LOCUS Exported 6275 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION PI(DM): Photo-inhibitable/light sensitive; DPZ: DH-PH-ZnF domains of Vav2. ACCESSION . VERSION . KEYWORDS mCherry-PI(WT)-Vav2(DPZ) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6275) 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 6275) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6275 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(2..21) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 248..268 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter complement(293..397) /gene="bla" /label=AmpR promoter rep_origin 424..879 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(511..530) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 721..742 /label=F1ori-F /note="F1 origin, forward primer" polyA_signal 886..1007 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(897..916) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 951..970 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS complement(2763..3470) /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(3038..3057) /label=mCherry-F /note="mCherry, forward primer" primer_bind 3307..3325 /label=mCherry-R /note="mCherry, reverse primer" promoter complement(3502..3705) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(3531..3551) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 3706..4009 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" rep_origin complement(4184..4772) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4264..4283) /label=pBR322ori-F /note="pBR322 origin, forward primer" polyA_signal 5101..5148 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" primer_bind 5173..5192 /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" CDS complement(5380..6174) /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5491..5510) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 6101..6120 /label=Neo-R /note="Neomycin resistance gene, reverse primer" promoter complement(6209..291) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6223..83 /label=SV40 ori /note="SV40 origin of replication" ORIGIN 1 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa 61 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa 121 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg 181 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac 241 tttccacacc ctaactgaca cacattccac agctggttct ttccgcctca ggactcttcc 301 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 361 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 421 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 481 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg 541 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat 601 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg 661 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 721 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt 781 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat 841 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta cgccttaaga tacattgatg 901 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg 961 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt 1021 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa 1081 acctctacaa atgtggtatg gctgattatg atcagttatc tagattatgc tccaggagca 1141 tccacatctg caggtgaact catcttgcag ggggggatca cctccaggca ttccttgtgt 1201 gccccgacgc cacatctggt acacaggtat ccctggtaga aggtacccct gagaaacatc 1261 ttgcaggctt tgcagttggt agtcttgtcg aatgtgtaca tctggaagct atgatggttg 1321 gcattggcct tatctggctt gatgtttgac atggccatct cgaactgctc catccacttc 1381 cgcttcatgt cttccgtctt gcagaagaac tgaaagcctt gctttccttg gaggtgaatc 1441 aggtagaagc cataggacca cttcttgatg tccttgttgt gcatcgggtc atcggtcatc 1501 ttgtggaaga gcagctcaat gacctccttc agctcatagc tgtagccctt cctcttgcac 1561 acgatgacca ccttgtcaaa taggaacagg tacctgtctt gcttggtgtg gttgactatg 1621 gaccggactt taagctcccc gtcaatcttt ggcctcccaa attcctccag cttcacttgc 1681 aggttttcta tggagcactg gaactcgcta atcttcttca aggtctcctt gtcccgcttc 1741 acttcattga tgtacatggc caagtcctgc atggcttcca gggcttcttt gagctgttgt 1801 ctttctggtc ggtcaagttc ttttgccgcc tcatcaatat tttctgcagt tttcttaatc 1861 agcatgactc cctctctctc ggcagcatct cggacatgct cagttccatc caactgaacc 1921 ccaataaagt actggacatc tcccttctga tctcgcatag gctgcaagtg aaagaggttc 1981 cagaactttt taccactctt tgtataatta atcagctgaa cagtgacctc tgtttggtta 2041 tctatggcat ctctaatttt tctcactgtc gcgcgatcag tttcaggacc ttgtagaaac 2101 ctgcagtttc ttcccaaaat ttcttcacgg ctatattctg tcaactgcaa gaaactatcg 2161 gacgcgaata taatgggatt atctggcaat cttgggtcag taatgacaaa gttcttctca 2221 atacgttcaa gtgtagtagc caatgcagaa tggctcagga gctccttgag cagcaggtgg 2281 tacttcagca cccgttgcat gggcaccacc agcaggtctt gcagcttgaa cttgccatcc 2341 tgaaccctga gtgtgcactc ctccactttc tgcctgaagt cctctcggct ggcgaggagc 2401 tggttcagtg tactctgagc gtgttccatg tggctacagt actctccata gatcaggagc 2461 ctttccttaa actccagaaa gaccttagcc agggtactgc caccagccat catggacaca 2521 tcgatggctc gcagaaagct gtgatgcacc ttgatgaggt cctccaggtt gatgaagaca 2581 gcagccatat ccgccgggct cagcaccagc cgcaagggac ccatgtagtt cttctcaatg 2641 tcctccaggg tgcggtagta cttggcctcg gtctcctgaa tctctaacaa gcagcagctt 2701 ctcttgtcgt cctcagtcat gcccattttc atgggctgct gcacctccac cttgattccg 2761 gacttgtaca gctcgtccat gccgccggtg gagtggcggc cctcggcgcg ttcgtactgt 2821 tccacgatgg tgtagtcctc gttgtgggag gtgatgtcca acttgatgtt gacgttgtag 2881 gcgccgggca gctgcacggg cttcttggcc ttgtaggtgg tcttgacctc agcgtcgtag 2941 tggccgccgt ccttcagctt cagcctctgc ttgatctcgc ccttcagggc gccgtcctcg 3001 gggtacatcc gctcggagga ggcctcccag cccatggtct tcttctgcat tacggggccg 3061 tcggagggga agttggtgcc gcgcagcttc accttgtaga tgaactcgcc gtcctgcagg 3121 gaggagtcct gggtcacggt caccacgccg ccgtcctcga agttcatcac gcgctcccac 3181 ttgaagccct cggggaagga cagcttcaag tagtcgggga tgtcggcggg gtgcttcacg 3241 taggccttgg agccgtacat gaactgaggg gacaggatgt cccaggcgaa gggcaggggg 3301 ccacccttgg tcaccttcag cttggcggtc tgggtgccct cgtaggggcg gccctcgccc 3361 tcgccctcga tctcgaactc gtggccgttc acggagccct ccatgtgcac cttgaagcgc 3421 atgaactcct tgatgatggc catgttatcc tcctcgccct tgctcaccat ggtgctagcg 3481 gatctgacgg ttcactaaac cagctctgct tatatagacc tcccaccgta cacgcctacc 3541 gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg 3601 gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc 3661 aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg taatagcgat 3721 gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata 3781 atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata 3841 cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg 3901 gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg 3961 gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat 4021 atatgggcta tgaactaatg accccgtaat tgattactat taataactaa tgcatggcgg 4081 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 4141 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 4201 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 4261 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 4321 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 4381 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 4441 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 4501 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 4561 cgaggtatgt aggcggtgat acagagttct tgaagtggtg gcctaactac ggctacacta 4621 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 4681 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 4741 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 4801 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 4861 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 4921 atgagtaacc tgaggctatg gcagggcctg ccgccccgac gttggctgcg agccctgggc 4981 cttcacccga acttgggggg tggggtgggg aaaaggaaga aacgcgggcg tattggcccc 5041 aatggggtct cggtggggta tcgacagagt gccagccctg ggaccgaacc ccgcgtttat 5101 gaacaaacga cccaacaccg tgcgttttat tctgtctttt tattgccgtc atagcgcggg 5161 ttccttccgg tattgtctcc ttccgtgttt cagttagcct ccccctaggg tgggcgaaga 5221 actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc 5281 cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg 5341 gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct cagaagaact cgtcaagaag 5401 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg 5461 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg 5521 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc 5581 caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg 5641 catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc 5701 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg 5761 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc 5821 atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc 5881 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc 5941 tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cttgcagttc 6001 attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag 6061 ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag 6121 cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa 6181 cgatcctcat cctgtctctt gatcgatctt tgcaaaagcc taggcctcca aaaaagcctc 6241 ctcactactt ctggaatagc tcagaggccg aggcg //