LOCUS       sequence-393108-        6507 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the GAL4 DNA Binding domain linked to the FKBP12(F36V) 
            target protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6507)
  AUTHORS   Ma D, Yuan Q, Peng F, Paredes V, Zeng H, Osikpa EC, Yang Q, Peddi A,
            Patel A, Liu MS, Sun Z, Gao X
  TITLE     Engineered PROTAC-CID Systems for Mammalian Inducible Gene 
            Regulation.
  JOURNAL   J Am Chem Soc. 2023 Jan 25;145(3):1593-1606. doi: 
            10.1021/jacs.2c09129. Epub 2023 Jan 10.
  PUBMED    36626587
REFERENCE   2  (bases 1 to 6507)
  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: 
            "10.1021/jacs.2c09129"; journalName: "J Am Chem Soc"; date: 
            "2023-01-25- 25"; volume: "145"; issue: "3"; pages: "1593-1606"
COMMENT     Sequence Label: PC3-709-33-10_GAL4-FKBP12(F36V)
FEATURES             Location/Qualifiers
     source          1..6507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     107..126
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        201..305
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(306..374,375..1166)
                     /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     complement(524..543)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1337..1925
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1826..1845
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2064..2089
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    2761..2781
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     2848..2870
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2862..2879
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2863..2879
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2886..2905
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2886..2904
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        3014..3317
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          3595..4614
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4538..4560
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4622..4641
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    4672..4696
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4736..4756
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4787..5227
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             join(5240..5242,5243..5563)
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             5582..5629
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    complement(5664..5688)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5778..5797)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5843..5898
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5897..5916)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6259..6275)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6259..6275)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6272..6294)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6283..6299
                     /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(6307..6313,6314..6331,6332..6337))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6352..6373
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cagggtaatt cggtcaagcc ttgccttgtt gtagcttaaa ttttgctcgc gcactactca
       61 gcgacctcca acacacaagc agggagcaga tactggctta actatgcggc atcagagcag
      121 attgtactga gagtgcacca taggggatcg ggagatctcc cgatccgtcg acgtcaggtg
      181 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
      241 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      301 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      361 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      421 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      481 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      541 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      601 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      661 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      721 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      781 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      841 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      901 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      961 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     1021 ggtcacgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     1081 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1141 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1201 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1261 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1321 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1381 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1441 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1501 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1561 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1621 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1681 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1741 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1801 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1861 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1921 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     1981 acatgttctt tattaccctg ttatccctaa ccggtttacc gagctcttat tggttttcaa
     2041 acttcattga ctgtgccaag cttctgacct cttctcttcc tcccacaggg cctcgagaca
     2101 attgcatcga tggtaccagt gaaagtctgc aatgaattcg agttggcgcg cctgtcattc
     2161 taaatctctc tttcagccta aagctttttc cccgtatccc cccaggtgtc tgcaggctca
     2221 aagagcagcg agaagcgttc agaggaaagc gatcccgtgc caccttcccc gtgcccgggc
     2281 tgtccccgca cgctgccggc tcggggatgc ggggggagcg ccggaccgga gcggagcccc
     2341 gggcggctcg ctgctgcccc ctagcggggg agggacgtaa ttacatccct gggggctttg
     2401 ggagggggct gtccccgtga gctcccaggc gcgcctgtca ttctaaatct ctctttcagc
     2461 ctaaagcttt ttccccgtat ccccccaggt gtctgcaggc tcaaagagca gcgagaagcg
     2521 ttcagaggaa agcgatcccg tgccaccttc cccgtgcccg ggctgtcccc gcacgctgcc
     2581 ggctcgggga tgcgggggga gcgccggacc ggagcggagc cccgggcggc tcgctgctgc
     2641 cccctagcgg gggagggacg taattacatc cctgggggct ttgggggggg gctgtccccg
     2701 tgagctccca ggcgcgccaa ctcggtaccg tatcgatgtc gacgttaacg ctagtgatat
     2761 caactttgta tagaaaagtt ggctccgaat ttctcgagtt aattaaatgt gctgcaaggc
     2821 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     2881 aattgtaata cgactcacta tagggcgaat tggagctagg cctacgtagc gcgcgagctc
     2941 caccgcggtg gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     3001 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     3061 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     3121 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     3181 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     3241 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     3301 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
     3361 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
     3421 gggcgggggg ggggggggcg cgcgccaggc ggggsggggs ggggsgrggg gsggggsggg
     3481 gsgrggsggr gaggkgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
     3541 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
     3601 gagtcgctgc gttgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc
     3661 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
     3721 ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc
     3781 cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg
     3841 tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc
     3901 gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc
     3961 ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgggcg gggtgtgtgc
     4021 gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc
     4081 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc
     4141 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     4201 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     4261 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     4321 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     4381 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     4441 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     4501 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     4561 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     4621 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcga cccaagtttg
     4681 tacaaaaaag caggctgatt accggagaat tggcggccgc accggtgcca ccatgccaaa
     4741 gaagaagcgg aaggtcggtg gcggctcacc cgggggtggc ggctcaatga agctactgtc
     4801 ttctatcgaa caagcatgcg atatttgccg acttaaaaag ctcaagtgct ccaaagaaaa
     4861 accgaagtgc gccaagtgtc tgaagaacaa ctgggagtgt cgctactctc ccaaaaccaa
     4921 aaggtcgccg ctgactaggg cacatctgac agaagtggaa tcaaggctag aaagactgga
     4981 acagctattt ctactgattt ttcctcgaga agaccttgac atgattttga aaatggattc
     5041 tttacaggat ataaaagcat tgttaacagg attatttgta caagataatg tgaataaaga
     5101 tgccgtcaca gatagattgg cttcagtgga gactgatatg cctctaacat tgagacagca
     5161 tagaataagt gcgacatcat catcggaaga gagtagtaac aaaggtcaaa gacagttgac
     5221 tgtatcgggt ggcggctcaa tgggagtgca ggtggaaacc atctccccag gagacgggcg
     5281 caccttcccc aagcgcggcc agacctgcgt ggtgcactac accgggatgc ttgaagatgg
     5341 aaagaaagtt gattcctccc gggacagaaa caagcccttt aagtttatgc taggcaagca
     5401 ggaggtgatc cgaggctggg aagaaggggt tgcccagatg agtgtgggtc agagagccaa
     5461 actgactata tctccagatt atgcctatgg tgccactggg cacccaggca tcatcccacc
     5521 acatgccact ctcgtcttcg atgtggagct tctaaaactg gaaactagtg gtggcggctc
     5581 aaagcgtcct gctgctacta agaaagctgg tcaagctaag aaaaagaaat ctagatagtg
     5641 actcgagact agataactga tctacccagc tttcttgtac aaagtggtac gcgtgaattc
     5701 actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct
     5761 cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc
     5821 cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg
     5881 aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag
     5941 aatgagtatt tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa
     6001 ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc
     6061 catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt
     6121 tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc
     6181 tgactactcc cagtcatagc tgtccctctt ctcttatgga gatccctcga cctgcagccc
     6241 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6301 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6361 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6421 ccagcggatc gacagtacta agcttggtgc gtttttatgc ttgtagtatt gtataatgtt
     6481 tttaagatcc ttaattaata gggataa
//
