LOCUS       sequence-415568-        7280 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses DRD2-V2tail-TevN-BLITz-TetR-FKBP component in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7280)
  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 7280)
  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: DRD2-V2tail-TevN-BLITz-TetR-FKBP
FEATURES             Location/Qualifiers
     source          1..7280
                     /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"
     CDS             2470..2496
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             3310..3330
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3331..3948
                     /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(3401..3420)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             3955..4278
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     primer_bind     complement(4287..4306)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4288..4306)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    4311..4445
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4335..4354)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4389..4408
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4558..4578)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4558..4576)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4597..4613)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4597..4613)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4610..4632)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4621..4637
                     /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(4645..4651,4652..4669,4670..4675))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4690..4711
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4828..4845)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4998..5586)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5078..5097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5757..6548,6549..6617))
                     /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     6380..6399
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6618..6722)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6748..7203)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6885..6906)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7097..7116
                     /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 acttgttctt tttgcaatga agacgatcat cgccctgagc
     1081 tacatcttct gcctggtatt cgccgactac aaggacgatg atgacgccag catggatcca
     1141 ctgaacctgt cctggtatga tgatgatctg gagaggcaga actggagccg gcccttcaat
     1201 gggtcagaag ggaaggcaga caggccccac tacaactact atgccatgct gctcaccctc
     1261 ctcatcttta tcatcgtctt tggcaatgtg ctggtgtgca tggctgtatc ccgagagaag
     1321 gctttgcaga ccaccaccaa ctacttgata gtcagccttg ctgtggctga tcttctggtg
     1381 gccacactgg taatgccgtg ggttgtctac ctggaggtgg tgggtgagtg gaaattcagc
     1441 aggattcact gtgacatctt tgtcactctg gatgtcatga tgtgcacagc aagcatcctg
     1501 aacctgtgtg ccatcagcat tgacaggtac acagctgtgg caatgcccat gctgtataac
     1561 acacgctaca gctccaagcg ccgagttact gtcatgattg ccattgtctg ggtcctgtcc
     1621 ttcaccatct cctgcccact gctcttcgga ctcaacaata cagaccagaa tgagtgtatc
     1681 attgccaacc ctgcctttgt ggtctactcc tccattgtct cattctacgt gcccttcatc
     1741 gtcactctgc tggtctatat caaaatctac atcgtcctcc ggaagcgccg gaagcgggtc
     1801 aacaccaagc gcagcagtcg agctttcaga gccaacctga agacaccact caaggatgct
     1861 gcccgccgag ctcaggagct ggaaatggag atgctgtcaa gcaccagccc cccagagagg
     1921 acccggtata gccccatccc tcccagtcac caccagctca ctctccctga tccatcccac
     1981 cacggcctac atagcaaccc tgacagtcct gccaaaccag agaagaatgg gcacgccaag
     2041 attgtcaatc ccaggattgc caagttcttt gagatccaga ccatgcccaa tggcaaaacc
     2101 cggacctccc ttaagacgat gagccgcaga aagctctccc agcagaagga gaagaaagcc
     2161 actcagatgc ttgccattgt tctcggtgtg ttcatcatct gctggctgcc cttcttcatc
     2221 acgcacatcc tgaatataca ctgtgattgc aacatcccac cagtcctcta cagcgccttc
     2281 acatggctgg gctatgtcaa cagtgccgtc aaccccatca tctacaccac cttcaacatc
     2341 gagttccgca aggccttcat gaagatcttg cactgcgaat tcggacgcac cccacccagc
     2401 ctgggtcccc aagatgagtc ctgcaccacc gccagctcct ccctggccaa ggacacttca
     2461 tcgcttcaac ttcctcctct tgaacgtctt actctcggtg gctctggagg tctcgagatg
     2521 ggcgagagcc tgttcaaggg acctagggac tacaacccta tcagtagcac aatttgtcac
     2581 ctgaccaacg agagtgatgg ccacacaaca agcctgtacg gcatcgggtt cggacccttt
     2641 atcatcacca acaagcacct gttcaggcgg aataatggca ctctgctggt gcagagcctg
     2701 cacggggtgt tcaaagtgaa gaacacaacc actctgcagc agcacctgat cgatgggcgg
     2761 gatatgatca tcattaggat gcccaaggac ttcccccctt ttcctcagaa actgaagttc
     2821 cgagagcccc agagagagga gagaatctgt ctggtgacca caaactttca ggagctcggt
     2881 agtggtagtg gggagtttct ggcaaccaca ctggaacgga tcgagaaaaa tttcgtgatt
     2941 actgatccga gactgcctga caacccaatc atttttgcga gcgattcctt cctgcagctg
     3001 acagaatatt ctcgggaaga gatcctgggg cgcaattgcc gttttctgca gggacccgag
     3061 acagaccgtg ccactgttcg gaaaatcaga gatgctattg acaaccagac tgaagtgacc
     3121 gttcagctga tcaattatac caagagcggc aagaagttct ggaacgtgtt ccacctgcag
     3181 ccgatgcgcg attataaggg cgacgtccag tacttcattg gcgtgcagct ggatggcacc
     3241 gaacgtcttc atggcgccgc tgagcgtgag gcggtctgcc tgatcaaaaa gacagccttt
     3301 cagattgctg agaacctgta cttccagggc tctagattag ataaaagtaa agtgattaac
     3361 agcgcattag agctgcttaa tgaggtcgga atcgaaggtt taacaacccg taaactcgcc
     3421 cagaagctag gtgtagagca gcctacattg tattggcatg taaaaaataa gcgggctttg
     3481 ctcgacgcct tagccattga gatgttagat aggcaccata ctcacttttg ccctttagaa
     3541 ggggaaagct ggcaagattt tttacgtaat aacgctaaaa gttttagatg tgctttacta
     3601 agtcatcgcg atggagcaaa agtacattta ggtacacggc ctacagaaaa acagtatgaa
     3661 actctcgaaa atcaattagc ctttttatgc caacaaggtt tttcactaga gaatgcatta
     3721 tatgcactca gcgctgtggg gcattttact ttaggttgcg tattggaaga tcaagagcat
     3781 caagtcgcta aagaagaaag ggaaacacct actactgata gtatgccgcc attattacga
     3841 caagctatcg aattatttga tcaccaaggt gcagagccag ccttcttatt cggccttgaa
     3901 ttgatcatat gcggattaga aaaacaactt aaatgtgaaa gtgggtccac cggtggagtg
     3961 caggtggaaa ccatctcccc aggagacggg cgcaccttcc ccaagcgcgg ccagacctgc
     4021 gtggtgcact acaccgggat gcttgaagat ggaaagaaat ttgattcctc ccgggacaga
     4081 aacaagccct ttaagtttat gctaggcaag caggaggtga tccgaggctg ggaagaaggg
     4141 gttgcccaga tgagtgtggg tcagagagcc aaactgacta tatctccaga ttatgcctat
     4201 ggtgccactg ggcacccagg catcatccca ccacatgcca ctctcgtctt cgatgtggag
     4261 cttctaaaac tggaataggc tagcctccct atagtgagtc gtattacgta gatccagaca
     4321 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     4381 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     4441 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     4501 ttttttaatt cgcgtccgcg gcgccaatgc attgggcccg gtacccagct tttgttccct
     4561 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4621 ttgttatccg ctcacaattc cacacaacat acgagccggg agcataaagt gtaaagcctg
     4681 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4741 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4801 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4861 gctgcgggag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4921 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4981 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5041 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5101 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5161 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5221 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5281 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5341 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5401 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5461 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5521 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5581 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5641 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5701 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5761 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5821 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5881 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5941 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6001 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6061 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6121 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6181 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6241 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6301 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6361 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6421 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6481 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6541 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6601 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6661 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6721 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     6781 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     6841 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     6901 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     6961 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     7021 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     7081 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     7141 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     7201 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     7261 cgctattacg ccagtcgacc
//
