LOCUS Exported 5268 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pRK5-myc-Rac1-wt SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5268) TITLE Bokoch Lab GTPases REFERENCE 2 (bases 1 to 5268) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5268 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 98..115 /label=L4440 /note="L4440 vector, forward primer" protein_bind 232..253 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 268..298 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 306..322 /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)." primer_bind 311..333 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 330..346 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 330..346 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" enhancer 375..754 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 755..958 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 909..929 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 955..979 /label=LNCX /note="Human CMV promoter, forward primer" promoter 1189..1207 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1189..1206 /label=SP6 /note="SP6 promoter, forward primer" primer_bind 1220..1239 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 1286..1295 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1295..1324 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1331..1909 /codon_start=1 /product="human Rac family small GTPase 1" /label=Rac1 /translation="MQAIKCVVVGDGAVGKTCLLISYTTNAFPGEYIPTVFDNYSANVM VDGKPVNLGLWDTAGQEDYDRLRPLSYPQTDVFLICFSLVSPASFENVRAKWYPEVRHH CPNTPIILVGTKLDLRDDKDTIEKLKEKKLTPITYPQGLAMAKEIGAVKYLECSALTQR GLKTVFDEAIRAVLCPPPVKKRKRKCLLL" regulatory 1929..1938 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" polyA_signal 1942..2076 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1979..1998) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2033..2052 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" promoter 2145..2502 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2168..2188) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2353..2488 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2415..2434 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(2522..2539) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2522..2538) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2531..2553) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2751..3206 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2838..2857) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3048..3069 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3223..3242) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3342..3364 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3402..3420) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3488..3592 /gene="bla" /label=AmpR promoter CDS 3593..4453 /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(3811..3830) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4624..5212 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5113..5132 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg 61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc 121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag 181 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag 241 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg 301 tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga ttacgaatta 361 attcgagctc gcccgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc 421 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc 481 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt 541 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca 601 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg 661 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca 721 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa 781 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa 841 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc 901 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg 961 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag 1021 acaccgggac cgatccagcc tccgcggccg ggaacggtgc attggaacgc ggattccccg 1081 tgccaagagt gacgtaagta ccgcctatag agtctatagg cccaccccct tggcttcgtt 1141 agaacgcggc tacaattaat acataacctt atgtatcata cacatacgat ttaggtgaca 1201 ctatagaata acatccactt tgcctttctc tccacaggtg tccactccca ggtccaactg 1261 cacctcggtt ctatcgattg aattggccac catggagcag aagctgatct ccgaggagga 1321 cctgggatcc atgcaggcca tcaagtgtgt ggtggtggga gacggagctg taggtaaaac 1381 ttgcctactg atcagttaca caaccaatgc atttcctgga gaatatatcc ctactgtctt 1441 tgacaattat tctgccaatg ttatggtaga tggaaaaccg gtgaatctgg gcttatggga 1501 tacagctgga caagaagatt atgacagatt acgcccccta tcctatccgc aaacagatgt 1561 gttcttaatt tgcttttccc ttgtgagtcc tgcatcattt gaaaatgtcc gtgcaaagtg 1621 gtatcctgag gtgcggcacc actgtcccaa cactcccatc atcctagtgg gaactaaact 1681 tgatcttagg gatgataaag acacgatcga gaaactgaag gagaagaagc tgactcccat 1741 cacctatccg cagggtctag ccatggctaa ggagattggt gctgtaaaat acctggagtg 1801 ctcggcgctc acacagcgag gcctcaagac agtgtttgac gaagcgatcc gagcagtcct 1861 ctgcccgcct cccgtgaaga agaggaagag aaaatgcctg ctgttgtaag aattcctgca 1921 gaagcttggc cgccatggcc caacttgttt attgcagctt ataatggtta caaataaagc 1981 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 2041 tccaaactca tcaatgtatc ttatcatgtc tggatcggga attaattcgg cgcagcacca 2101 tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac 2161 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga 2221 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc 2281 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc 2341 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 2401 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 2461 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctgtt aacagcttgg 2521 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc 2581 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc 2641 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc 2701 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg 2761 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc 2821 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg 2881 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg 2941 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg 3001 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt 3061 ccaaactgga acaacactca accctatctc gggctattct tttgatttat aagggatttt 3121 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt 3181 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc 3241 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg 3301 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca 3361 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt 3421 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg 3481 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct 3541 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat 3601 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc 3661 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg 3721 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg 3781 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga 3841 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta 3901 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc 3961 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc 4021 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg 4081 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc 4141 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca 4201 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct 4261 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat 4321 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg 4381 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat 4441 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact 4501 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat 4561 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc 4621 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct 4681 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg 4741 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca 4801 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc 4861 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga 4921 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac 4981 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga 5041 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 5101 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg 5161 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag 5221 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctca //