LOCUS       sequence-339453-        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)-UbK0G75S-sfGFP
FEATURES             Location/Qualifiers
     source          1..7568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             198..667
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    732..1089
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1154..1570
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1500..1522
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1538..1554
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1580..1954
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1892..1908
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1963..1972
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1969..1971,1972..2292)
                     /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             2299..2523
                     /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
                     GRQLEDGRTLSDYNIQRESTLHLVLRLRSG"
     CDS             join(2524..2526,2527..2529,2530..3243)
                     /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             2524..3240
                     /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(2557..2585)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3180..3200
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    join(3274..3848,3849..3851,3852..3860)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3441..3458)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3668..3687
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3861..4571
                     /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(4006..4024)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4274..4293
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     LTR             4742..5211
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5341..5358)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5512..6100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5592..5611)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(6224..6292,6293..7084)
                     /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(6442..6461)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcaggatcaa tttcccatta tgccagtcat attaattact agtcaattag ttgattttta
       61 tttttgacat atacatgtga aagaccccac ctgtaggttt ggcaagctag cttaagtaac
      121 gccattttgc aaggcatgga aaaatacata actgagaata gaaaagttca gatcaaggtc
      181 aggaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
      241 ccccggctca gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt
      301 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag
      361 ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg
      421 accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc
      481 tgctccccga gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat
      541 tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt
      601 gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg
      661 tctttcattt gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac
      721 caccgggagg taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg
      781 actgatttta tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc
      841 cgtggtggaa ctgacgagtt cggaacaccc ggccgcaacc ctgggagacg tcccatggac
      901 ttcgggggcc gtttttgtgg cccgacctga gtccaaaaat cccgatcgtt ttggactctt
      961 tggtgcaccc cccttagagg agggatatgt ggttctggta ggagacgaga acctaaaaca
     1021 gttcccgcct ccgtctgaat ttttgctttc ggtttgggac cgaagccgcg ccgcgcgtct
     1081 tgtctgctgc agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt atttgtctga
     1141 aaatatgggc ccgggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa
     1201 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac
     1261 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
     1321 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
     1381 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
     1441 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
     1501 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
     1561 tctaggcgcc cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa
     1621 cttccctgac cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca
     1681 ggctctctac ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca
     1741 actggaccga ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg
     1801 acaccagact aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac
     1861 ccccaccgcc ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc
     1921 tgccgacccc gggggtggac catcctctag actgccggat ccgccaccat gggagtgcag
     1981 gtggaaacca tctccccagg agacgggcgc accttcccca agcgcggcca gacctgtgtg
     2041 gtgcactaca ccgggatgct tgaagatgga aagaaagtcg attcctcccg ggacagaaac
     2101 aagcccttta agtttatgct aggcaagcag gaggtgatcc gaggctggga agaaggggtt
     2161 gcccagatga gtgtgggtca gagagccaaa ctgactatat ctccagatta tgcctatggt
     2221 gccactgggc acccaggcat catcccacca catgccactc tcgtcttcga tgtggagctt
     2281 ctaaaaccgg aaacgcgtca gatcttcgtc agaacgttaa ccggtagaac cataactcta
     2341 gaagttgaac catccgatac catcgaaaac gttagagcta gaattcaaga cagagaaggc
     2401 attccacctg atcaacaaag attgatcttt gccggtagac agctcgagga cggtagaacg
     2461 ctgtctgatt acaacattca gagagagtcg accttacatc ttgtcttaag actaagatcc
     2521 ggtatgcata gcaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta
     2581 gatggtgatg ttaatgggca caaattttct gtccgtggag agggtgaagg tgatgctaca
     2641 aacggaaaac tcacccttaa atttatttgc actactggaa aactacctgt tccgtggcca
     2701 acacttgtca ctactctgac ctatggtgtt caatgctttt cccgttatcc ggatcacatg
     2761 aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga acgcactata
     2821 tctttcaaag atgacgggac ctacaagacg cgagctgaag tcaagtttga aggtgatacc
     2881 cttgttaatc gtatcgagtt aaagggtatt gattttaaag aagatggaaa cattcttgga
     2941 cacaaactcg agtacaactt taactcacac aatgtataca tcacggcaga caaacaaaag
     3001 aatggaatca aagctaactt caaaattcgc cacaacgttg aagatggttc cgttcaacta
     3061 gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac
     3121 cattacctgt cgacacaatc tgtcctttcg aaagatccca acgaaaagcg tgaccacatg
     3181 gtccttcttg agtttgtaac tgctgctggg attacacatg gcatggatga gctctacaaa
     3241 taagcggccg ctacgtaaat tccgcccccc ccccccctct ccctcccccc cccctaacgt
     3301 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac
     3361 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
     3421 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa
     3481 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
     3541 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
     3601 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
     3661 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc
     3721 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag
     3781 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     3841 atgataatat ggccacaacc atgatgagca agggcgagga ggataacatg gccatcatca
     3901 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga
     3961 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     4021 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct
     4081 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg
     4141 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc
     4201 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
     4261 tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc
     4321 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg
     4381 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc
     4441 tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca
     4501 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
     4561 tgtacaagta agtcgacgat aaaataaaag attttattta gtctccagaa aaagggggga
     4621 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
     4681 tggaaaaata cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca
     4741 gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     4801 agaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     4861 cccggctcag ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta
     4921 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
     4981 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     5041 taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg
     5101 gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc
     5161 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc acatgcagca
     5221 tgtatcaaaa ttaatttggt tttttttctt aagtatttac attaaatggc catagttgca
     5281 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5341 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5401 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5461 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5521 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5581 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5641 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5701 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5761 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5821 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5881 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5941 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6001 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6061 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6121 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagaca
     6181 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     6241 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6301 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     6361 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     6421 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     6481 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     6541 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     6601 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     6661 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     6721 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     6781 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     6841 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6901 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6961 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     7021 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     7081 gtaactgtca gaccaagttt actcatatat actttagatt gatttgcggc cggccgcaaa
     7141 aggatctagg tgaagatcct ttttgataat ctcatgagcg gatacatatt tgaatgtatt
     7201 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc accagctttg
     7261 ctcttaggag tttcctaata catcccaaac tcaaatatat aaagcatttg acttgttcta
     7321 tgccctaggg ggcgggggga agctaagcca gcttttttta acatttaaaa tgttaattcc
     7381 attttaaatg cacagatgtt tttatttcat aagggtttca atgtgcatga atgctgcaat
     7441 attcctgtta ccaaagctag tataaataaa aatagataaa cgtggaaatt acttagagtt
     7501 tctgtcatta acgtttcctt cctcagttga caacataaat gcgctgctga gaagccagtt
     7561 tgcatctg
//
