LOCUS Exported 8679 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION PI(WT): Photo-inhibitable/blue light sensitive ; WT: wild type. ACCESSION . VERSION . KEYWORDS pBabe-PI(WT)-Src(WT)-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8679) 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 8679) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8679 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 137..156 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 291..1316 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" primer_bind complement(1793..1810) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1964..2552) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2044..2063) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2712..3572) /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 3335..3354 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3573..3677) /gene="bla" /label=AmpR promoter LTR 3708..4177 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 4242..4441 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 4642..5058 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 4988..5010 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 5026..5042 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" protein_bind 5099..5369 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" promoter 5402..5440 /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5437..5461 /label=LNCX /note="Human CMV promoter, forward primer" regulatory 5553..5562 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 7605..7614 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 7611..8321 /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(7756..7774) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 8024..8043 /label=mCherry-F /note="mCherry, forward primer" protein_bind 8335..8351 /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)." promoter 8384..204 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(8419..8442) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" ORIGIN 1 ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga 61 aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg 121 tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc 181 tcttcctcat ctccgggcct ttcgacctgc agccaataag cttgccagct ggggcgccct 241 ctggtaaggt tgggaagccc tgcagatctg atcccggggg gcaatgagat atgaaaaagc 301 ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg 361 acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc 421 gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt 481 atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattta 541 gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc 601 ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg 661 cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat 721 acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa 781 ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt 841 gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg 901 tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg 961 attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc 1021 agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg 1081 cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg 1141 atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg 1201 tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag 1261 tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagagta 1321 gatgccgacc gggatctatc gataaaataa aagattttat ttagtctcca gaaaaagggg 1381 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag 1441 gcatggaaaa atacataact gagaatagag aagttcagat caaggtagga gatccctgag 1501 cccacaaccc ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt 1561 gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg 1621 gtctcctctg agtgattgac tacccgtcag cgggggtctt tcacatgcag catgtatcaa 1681 aattaatttg gttttttttc ttaagtattt acattaaatg gccatagttg cattaatgaa 1741 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 1801 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 1861 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 1921 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 1981 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 2041 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 2101 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 2161 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 2221 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 2281 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 2341 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 2401 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 2461 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 2521 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 2581 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 2641 ggatcttcac ctagatcctt ttgcggccgc aaatcaatct aaagtatata tgagtaaact 2701 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 2761 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 2821 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 2881 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 2941 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 3001 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 3061 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 3121 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 3181 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 3241 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 3301 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 3361 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 3421 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 3481 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 3541 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 3601 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 3661 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcagcctg aatatgggcc 3721 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac 3781 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc 3841 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat 3901 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca 3961 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca 4021 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc 4081 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc 4141 tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt ccgggatcgg 4201 gagacccctg cccagggacc accgacccac caccgggagg caagctggcc agcaacttat 4261 ctgtgtctgt ccgattgtct agtgtctatg actgatttta tgcgcctgcg tcggtactag 4321 ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc 4381 ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga 4441 ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag 4501 aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc 4561 gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt 4621 ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt 4681 cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt 4741 tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc 4801 acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat 4861 ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct 4921 ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg 4981 tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc 5041 actccttctc taggcgccgg ccggatccac gaggcccttt cgtcgtcgag tttaccactc 5101 cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa 5161 aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta 5221 ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat 5281 agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg 5341 agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagctcggt acccgggtcg 5401 aggtaggcgt gtacggtggg aggcctatat aagcagagct cgtttagtga accgtcagat 5461 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag 5521 cctccgcggc cccagatccg taaagtgaat tcgccaccat gggcagcaac aagagcaagc 5581 ccaaggacgc cagccagcgg cgccgcagcc tggagccctc ggaaaacgtg cacggggcag 5641 ggggcgcctt cccggcctca cagacaccga gcaagcccgc ctccgccgac ggccaccgcg 5701 ggcccagcgc cgccttcgtg ccgcccgcgg ccgagcccaa gctcttcgga ggcttcaact 5761 cctcggacac cgtcacctcc ccgcagaggg cggggcctct ggcaggtggg gtgaccacct 5821 ttgtggccct ctatgactat gagtcacgga cagagactga cctgtccttc aagaaagggg 5881 agcggctgca gattgttaac aacacggagg gagactggtg gctggcacac tcgctgagca 5941 cgggacagac cggttacatc cccagcaact atgtggcgcc ctccgactcc atccaggctg 6001 aggagtggta ctttggcaag atcactagac gggaatcaga gcggctgctg ctcaacgccg 6061 agaacccgag agggaccttc ctcgtgaggg agagtgagac cacaaaaggt gcctactgcc 6121 tctctgtatc cgacttcgac aatgccaagg gcctaaatgt gaaacactac aagatccgca 6181 agctggacag cggcggtttc tacatcacct cccgcaccca gttcaacagc ctgcagcagc 6241 tcgtggctta ctactccaaa catgctgatg gcctgtgtca ccgcctcact accgtatgtc 6301 ccacatccaa gcctcagacc cagggattgg ccaaggatgc gtgggagatc ccccgggagt 6361 ccctgcggct ggaggtcaag ctgggccagg gttgcttcgg agaggtgtgg atggggacct 6421 ggaacttggc tactacactt gaacgtattg agaagaactt tgtcattact gacccaagat 6481 tgccagataa tcccattata ttcgcgtccg atagtttctt gcagttgaca gaatatagcc 6541 gtgaagaaat tttgggaaga aactgcaggt ttctacaagg tcctgaaact gatcgcgcga 6601 cagtgagaaa aattagagat gccatagata accaaacaga ggtcactgtt cagctgatta 6661 attatacaaa gagtggtaaa aagttctgga acctctttca cttgcagcct atgcgagatc 6721 agaagggaga tgtccagtac tttattgggg ttcagttgga tggaactgag catgtccgag 6781 atgctgccga gagagaggga gtcatgctga ttaagaaaac tgcagaaaat attgatgagg 6841 cggcaaaaga acttaccacg agggttgcca tcaaaactct gaagccaggc accatgtccc 6901 cagaggcctt cctgcaggag gcccaagtca tgaagaaact gaggcacgag aaactggtgc 6961 agctgtatgc tgtggtgtcg gaagaaccca tttacattgt gacagagtac atgaacaagg 7021 ggagtctgct ggactttctc aagggggaaa cgggcaaata tttgcggcta ccccagctgg 7081 tggacatgtc tgctcagatc gcttcaggca tggcctatgt ggagcggatg aactatgtgc 7141 accgggacct tcgagccgcc aatatcctag taggggagaa cctggtgtgc aaagtggccg 7201 actttgggtt ggcccggctc atagaagaca acgaatacac agcccggcaa ggtgccaaat 7261 tccccatcaa gtggaccgcc cctgaagctg ctctgtacgg caggttcacc atcaagtcgg 7321 atgtgtggtc ctttgggatt ctgctgaccg agctcaccac taagggaaga gtgccctatc 7381 ctgggatggt gaaccgtgag gttctggacc aggtggagcg gggctaccgg atgccttgtc 7441 cccccgagtg ccccgagtcc ctgcatgacc ttatgtgcca gtgctggcgg aaggagcccg 7501 aggagcggcc caccttcgag tacctgcagg ccttcctgga agactacttt acgtccactg 7561 agccacagta ccagcccggg gagaacctat cggatccacc ggtcgccacc atggtgagca 7621 agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg 7681 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg 7741 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg 7801 acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca 7861 tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact 7921 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca 7981 tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga 8041 agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg 8101 gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga 8161 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt 8221 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg 8281 gccgccactc caccggcggc atggacgagc tgtacaagta agtcgagtgt ggaattgtga 8341 gcggataaca atttgtcgag ggcccctgca ggtcaattct accgggtagg ggaggcgctt 8401 ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca 8461 agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc 8521 tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc 8581 ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg 8641 cagatggaca gcaccgctga gcaatggaag cgggtaggc //