LOCUS Exported 7208 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION PI(WT): Photo-inhibitable/blue light sensitive Rac1. ACCESSION . VERSION . KEYWORDS pBabe-PI(WT)-Rac1(WT)-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7208) AUTHORS Dagliyan O, Tarnawski M, Chu PH, Shirvanyants D, Schlichting I, Dokholyan NV, Hahn KM TITLE Engineering extrinsic disorder to control protein activity in living cells. JOURNAL Science. 2016 Dec 16;354(6318):1441-1444. PUBMED 27980211 REFERENCE 2 (bases 1 to 7208) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7208 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 315..338 /label=MSCV-rev /note="Murine stem cell virus, reverse primer" protein_bind 406..422 /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)." CDS complement(1489..2196) /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(1764..1783) /label=mCherry-F /note="mCherry, forward primer" primer_bind 2033..2051 /label=mCherry-R /note="mCherry, reverse primer" regulatory 2193..2202 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(2297..2321) /label=LNCX /note="Human CMV promoter, forward primer" promoter complement(2318..2356) /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" protein_bind 2389..2659 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" CDS complement(2700..3116) /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind complement(2716..2732) /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" primer_bind complement(2748..2770) /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" misc_feature 3317..3516 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" LTR 3581..4050 /note="long terminal repeat from Moloney murine leukemia virus" promoter 4081..4185 /gene="bla" /label=AmpR promoter CDS 4186..5046 /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(4404..4423) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5206..5794 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5695..5714 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5948..5965 /label=L4440 /note="L4440 vector, forward primer" CDS complement(6474..7073) /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(6557..6577) /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind 7054..7073 /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind complement(7144..7163) /label=mPGK-F /note="Mouse PGK promoter, forward primer" ORIGIN 1 cccccggacc caccccttcc cagcctctga gcccagaaag cgaaggagca aagctgctat 61 tggccgctgc cccaaaggcc tacccgcttc cattgctcag cggtgctgtc catctgcacg 121 agactagtga gacgtgctac ttccatttgt cacgtcctgc acgacgcgag ctgcggggcg 181 ggggggaact tcctgactag gggaggagta gaaggtggcg cgaaggggcc accaaagaac 241 ggagccggtt ggcgcctacc ggtggatgtg gaatgtgtgc gaggccagag gccacttgtg 301 tagcgccaag tgcccagcgg ggctgctaaa gcgcatgctc cagactgcct tgggaaaagc 361 gcctccccta cccggtagaa ttgacctgca ggggccctcg acaaattgtt atccgctcac 421 aattccacac tcgagttaca acagcaggca ttttctcttc ctcttcttca cgggaggcgg 481 gcagaggact gctcggatcg cttcgtcaaa cactgtcttg aggcctcgct gtgtgagcgc 541 cgagcactcc aggtatttta cggcaccaat ctccttagcc atggctagac cctgcggata 601 ggtgatggga gtcagcttct tctccttcag tttctcgatc gtgtctttat catccctaag 661 atcaagttta gttcccacta ggatgatggg agtgttggga cagtggtgcc gcacctcagg 721 ataccacttt gcacggacat tttcaaatga tgcaggactc acaagggaaa agcaaattaa 781 gaacacatct gtttgcggat aggatagggg gcgtaatctg tcataatctt cttgtccagc 841 tgtatcccat aagcccagat tcaccggttt tccaccactg ccaagttctt ttgccgcctc 901 atcaatattt tctgcagttt tcttaatcag catgactccc tctctctcgg cagcatctcg 961 gacatgctca gttccatcca actgaacccc aataaagtac tggacatctc ccttctgatc 1021 tcgcataggc tgcaagtgaa agaggttcca gaacttttta ccactctttg tataattaat 1081 cagctgaaca gtgacctctg tttggttatc tatggcatct ctaatttttc tcactgtcgc 1141 gcgatcagtt tcaggacctt gtagaaacct gcagtttctt cccaaaattt cttcacggct 1201 atattctgtc aactgcaaga aactatcgga cgcgaatata atgggattat ctggcaatct 1261 tgggtcagta atgacaaagt tcttctcaat acgttcaagt gtagtagcca aacctgatcc 1321 atctaccata acattggcag aataattgtc aaagacagta gggatatatt ctccaggaaa 1381 tgcattggtt gtgtaactga tcagtaggca agttttacct acagctccgt ctcccaccac 1441 cacacacttg atggcctgca tggatccgag ctcagatctg agtccggact tgtacagctc 1501 gtccatgccg ccggtggagt ggcggccctc ggcgcgttcg tactgttcca cgatggtgta 1561 gtcctcgttg tgggaggtga tgtccaactt gatgttgacg ttgtaggcgc cgggcagctg 1621 cacgggcttc ttggccttgt aggtggtctt gacctcagcg tcgtagtggc cgccgtcctt 1681 cagcttcagc ctctgcttga tctcgccctt cagggcgccg tcctcggggt acatccgctc 1741 ggaggaggcc tcccagccca tggtcttctt ctgcattacg gggccgtcgg aggggaagtt 1801 ggtgccgcgc agcttcacct tgtagatgaa ctcgccgtcc tgcagggagg agtcctgggt 1861 cacggtcacc acgccgccgt cctcgaagtt catcacgcgc tcccacttga agccctcggg 1921 gaaggacagc ttcaagtagt cggggatgtc ggcggggtgc ttcacgtagg ccttggagcc 1981 gtacatgaac tgaggggaca ggatgtccca ggcgaagggc agggggccac ccttggtcac 2041 cttcagcttg gcggtctggg tgccctcgta ggggcggccc tcgccctcgc cctcgatctc 2101 gaactcgtgg ccgttcacgg agccctccat gtgcaccttg aagcgcatga actccttgat 2161 gatggccatg ttatcctcct cgcccttgct caccatggtg gcggcgaatt cactttacgg 2221 atctggggcc gcggaggctg gatcggtccc ggtgtcttct atggaggtca aaacagcgtg 2281 gatggcgtct ccaggcgatc tgacggttca ctaaacgagc tctgcttata taggcctccc 2341 accgtacacg cctacctcga cccgggtacc gagctcgact ttcacttttc tctatcactg 2401 atagggagtg gtaaactcga ctttcacttt tctctatcac tgatagggag tggtaaactc 2461 gactttcact tttctctatc actgataggg agtggtaaac tcgactttca cttttctcta 2521 tcactgatag ggagtggtaa actcgacttt cacttttctc tatcactgat agggagtggt 2581 aaactcgact ttcacttttc tctatcactg atagggagtg gtaaactcga ctttcacttt 2641 tctctatcac tgatagggag tggtaaactc gacgacgaaa gggcctcgtg gatccggccg 2701 gcgcctagag aaggagtgag ggctggataa agggaggatc gaggcggggt cgaacgagga 2761 ggttcaaggg ggagagacgg ggcggatgga ggaagaggag gcggaggctt agggtgtaca 2821 aagggcttga cccagggagg ggggtcaaaa gccaaggctt cccaggtcac gatgtagggg 2881 acctggtctg ggtgtccatg cgggccaggt gaaaagacct tgatcttaac ctgggtgatg 2941 aggtctcggt taaaggtgcc gtctcgcggc catccgacgt taaaggttgg ccattctgca 3001 gagcagaagg taacccaacg tctcttcttg acatctaccg actggttgtg agcgatccgc 3061 tcgacatctt tccagtgacc taaggtcaaa cttaagggag tggtaacagt ctggccctaa 3121 ttttcagaca aatacagaaa cacagtcaga cagagacaac acagaacgat gctgcagcgc 3181 tgcagcagac aagacgcgcg gcttcggttc caaaccgaaa gcaaaaattc agacggaggc 3241 gggaactgtt ttaggttctc gtctcctacc agaaccacat atcctgacgg ggtcggattc 3301 cacatcgact cccttcctca ggtcgggcca caaaaacggc ccccaaagtc cctgggacgt 3361 ctcccagggt tgcggccggg tgttcagaac tcgtcagttc caccacgggt ccgccagata 3421 cagagctagt tagctaacta gtaccgacgc aggcgcataa aatcagtcat agacactaga 3481 caatcggaca gacacagata agttgctggc cagcttgcct cccggtggtg ggtcggtggt 3541 ccctgggcag gggtctcccg atcccggacg agcccccaaa tgaaagaccc ccgctgacgg 3601 gtagtcaatc actcagagga gaccctccca aggaacagcg agaccacaag tcggatgcaa 3661 ctgcaagagg gtttattgga tacacgggta cccgggcgac tcagtcaatc ggaggactgg 3721 cgccccgagt gaggggttgt gggctctttt attgagctcg gggagcagaa gcgcgcgaac 3781 agaagcgaga agcgaactga ttggttagtt caaataaggc acagggtcat ttcaggtcct 3841 tggggcaccc tggaaacatc tgatggttct ctagaaactg ctgagggctg gaccgcatct 3901 ggggaccatc tgttcttggc cctgagccgg ggcaggaact gcttaccaca gatatcctgt 3961 ttggcccata ttcagctgtt ccatctgttc ttggccctga gccggggcag gaactgctta 4021 ccacagatat cctgtttggc ccatattcag gctgcaggtg gcacttttcg gggaaatgtg 4081 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga 4141 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat 4201 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca 4261 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc 4321 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca 4381 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg 4441 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca 4501 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata 4561 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag 4621 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg 4681 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca 4741 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta 4801 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct 4861 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca 4921 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag 4981 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat 5041 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttgcg gccgcaaaag 5101 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc 5161 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt 5221 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt 5281 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat 5341 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc 5401 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa 5461 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg 5521 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag 5581 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag 5641 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa 5701 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt 5761 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg 5821 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc 5881 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac 5941 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct 6001 ccccgcgcgt tggccgattc attaatgcaa ctatggccat ttaatgtaaa tacttaagaa 6061 aaaaaaccaa attaattttg atacatgctg catgtgaaag acccccgctg acgggtagtc 6121 aatcactcag aggagaccct cccaaggaac agcgagacca caagtcggat gcaactgcaa 6181 gagggtttat tggatacacg ggtacccggg cgactcagtc aatcggagga ctggcgcccc 6241 gagtgagggg ttgtgggctc agggatctcc taccttgatc tgaacttctc tattctcagt 6301 tatgtatttt tccatgcctt gcaaaatggc gttacttaag ctagcttgcc aaacctacag 6361 gtggggtctt tcattccccc ctttttctgg agactaaata aaatctttta ttttatcgat 6421 gcatggggtc gtgcgctcct ttcggtcggg cgctgcgggt cgtggggcgg gcgtcaggca 6481 ccgggcttgc gggtcatgca ccaggtgcgc ggtccttcgg gcacctcgac gtcggcggtg 6541 acggtgaagc cgagccgctc gtagaagggg aggttgcggg gcgcggaggt ctccaggaag 6601 gcgggcaccc cggcgcgctc ggccgcctcc actccgggga gcacgacggc gctgcccaga 6661 cccttgccct ggtggtcggg cgagacgccg acggtggcca ggaaccacgc gggctccttg 6721 ggccggtgcg gcgccaggag gccttccatc tgttgctgcg cggccagccg ggaaccgctc 6781 aactcggcca tgcgcgggcc gatctcggcg aacaccgccc ccgcttcgac gctctccggc 6841 gtggtccaga ccgccaccgc ggcgccgtcg tccgcgaccc acaccttgcc gatgtcgagc 6901 ccgacgcgcg tgaggaagag ttcttgcagc tcggtgaccc gctcgatgtg gcggtccgga 6961 tcgacggtgt ggcgcgtggc ggggtagtcg gcgaacgcgg cggcgagggt gcgtacggcc 7021 ctggggacgt cgtcgcgggt ggcgaggcgc accgtgggct tgtactcggt catggtaagc 7081 ttattggctg caggtcgaaa ggcccggaga tgaggaagag gagaacagcg cggcagacgt 7141 gcgcttttga agcgtgcaga atgccgggcc tccggaggac cttcgggcgc ccgccccgcc 7201 cctgagcc //