LOCUS       sequence-169199-        5829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fusion of PTEN with a modified human FKBP variant harboring the F36V
            and L106P point mutations (FKBP*) leads to degradation of the fusion
            protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5829)
  AUTHORS   An W, Jackson RE, Hunter P, Gogel S, van Diepen M, Liu K, Meyer MP, 
            Eickholt BJ
  TITLE     Engineering FKBP-Based Destabilizing Domains to Build Sophisticated 
            Protein Regulation Systems.
  JOURNAL   PLoS One. 2015 Dec 30;10(12):e0145783. doi: 
            10.1371/journal.pone.0145783. eCollection 2015.
  PUBMED    26717575
REFERENCE   2  (bases 1 to 5829)
  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: "PLoS One.";
            date: "2015-12-30"; volume: "10(12):e0145783. doi"; pages: " 
            10.1371/journal.pone.0145783. eCollection 2015"
COMMENT     Sequence Label: FKBP*-PTEN
FEATURES             Location/Qualifiers
     source          1..5829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        368..645
                     /label=chicken beta-actin promoter
     intron          646..1662
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1586..1608
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1670..1689
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1724..1733
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1730..1732,1733..2053)
                     /codon_start=1
                     /product="destabilization domain that can be stabilized by 
                     Shield1 in the ProteoTuner(TM) system"
                     /label=DD
                     /note="L106P mutant of FKBP12"
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLSSGKKVDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKPE"
     primer_bind     complement(3364..3383)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3429..3484
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3483..3502)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3890..3907)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4061..4649)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4141..4160)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4820..5611,5612..5680))
                     /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     5443..5462
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5681..5785)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(5816..5829,1..366)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca
      421 cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg
      481 gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg
      541 agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg
      601 cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc
      661 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
      721 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
      781 cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc
      841 cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg
      901 gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg
      961 gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg
     1021 tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga
     1081 gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt
     1141 gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc
     1201 gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc
     1261 ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg
     1321 gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg
     1381 tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc
     1441 ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg
     1501 ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc
     1561 cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga
     1621 gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg
     1681 gttattgtgc tgtctcatca ttttggcaaa gaattcgaag cttgccacca tgggagtgca
     1741 ggtggaaacc atctccccag gagacgggcg caccttcccc aagcgcggcc agacctgtgt
     1801 ggtgcactac accgggatgc ttagcagcgg aaagaaagtc gattcctccc gggacagaaa
     1861 caagcccttt aagtttatgc taggcaagca ggaggtgatc cgaggctggg aagaaggggt
     1921 tgcccagatg agtgtgggtc agagagccaa actgactata tctccagatt atgcctatgg
     1981 tgccactggg cacccaggca tcatcccacc acatgccact ctcgtcttcg atgtggagct
     2041 tctaaaaccg gaaggatcct ctgggggtgg cggtccgtgg aggcgaagcg gcggtggagg
     2101 ttctggtggt ggaggtggca cagccatcat caaagagatc gttagcagaa acaaaaggag
     2161 atatcaagag gatggattcg acttagactt gacctatatt tatccaaaca ttattgctat
     2221 gggatttcct gcagaaagac ttgaaggcgt atacaggaac aatattgatg atgtagtaag
     2281 gtttttggat tcaaagcata aaaaccatta caagatatac aatctttgtg ctgaaagaca
     2341 ttatgacacc gccaaattta attgcagagt tgcacaatat ccttttgaag accataaccc
     2401 accacagcta gaacttatca aacccttttg tgaagatctt gaccaatggc taagtgaaga
     2461 tgacaatcat gttgcagcaa ttcactgtaa agctggaaag ggacgaactg gtgtaatgat
     2521 atgtgcatat ttattacatc ggggcaaatt tttaaaggca caagaggccc tagatttcta
     2581 tggggaagta aggaccagag acaaaaaggg agtaactatt cccagtcaga ggcgctatgt
     2641 gtattattat agctacctgt taaagaatca tctggattat agaccagtgg cactgttgtt
     2701 tcacaagatg atgtttgaaa ctattccaat gttcagtggc ggaacttgca atcctcagtt
     2761 tgtggtctgc cagctaaagg tgaagatata ttcctccaat tcaggaccca cacgacggga
     2821 agacaagttc atgtactttg agttccctca gccgttacct gtgtgtggtg atatcaaagt
     2881 agagttcttc cacaaacaga acaagatgct aaaaaaggac aaaatgtttc acttttgggt
     2941 aaatacattc ttcataccag gaccagagga aacctcagaa aaagtagaaa atggaagtct
     3001 atgtgatcaa gaaatcgata gcatttgcag tatagagcgt gcagataatg acaaggaata
     3061 tctagtactt actttaacaa aaaatgatct tgacaaagca aataaagaca aagccaaccg
     3121 atacttttct ccaaatttta aggtgaagct gtacttcaca aaaacagtag aggagccgtc
     3181 aaatccagag gctagcagtt caacttctgt aacaccagat gttagtgaca atgaacctga
     3241 tcattataga tattctgaca ccactgactc tgatccagag aatgaacctt ttgatgaaga
     3301 tcagcataca caaattacaa aagtctgata agcggccgct ggccagctag ctaggtagct
     3361 agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac
     3421 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     3481 ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt
     3541 ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag
     3601 aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt
     3661 gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct
     3721 aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt
     3781 catagctgtc cctcttctct tatggagatc cctcgaccga tccgctgcat taatgaatcg
     3841 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3901 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3961 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4021 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4081 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4141 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4201 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4261 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4321 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4381 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4441 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4501 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4561 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4621 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4681 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4741 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4801 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4861 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4921 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4981 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5041 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5101 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5161 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5221 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5281 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5341 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5401 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5461 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5521 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5581 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5641 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5701 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5761 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gggtcgacat
     5821 tgattattg
//
