagcaaaga tccaaatgaa aaaggaacag gtggcggcgg aagttagtct agcctcccta
     3901 tagtgagtcg tattacgtag atccagacat gataagatac attgatgagt ttggacaaac
     3961 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     4021 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     4081 gtttcaggtt cagggggagg tgtgggaggt tttttaattc gcgtccgcgg cgccaatgca
     4141 ttgggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     4201 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4261 cgagccggga gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4321 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4381 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4441 ctcactgact cgctgcgctc ggtcgttcgg ctgcgggagc ggtatcagct cactcaaagg
     4501 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4561 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4621 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4681 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4741 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4801 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4861 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4921 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4981 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5041 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5101 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5161 agcagcagat tatgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5221 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5281 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5341 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5401 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5461 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5521 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5581 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5641 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5701 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5761 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5821 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5881 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5941 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6001 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6061 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6121 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6181 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6241 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6301 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
     6361 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     6421 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     6481 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     6541 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     6601 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     6661 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     6721 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     6781 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg
     6841 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgaca
//
