LOCUS       sequence-339426-        7568 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression for LIBRON constructs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7568)
  AUTHORS   Miyamae Y, Chen LC, Utsugi Y, Farrants H, Wandless TJ
  TITLE     A Method for Conditional Regulation of Protein Stability in Native 
            or Near-Native Form.
  JOURNAL   Cell Chem Biol. 2020 Dec 17;27(12):1573-1581.e3. doi: 
            10.1016/j.chembiol.2020.09.004. Epub 2020 Oct 1.
  PUBMED    33007216
REFERENCE   2  (bases 1 to 7568)
  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: 
            "10.1016/j.chembiol.2020.09.004"; journalName: "Cell Chem Biol"; 
            date: "2020-12-17- 17"; volume: "27"; issue: "12"; pages: 
            "1573-1581.e3"
COMMENT     Sequence Label: pBMN-FKBP DD(L106P)-UbK0G75C-sfGFP
FEATURES             Location/Qualifiers
     source          1..7568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             121..590
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    655..1012
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1077..1493
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1423..1445
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1461..1477
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1503..1877
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1815..1831
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1886..1895
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1892..1894,1895..2215)
                     /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="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKPE"
     CDS             2222..2446
                     /codon_start=1
                     /product="human ubiquitin variant in which all of the 
                     lysines have been mutated to arginine"
                     /label=ubiquitin-K0
                     /note="also known as ubiquitin-KO"
                     /translation="QIFVRTLTGRTITLEVEPSDTIENVRARIQDREGIPPDQQRLIFA
                     GRQLEDGRTLSDYNIQRESTLHLVLRLRCG"
     CDS             join(2447..2449,2450..2452,2453..3166)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MHSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             2447..3163
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MHSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(2480..2508)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3103..3123
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    join(3197..3771,3772..3774,3775..3783)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3364..3381)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3591..3610
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3784..4494
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MMSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3929..3947)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4197..4216
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     LTR             4665..5134
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5264..5281)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5435..6023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5515..5534)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(6147..6215,6216..7007)
                     /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(6365..6384)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca gatggaacag
      121 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      241 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
      301 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
      361 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
      421 aaaagagccc acaacccctc actcggcgcg ccagtcctcc gattgactga gtcgcccggg
      481 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
      541 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
      601 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggtaagctg
      661 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
      721 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
      781 gttcggaaca cccggccgca accctgggag acgtcccatg gacttcgggg gccgtttttg
      841 tggcccgacc tgagtccaaa aatcccgatc gttttggact ctttggtgca ccccccttag
      901 aggagggata tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg
      961 aatttttgct ttcggtttgg gaccgaagcc gcgccgcgcg tcttgtctgc tgcagcatcg
     1021 ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaatatg ggcccgggcc
     1081 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
     1141 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
     1201 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
     1261 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
     1321 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
     1381 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
     1441 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gcccccatat
     1501 ggccatatga gatcttatat ggggcacccc cgccccttgt aaacttccct gaccctgaca
     1561 tgacaagagt tactaacagc ccctctctcc aagctcactt acaggctctc tacttagtcc
     1621 agcacgaagt ctggagacct ctggcggcag cctaccaaga acaactggac cgaccggtgg
     1681 tacctcaccc ttaccgagtc ggcgacacag tgtgggtccg ccgacaccag actaagaacc
     1741 tagaacctcg ctggaaagga ccttacacag tcctgctgac cacccccacc gccctcaaag
     1801 tagacggcat cgcagcttgg atacacgccg cccacgtgaa ggctgccgac cccgggggtg
     1861 gaccatcctc tagactgccg gatccgccac catgggagtg caggtggaaa ccatctcccc
     1921 aggagacggg cgcaccttcc ccaagcgcgg ccagacctgt gtggtgcact acaccgggat
     1981 gcttgaagat ggaaagaaag tcgattcctc ccgggacaga aacaagccct ttaagtttat
     2041 gctaggcaag caggaggtga tccgaggctg ggaagaaggg gttgcccaga tgagtgtggg
     2101 tcagagagcc aaactgacta tatctccaga ttatgcctat ggtgccactg ggcacccagg
     2161 catcatccca ccacatgcca ctctcgtctt cgatgtggag cttctaaaac cggaaacgcg
     2221 tcagatcttc gtcagaacgt taaccggtag aaccataact ctagaagttg aaccatccga
     2281 taccatcgaa aacgttagag ctagaattca agacagagaa ggcattccac ctgatcaaca
     2341 aagattgatc tttgccggta gacagctcga ggacggtaga acgctgtctg attacaacat
     2401 tcagagagag tcgaccttac atcttgtctt aagactaaga tgtggtatgc atagcaaagg
     2461 agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg
     2521 gcacaaattt tctgtccgtg gagagggtga aggtgatgct acaaacggaa aactcaccct
     2581 taaatttatt tgcactactg gaaaactacc tgttccgtgg ccaacacttg tcactactct
     2641 gacctatggt gttcaatgct tttcccgtta tccggatcac atgaaacggc atgacttttt
     2701 caagagtgcc atgcccgaag gttatgtaca ggaacgcact atatctttca aagatgacgg
     2761 gacctacaag acgcgagctg aagtcaagtt tgaaggtgat acccttgtta atcgtatcga
     2821 gttaaagggt attgatttta aagaagatgg aaacattctt ggacacaaac tcgagtacaa
     2881 ctttaactca cacaatgtat acatcacggc agacaaacaa aagaatggaa tcaaagctaa
     2941 cttcaaaatt cgccacaacg ttgaagatgg ttccgttcaa ctagcagacc attatcaaca
     3001 aaatactcca attggcgatg gccctgtcct tttaccagac aaccattacc tgtcgacaca
     3061 atctgtcctt tcgaaagatc ccaacgaaaa gcgtgaccac atggtccttc ttgagtttgt
     3121 aactgctgct gggattacac atggcatgga tgagctctac aaataagcgg ccgctacgta
     3181 aattccgccc cccccccccc tctccctccc ccccccctaa cgttactggc cgaagccgct
     3241 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     3301 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     3361 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     3421 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     3481 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     3541 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     3601 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     3661 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     3721 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca
     3781 accatgatga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     3841 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     3901 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     3961 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     4021 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     4081 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     4141 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     4201 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     4261 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     4321 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     4381 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     4441 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaagtcgac
     4501 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg
     4561 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact
     4621 gagaatagag aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa
     4681 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc
     4741 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     4801 aacagatggt ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt
     4861 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
     4921 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
     4981 cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat
     5041 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct
     5101 gagtgattga ctacccgtca gcgggggtct ttcacatgca gcatgtatca aaattaattt
     5161 ggtttttttt cttaagtatt tacattaaat ggccatagtt gcattaatga atcggccaac
     5221 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5281 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5341 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5401 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5461 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5521 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5581 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5641 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5701 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5761 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5821 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5881 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5941 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6001 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6061 agtggaacga aaactcacgt taagggattt tggtcatgag acaataaccc tgataaatgc
     6121 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     6181 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     6241 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     6301 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     6361 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     6421 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     6481 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6541 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     6601 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6661 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     6721 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     6781 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6841 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6901 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6961 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     7021 tttactcata tatactttag attgatttgc ggccggccgc aaaaggatct aggtgaagat
     7081 cctttttgat aatctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7141 aggggttccg cgcacatttc cccgaaaagt gccaccagct ttgctcttag gagtttccta
     7201 atacatccca aactcaaata tataaagcat ttgacttgtt ctatgcccta gggggcgggg
     7261 ggaagctaag ccagcttttt ttaacattta aaatgttaat tccattttaa atgcacagat
     7321 gtttttattt cataagggtt tcaatgtgca tgaatgctgc aatattcctg ttaccaaagc
     7381 tagtataaat aaaaatagat aaacgtggaa attacttaga gtttctgtca ttaacgtttc
     7441 cttcctcagt tgacaacata aatgcgctgc tgagaagcca gtttgcatct gtcaggatca
     7501 atttcccatt atgccagtca tattaattac tagtcaatta gttgattttt atttttgaca
     7561 tatacatg
//
