LOCUS Exported 8067 bp ds-DNA circular SYN 08-NOV-2022 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBMN FKBP(DD)-YFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8067) AUTHORS Banaszynski LA, Chen LC, Maynard-Smith LA, Ooi AG, Wandless TJ TITLE A rapid, reversible, and tunable method to regulate protein function in living cells using synthetic small molecules. JOURNAL Cell. 2006 Sep 8;126(5):995-1004. PUBMED 16959577 REFERENCE 2 (bases 1 to 8067) AUTHORS . TITLE Direct Submission JOURNAL Exported Nov 8, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8067 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 867..1336 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 1401..1758 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 1823..2239 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 2169..2191 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 2207..2223 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" misc_feature 2249..2623 /label=pol region /note="Moloney murine leukemia virus (MMLV) pol region containing the splice acceptor site" primer_bind 2561..2577 /label=pBMN 5' /note="MMLV sequence, for inserts in pBMN retroviral vector" regulatory 2632..2641 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2638..2961 /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 2968..3681 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(3010..3031) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3271..3290) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3618..3639 /label=EGFP-C /note="EGFP, forward primer" CDS 3688..3714 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" misc_feature 3771..4356 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(3937..3954) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4164..4183 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 4357..5040 /codon_start=1 /product="far-red fluorescent variant of Heteractis crispa chromoprotein" /label=HcRed1 /note="mammalian codon-optimized" /translation="MVSGLLKESMRIKMYMEGTVNGHYFKCEGEGDGNPFAGTQSMRIH VTEGAPLPFAFDILAPCCEYGSRTFVHHTAEIPDFFKQSFPEGFTWERTTTYEDGGILT AHQDTSLEGNCLIYKVKVHGTNFPADGPVMKNKSGGWEPSTEVVYPENGVLCGRNVMAL KVGDRHLICHHYTSYRSKKAVRALTMPGFHFTDIRLQMLRKKKDEYFELYEASVARYSD LPEKAN" CDS 5053..5739 /codon_start=1 /product="far-red fluorescent variant of Heteractis crispa chromoprotein" /label=HcRed1 /note="mammalian codon-optimized" /translation="MVSGLLKESMRIKMYMEGTVNGHYFKCEGEGDGNPFAGTQSMRIH VTEGAPLPFAFDILAPCCEYGSRTFVHHTAEIPDFFKQSFPEGFTWERTTTYEDGGILT AHQDTSLEGNCLIYKVKVHGTNFPADGPVMKNKSGGWEPSTEVVYPENGVLCGRNVMAL KVGDRHLICHHYTSYRSKKAVRALTMPGFHFTDIRLQMLRKKKDEYFELYEASVARYSD LPEKAN" LTR 5910..6379 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(6509..6526) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6680..7268) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6760..6779) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 7392..185 /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(7610..7629) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag 61 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg 121 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt 181 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttgcgg ccggccgcaa 241 aaggatctag gtgaagatcc tttttgataa tctcatgagc ggatacatat ttgaatgtat 301 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc caccagcttt 361 gctcttagga gtttcctaat acatcccaaa ctcaaatata taaagcattt gacttgttct 421 atgccctagg gggcgggggg aagctaagcc agcttttttt aacatttaaa atgttaattc 481 cattttaaat gcacagatgt ttttatttca taagggtttc aatgtgcatg aatgctgcaa 541 tattcctgtt accaaagcta gtataaataa aaatagataa acgtggaaat tacttagagt 601 ttctgtcatt aacgtttcct tcctcagttg acaacataaa tgcgctgctg agaagccagt 661 ttgcatctgt caggatcaat ttcccattat gccagtcata ttaattacta gtcaattagt 721 tgatttttat ttttgacata tacatgtgaa agaccccacc tgtaggtttg gcaagctagc 781 ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag aaaagttcag 841 atcaaggtca ggaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag 901 cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag 961 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat 1021 gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga 1081 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc 1141 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag 1201 tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc 1261 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt 1321 cagcgggggt ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca 1381 ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc cgattgtcta 1441 gtgtctatga ctgattttat gcgcctgcgt cggtactagt tagctaacta gctctgtatc 1501 tggcggaccc gtggtggaac tgacgagttc ggaacacccg gccgcaaccc tgggagacgt 1561 cccagggact tcgggggccg tttttgtggc ccgacctgag tccaaaaatc ccgatcgttt 1621 tggactcttt ggtgcacccc ccttagagga gggatatgtg gttctggtag gagacgagaa 1681 cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg gtttgggacc gaagccgcgc 1741 cgcgcgtctt gtctgctgca gcatcgttct gtgttgtctc tgtctgactg tgtttctgta 1801 tttgtctgaa aatatgggcc cgggccagac tgttaccact cccttaagtt tgaccttagg 1861 tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg 1921 ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg 1981 cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca 2041 tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc 2101 tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc 2161 gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct 2221 cactccttct ctaggcgccc ccatatggcc atatgagatc ttatatgggg cacccccgcc 2281 ccttgtaaac ttccctgacc ctgacatgac aagagttact aacagcccct ctctccaagc 2341 tcacttacag gctctctact tagtccagca cgaagtctgg agacctctgg cggcagccta 2401 ccaagaacaa ctggaccgac cggtggtacc tcacccttac cgagtcggcg acacagtgtg 2461 ggtccgccga caccagacta agaacctaga acctcgctgg aaaggacctt acacagtcct 2521 gctgaccacc cccaccgccc tcaaagtaga cggcatcgca gcttggatac acgccgccca 2581 cgtgaaggct gccgaccccg ggggtggacc atcctctaga ctgccggatc cgccaccatg 2641 ggagtgcagg tggaaaccat ctccccagga gacgggcgca ccttccccaa gcgcggccag 2701 acctgtgtgg tgcactacac cgggatgctt gaagatggaa agaaagtcga ttcctcccgg 2761 gacagaaaca agccctttaa gtttatgcta ggcaagcagg aggtgatccg aggctgggaa 2821 gaaggggttg cccagatgag tgtgggtcag agagccaaac tgactatatc tccagattat 2881 gcctatggtg ccactgggca cccaggcatc atcccaccac atgccactct cgtcttcgat 2941 gtggagcttc taaaaccgga agaattcgtg agcaagggcg aggagctgtt caccggggtg 3001 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc 3061 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc 3121 aagctgcccg tgccctggcc caccctcgtg accaccttcg gctacggcct gcagtgcttc 3181 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc 3241 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag 3301 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag 3361 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat 3421 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc 3481 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc 3541 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgag caaagacccc 3601 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc 3661 ggcatggacg agctgtacaa ggtcgactat ccgtacgacg taccagacta cgcactcgac 3721 taactcgagg gccggcgcgc cgcggccgct acgtaaattc csshcccccc cccctctccc 3781 yccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct 3841 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc 3901 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc 3961 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg 4021 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa 4081 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt 4141 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg 4201 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta 4261 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt 4321 tcctttgaaa aacacgatga taatatggcc acaaccatgg tgagcggcct gctgaaggag 4381 agtatgcgca tcaagatgta catggagggc accgtgaacg gccactactt caagtgcgag 4441 ggcgagggcg acggcaaccc cttcgccggc acccagagca tgagaatcca cgtgaccgag 4501 ggcgcccccc tgcccttcgc cttcgacatc ctggccccct gctgcgagta cggcagcagg 4561 accttcgtgc accacaccgc cgagatcccc gacttcttca agcagagctt ccccgagggc 4621 ttcacctggg agagaaccac cacctacgag gacggcggca tcctgaccgc ccaccaggac 4681 accagcctgg agggcaactg cctgatctac aaggtgaagg tgcacggcac caacttcccc 4741 gccgacggcc ccgtgatgaa gaacaagagc ggcggctggg agcccagcac cgaggtggtg 4801 taccccgaga acggcgtgct gtgcggccgg aacgtgatgg ccctgaaggt gggcgaccgg 4861 cacctgatct gccaccacta caccagctac cggagcaaga aggccgtgcg cgccctgacc 4921 atgcccggct tccacttcac cgacatccgg ctccagatgc tgcggaagaa gaaggacgag 4981 tacttcgagc tgtacgaggc cagcgtggcc cggtacagcg acctgcccga gaaggccaac 5041 agatctcccg ggatggtgag cggcctgctg aaggagagta tgcgcatcaa gatgtacatg 5101 gagggcaccg tgaacggcca ctacttcaag tgcgagggcg agggcgacgg caaccccttc 5161 gccggcaccc agagcatgag aatccacgtg accgagggcg cccccctgcc cttcgccttc 5221 gacatcctgg ccccctgctg cgagtacggc agcaggacct tcgtgcacca caccgccgag 5281 atccccgact tcttcaagca gagcttcccc gagggcttca cctgggagag aaccaccacc 5341 tacgaggacg gcggcatcct gaccgcccac caggacacca gcctggaggg caactgcctg 5401 atctacaagg tgaaggtgca cggcaccaac ttccccgccg acggccccgt gatgaagaac 5461 aagagcggcg gctgggagcc cagcaccgag gtggtgtacc ccgagaacgg cgtgctgtgc 5521 ggccggaacg tgatggccct gaaggtgggc gaccggcacc tgatctgcca ccactacacc 5581 agctaccgga gcaagaaggc cgtgcgcgcc ctgaccatgc ccggcttcca cttcaccgac 5641 atccggctcc agatgctgcg gaagaagaag gacgagtact tcgagctgta cgaggccagc 5701 gtggcccggt acagcgacct gcccgagaag gccaactgag tcgacgataa aataaaagat 5761 tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct 5821 agcttaagta acgccatttt gcaaggcatg gaaaaataca taactgagaa tagagaagtt 5881 cagatcaagg tcaggaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt 5941 aagcagttcc tgccccggct cagggccaag aacagatgga acagctgaat atgggccaaa 6001 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca 6061 gatgcggtcc agccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa 6121 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg 6181 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggggcgc 6241 cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt 6301 tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc 6361 cgtcagcggg ggtctttcac atgcagcatg tatcaaaatt aatttggttt tttttcttaa 6421 gtatttacat taaatggcca tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg 6481 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 6541 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 6601 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 6661 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 6721 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 6781 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 6841 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 6901 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 6961 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 7021 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 7081 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg 7141 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 7201 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 7261 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 7321 cacgttaagg gattttggtc atgagacaat aaccctgata aatgcttcaa taatattgaa 7381 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 7441 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 7501 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 7561 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 7621 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 7681 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 7741 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 7801 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 7861 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 7921 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 7981 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 8041 cacttctgcg ctcggccctt ccggctg //