LOCUS       sequence-415577-        6900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses 
            PDGFR-KA2-TevCs-43LK-iLID(139)-NES-NES-TevN-iLID-TEVseq-TetR-FKBP 
            component in rat hippocampal neuron cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6900)
  AUTHORS   Cui M, Lee S, Ban SH, Ryu JR, Shen M, Yang SH, Kim JY, Choi SK, Han 
            J, Kim Y, Han K, Lee D, Sun W, Kwon HB, Lee D
  TITLE     A single-component, light-assisted uncaging switch for 
            endoproteolytic release.
  JOURNAL   Nat Chem Biol. 2024 Mar;20(3):353-364. doi: 
            10.1038/s41589-023-01480-6. Epub 2023 Nov 16.
  PUBMED    37973890
REFERENCE   2  (bases 1 to 6900)
  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: 
            "10.1038/s41589-023-01480-6"; journalName: "Nat Chem Biol"; date: 
            "2024-03"; volume: "20"; issue: "3"; pages: "353-364"
COMMENT     Sequence Label: 
            hSyn-PDGFR-KA2-TevCs-43LK-iLID(139)-NES-NES-TevN-iLID-TEVseq-TetR-FK
            BP
FEATURES             Location/Qualifiers
     source          1..6900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..455
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     promoter        464..482
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     464..481
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     sig_peptide     501..560
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             564..593
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             597..743
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             774..1223
                     /codon_start=1
                     /product="first 150 residues of the mouse kainate receptor 
                     subunit 2"
                     /label=KA2
                     /note="mediates targeting to the neuronal soma (Shemesh et 
                     al., 2017)"
                     /translation="MPAELLLLLIVAFANPSCQVLSSLRMAAILDDQTVCGRGERLALA
                     LAREQINGIIEVPAKARVEVDIFELQRDSQYETTDTMCQILPKGVVSVLGPSSSPASAS
                     TVSHICGEKEIPHIKVGPEETPRLQYLRFASVSLYPSNEDVSLAVS"
     CDS             2100..2126
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             2136..2162
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             2937..2957
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2958..3575
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="SRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVK
                     NKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRP
                     TEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTD
                     SMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(3028..3047)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             3582..3905
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     primer_bind     complement(3914..3933)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3915..3933)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3938..4072
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3962..3981)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4016..4035
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4185..4205)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4185..4203)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4224..4240)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4224..4240)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4237..4259)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4248..4264
                     /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(4272..4278,4279..4296,4297..4302))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4317..4338
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4455..4472)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4625..5213)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4705..4724)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5384..6175,6176..6244))
                     /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     6007..6026
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6245..6349)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6375..6830)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     6724..6743
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtcgactagt gcaagtgggt tttaggacca ggatgaggcg gggtgggggt gcctacctga
       61 cgaccgaccc cgacccactg gacaagcacc caacccccat tccccaaatt gcgcatcccc
      121 tatcagagag ggggagggga aacaggatgc ggcgaggcgc gtgcgcactg ccagcttcag
      181 caccgcggac agtgccttcg cccccgcctg gcggcgcgcg ccaccgccgc ctcagcactg
      241 aaggcgcgct gacgtcactc gccggtcccc cgcaaactcc ccttcccggc caccttggtc
      301 gcgtccgcgc cgccgccggc ccagccggac cgcaccacgc gaggcgcgag ataggggggc
      361 acgggcgcga ccatctgcgc tgcggcgccg gcgactcagc gctgcctcag tctgcggtgg
      421 gcagcggagg agtcgtgtcg tgcctgagag cgcagaagct ttcatttagg tgacactata
      481 gaatacaagc tacttgttct atggagacag acacactcct gctatgggta ctgctgctct
      541 gggttccagg ttccactggt gacgaacaaa aactcatctc agaagaggat ctgaatgctg
      601 tgggccagga cacgcaggag gtcatcgtgg tgccacactc cttgcccttt aaggtggtgg
      661 tgatctcagc catcctggcc ctggtggtgc tcaccatcat ctcccttatc atcctcatca
      721 tgctttggca gaagaagcca cgtggtggct ctggaggtct cgagggaggt accatgcccg
      781 ccgaactgct gctgctgctg attgtcgctt ttgctaaccc ttcttgccag gtgctgtctt
      841 ctctgaggat ggccgctatt ctggacgatc agaccgtgtg cggaagagga gagaggctgg
      901 cactggcact ggctagggag cagatcaatg gcatcattga agtgcctgca aaggcccggg
      961 tcgaggtgga cattttcgaa ctgcagagag atagccagta cgagaccaca gacaccatgt
     1021 gccagatcct gccaaaagga gtggtctccg tcctgggacc aagctcctct cctgcttctg
     1081 caagtaccgt gtctcacatt tgtggcgaga aggaaatccc ccatatcaag gtcgggccag
     1141 aggaaacacc caggctgcag tacctgcgct tcgcctcagt gagcctgtat ccatcaaacg
     1201 aggatgtgtc actggcagtc agcggaggta gcctcgagaa gtccatgtcc agcatggtga
     1261 gcgatactag ctgtaccttc ccatcatctg acggaatctt ctggaagcac tggattcaga
     1321 ctaaggacgg ccagtgtggc agcccactgg tgagcacacg agacggattc atcgtgggga
     1381 ttcacagcgc ctccaacttt acaaacacca ataactattt cacctcagtg ccaaagaact
     1441 ttatggagct gctgaccaac caggaggccc agcagtgggt gagcgggtgg cgcctgaacg
     1501 ccgattccgt gctgtggggc gggcacaagg tgtttatgac cgcatgcggt ggcggtggct
     1561 ctggaggtgg tgggtccgga ggaggaggtg gcggttccgg cggaggtggg tctggtggag
     1621 gtggatctgg aggtggtgga tcaggaggag gaggttcagg cggccgcgag tttctggcaa
     1681 ccacactgga acggatcgag aaaaatttcg tgattactga tccgagactg cctgacaacc
     1741 caatcatttt tgcgagcgat tccttcctgc aactgacaga atattctcgg gaagagatcc
     1801 tggggcgcaa ttgccgtttt ctgcaaggac ccgagacaga ccgtgccact gttcggaaaa
     1861 tcagagatgc tattgacaac cagactgaag tgaccgttca gctgatcaat tataccaaga
     1921 gcggcaagaa gttctggaac gtgttccacc tgcaaccgat gcgcgattat aagggcgacg
     1981 tccagtactt cattggcgtg cagctggatg gcaccgaacg tcttcatggc gccgctgagc
     2041 gtgaggcggt ctgcctgatc aaaaagacag cctttcagat tgctggatcc atgcttcaac
     2101 ttcctcctct tgaacgtctt actctcatgt tgcagctacc accattagag agactaacac
     2161 tgatgggcga gagcctgttc aagggaccta gggactacaa ccctatcagt agcacaattt
     2221 gtcacctgac caacgagagt gatggccaca caacaagcct gtacggcatc gggttcggac
     2281 cctttatcat caccaacaag cacctgttca ggcggaataa tggcactctg ctggtgcaga
     2341 gcctgcacgg ggtgttcaaa gtgaagaaca caaccactct gcagcagcac ctgatcgatg
     2401 ggcgggatat gatcatcatt aggatgccca aggacttccc cccttttcct cagaaactga
     2461 agttccgaga gccccagaga gaggagagaa tctgtctggt gaccacaaac tttcagaccg
     2521 agtttctggc aaccacactg gaacggatcg agaaaaattt cgtgattact gatccgagac
     2581 tgcctgacaa cccaatcatt tttgcgagcg attccttcct gcagctgaca gaatattctc
     2641 gggaagagat cctggggcgc aattgccgtt ttctgcaggg acccgagaca gaccgtgcca
     2701 ctgttcggaa aatcagagat gctattgaca accagactga agtgaccgtt cagctgatca
     2761 attataccaa gagcggcaag aagttctgga acgtgttcca cctgcagccg atgcgcgatt
     2821 ataagggcga cgtccagtac ttcattggcg tgcagctgga tggcaccgaa cgtcttcatg
     2881 gcgccgctga gcgtgaggcg gtctgcctga tcaaaaagac agcctttcag attgctgaga
     2941 acctgtactt ccagggctct agattagata aaagtaaagt gattaacagc gcattagagc
     3001 tgcttaatga ggtcggaatc gaaggtttaa caacccgtaa actcgcccag aagctaggtg
     3061 tagagcagcc tacattgtat tggcatgtaa aaaataagcg ggctttgctc gacgccttag
     3121 ccattgagat gttagatagg caccatactc acttttgccc tttagaaggg gaaagctggc
     3181 aagatttttt acgtaataac gctaaaagtt ttagatgtgc tttactaagt catcgcgatg
     3241 gagcaaaagt acatttaggt acacggccta cagaaaaaca gtatgaaact ctcgaaaatc
     3301 aattagcctt tttatgccaa caaggttttt cactagagaa tgcattatat gcactcagcg
     3361 ctgtggggca ttttacttta ggttgcgtat tggaagatca agagcatcaa gtcgctaaag
     3421 aagaaaggga aacacctact actgatagta tgccgccatt attacgacaa gctatcgaat
     3481 tatttgatca ccaaggtgca gagccagcct tcttattcgg ccttgaattg atcatatgcg
     3541 gattagaaaa acaacttaaa tgtgaaagtg ggtccaccgg tggagtgcag gtggaaacca
     3601 tctccccagg agacgggcgc accttcccca agcgcggcca gacctgcgtg gtgcactaca
     3661 ccgggatgct tgaagatgga aagaaatttg attcctcccg ggacagaaac aagcccttta
     3721 agtttatgct aggcaagcag gaggtgatcc gaggctggga agaaggggtt gcccagatga
     3781 gtgtgggtca gagagccaaa ctgactatat ctccagatta tgcctatggt gccactgggc
     3841 acccaggcat catcccacca catgccactc tcgtcttcga tgtggagctt ctaaaactgg
     3901 aataggctag cctccctata gtgagtcgta ttacgtagat ccagacatga taagatacat
     3961 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     4021 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     4081 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc
     4141 gtccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta
     4201 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4261 acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga
     4321 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4381 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4441 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcgggagcgg
     4501 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4561 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4621 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4681 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4741 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4801 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4861 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4921 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4981 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5041 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5101 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5161 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5221 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5281 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5341 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5401 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5461 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5521 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5581 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5641 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5701 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5761 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5821 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5881 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5941 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6001 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6061 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6121 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6181 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6241 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6301 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6361 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     6421 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     6481 agaatagacc gagatagggt tgagtgttgt tccggtttgg aacaagagtc cactattaaa
     6541 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     6601 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     6661 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     6721 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     6781 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
     6841 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
//
