LOCUS       sequence-415567-        7040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses LAUNCHER component in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7040)
  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 7040)
  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: 
            pCMV-PDGFR-TevCs-43LK-iLID(139)-NES-NES-TevN-iLID-TEVseq-tTA
FEATURES             Location/Qualifiers
     source          1..7040
                     /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        1014..1032
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1014..1031
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     sig_peptide     1051..1110
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             1114..1143
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1147..1293
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             2176..2202
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             2212..2238
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             3013..3033
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(3034..3651,3652..4038)
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus 
                     VP16"
                     /label=tTA
                     /note="In the Tet-Off(R) system, tTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="SRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVK
                     NKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRP
                     TEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTD
                     SMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTIE
                     GLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHAD
                     ALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(3104..3123)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(4047..4066)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4048..4066)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    4071..4205
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4095..4114)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4149..4168
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4318..4338)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4318..4336)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4357..4373)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4357..4373)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4370..4392)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4381..4397
                     /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(4405..4411,4412..4429,4430..4435))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4450..4471
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4588..4605)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4758..5346)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4838..4857)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5517..6308,6309..6377))
                     /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     6140..6159
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6378..6482)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6508..6963)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     6857..6876
                     /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 ttcatttagg
     1021 tgacactata gaatacaagc tacttgttct atggagacag acacactcct gctatgggta
     1081 ctgctgctct gggttccagg ttccactggt gacgaacaaa aactcatctc agaagaggat
     1141 ctgaatgctg tgggccagga cacgcaggag gtcatcgtgg tgccacactc cttgcccttt
     1201 aaggtggtgg tgatctcagc catcctggcc ctggtggtgc tcaccatcat ctcccttatc
     1261 atcctcatca tgctttggca gaagaagcca cgtggtggct ctggaggact cgagaagtcc
     1321 atgtccagca tggtgagcga tactagctgt accttcccat catctgacgg aatcttctgg
     1381 aagcactgga ttcagactaa ggacggccag tgtggcagcc cactggtgag cacacgagac
     1441 ggattcatcg tggggattca cagcgcctcc aactttacaa acaccaataa ctatttcacc
     1501 tcagtgccaa agaactttat ggagctgctg accaaccagg aggcccagca gtgggtgagc
     1561 gggtggcgcc tgaacgccga ttccgtgctg tggggcgggc acaaggtgtt tatgaccgca
     1621 tgcggtggcg gtggctctgg aggtggtggg tccggaggag gaggtggcgg ttccggcgga
     1681 ggtgggtctg gtggaggtgg atctggaggt ggtggatcag gaggaggagg ttcaggcggc
     1741 cgcgagtttc tggcaaccac actggaacgg atcgagaaaa atttcgtgat tactgatccg
     1801 agactgcctg acaacccaat catttttgcg agcgattcct tcctgcaact gacagaatat
     1861 tctcgggaag agatcctggg gcgcaattgc cgttttctgc aaggacccga gacagaccgt
     1921 gccactgttc ggaaaatcag agatgctatt gacaaccaga ctgaagtgac cgttcagctg
     1981 atcaattata ccaagagcgg caagaagttc tggaacgtgt tccacctgca gccgatgcgc
     2041 gattataagg gcgacgtcca gtacttcatt ggcgtgcagc tggatggcac cgaacgtctt
     2101 catggcgccg ctgagcgtga ggcggtctgc ctgatcaaaa agacagcctt tcagattgct
     2161 ggatccatgc ttcaacttcc tcctcttgaa cgtcttactc tcatgttgca gctaccacca
     2221 ttagagagac taacactgat gggcgagagc ctgttcaagg gacctaggga ctacaaccct
     2281 atcagtagca caatttgtca cctgaccaac gagagtgatg gccacacaac aagcctgtac
     2341 ggcatcgggt tcggaccctt tatcatcacc aacaagcacc tgttcaggcg gaataatggc
     2401 actctgctgg tgcagagcct gcacggggtg ttcaaagtga agaacacaac cactctgcag
     2461 cagcacctga tcgatgggcg ggatatgatc atcattagga tgcccaagga cttcccccct
     2521 tttcctcaga aactgaagtt ccgagagccc cagagagagg agagaatctg tctggtgacc
     2581 acaaactttc agaccgagtt tctggcaacc acactggaac ggatcgagaa aaatttcgtg
     2641 attactgatc cgagactgcc tgacaaccca atcatttttg cgagcgattc cttcctgcag
     2701 ctgacagaat attctcggga agagatcctg gggcgcaatt gccgttttct gcagggaccc
     2761 gagacagacc gtgccactgt tcggaaaatc agagatgcta ttgacaacca gactgaagtg
     2821 accgttcagc tgatcaatta taccaagagc ggcaagaagt tctggaacgt gttccacctg
     2881 cagccgatgc gcgattataa gggcgacgtc cagtacttca ttggcgtgca gctggatggc
     2941 accgaacgtc ttcatggcgc cgctgagcgt gaggcggtct gcctgatcaa aaagacagcc
     3001 tttcagattg ctgagaacct gtacttccag ggctctagat tagataaaag taaagtgatt
     3061 aacagcgcat tagagctgct taatgaggtc ggaatcgaag gtttaacaac ccgtaaactc
     3121 gcccagaagc taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa taagcgggct
     3181 ttgctcgacg ccttagccat tgagatgtta gataggcacc atactcactt ttgcccttta
     3241 gaaggggaaa gctggcaaga ttttttacgt aataacgcta aaagttttag atgtgcttta
     3301 ctaagtcatc gcgatggagc aaaagtacat ttaggtacac ggcctacaga aaaacagtat
     3361 gaaactctcg aaaatcaatt agccttttta tgccaacaag gtttttcact agagaatgca
     3421 ttatatgcac tcagcgctgt ggggcatttt actttaggtt gcgtattgga agatcaagag
     3481 catcaagtcg ctaaagaaga aagggaaaca cctactactg atagtatgcc gccattatta
     3541 cgacaagcta tcgaattatt tgatcaccaa ggtgcagagc cagccttctt attcggcctt
     3601 gaattgatca tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc cgcgtacagc
     3661 cgcgcgcgta cgaaaaacaa ttacgggtct accatcgagg gcctgctcga tctcccggac
     3721 gacgacgccc ccgaagaggc ggggctggcg gctccgcgcc tgtcctttct ccccgcggga
     3781 cacacgcgca gactgtcgac ggcccccccg accgatgtca gcctggggga cgagctccac
     3841 ttagacggcg aggacgtggc gatggcgcat gccgacgcgc tagacgattt cgatctggac
     3901 atgttggggg acggggattc cccgggtccg ggatttaccc cccacgactc cgccccctac
     3961 ggcgctctgg atatggccga cttcgagttt gagcagatgt ttaccgatgc ccttggaatt
     4021 gacgagtacg gtgggtaggc tagcctccct atagtgagtc gtattacgta gatccagaca
     4081 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     4141 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     4201 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     4261 ttttttaatt cgcgtccgcg gcgccaatgc attgggcccg gtacccagct tttgttccct
     4321 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4381 ttgttatccg ctcacaattc cacacaacat acgagccggg agcataaagt gtaaagcctg
     4441 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4501 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4561 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4621 gctgcgggag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4681 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4741 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4801 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4861 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4921 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4981 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5041 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5101 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5161 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5221 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5281 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5341 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5401 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5461 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5521 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5581 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5641 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5701 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5761 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5821 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5881 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5941 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6001 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6061 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6121 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6181 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6241 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6301 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6361 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6421 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6481 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     6541 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     6601 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccggtt tggaacaaga
     6661 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     6721 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     6781 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     6841 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     6901 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     6961 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     7021 cgctattacg ccagtcgacc
//
