LOCUS Exported 6674 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3-EGFP-Cdc42(Q61L) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6674) AUTHORS Nalbant P, Hodgson L, Kraynov V, Toutchkine A, Hahn KM TITLE Activation of endogenous Cdc42 visualized in living cells. JOURNAL Science. 2004 Sep 10. 305(5690):1615-9. PUBMED 15361624 REFERENCE 2 (bases 1 to 6674) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6674 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 107..128 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 143..173 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 181..197 /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 186..208 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 205..221 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 205..221 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" polyA_signal 270..391 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(281..300) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 335..354 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS complement(565..1359) /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(676..695) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 1286..1305 /label=Neo-R /note="Neomycin resistance gene, reverse primer" promoter complement(1426..1755) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 1440..1575 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(1494..1513) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 1740..1760 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin complement(1769..2197) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1879..1900) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2091..2110 /label=F1ori-R /note="F1 origin, reverse primer" polyA_signal 2243..2467 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind 2456..2473 /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" promoter 2493..2511 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 2493..2510 /label=SP6 /note="SP6 promoter, forward primer" CDS complement(2539..3114) /codon_start=1 /product="constitutively active mutant of human Rho family small GTPase Cdc42" /label=Cdc42 (Q61L) /translation="MQTIKCVVVGDGAVGKTCLLISYTTNKFPSEYVPTVFDNYAVTVM IGGEPYTLGLFDTAGLEDYDRLRPLSYPQTDVFLVCFSVVSPSSFENVKEKWVPEITHH CPKTPFLLVGTQIDLRDDPSTIEKLAKNKQKPITPETAEKLARDLKAVKYVECSALTQK GLKNVFDEAILAALEPPEPKKSRRCVLL" CDS complement(3121..3837) /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS complement(3121..3837) /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3163..3184) /label=EGFP-C /note="EGFP, forward primer" primer_bind 3512..3531 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3771..3792 /label=EGFP-N /note="EGFP, reverse primer" regulatory 3834..3843 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(3856..3875) /label=T7 /note="T7 promoter, forward primer" promoter complement(3857..3875) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter complement(3920..4123) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(3949..3969) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 4124..4503 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" primer_bind 4675..4694 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 4769..4873 /gene="bla" /label=AmpR promoter CDS 4874..5734 /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(5092..5111) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5905..6493 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6394..6413 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6647..6664 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc 121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa 181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct 241 agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac 301 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt 361 tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat 421 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt 481 catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg 541 tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat 601 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc 661 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac 721 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg 781 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag 841 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc 901 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc 961 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga 1021 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa 1081 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc 1141 cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga 1201 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc 1261 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc 1321 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt 1381 ctcttgatca gatccgaaaa tggatataca agctcccggg agctttttgc aaaagcctag 1441 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggcagagg cggcctcggc 1501 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg 1561 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg 1621 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga 1681 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca 1741 ccctaactga cacacattcc acagaattaa ttcgcgttaa atttttgtta aatcagctca 1801 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag 1861 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc 1921 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc 1981 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc 2041 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa 2101 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc 2161 acacccgccg cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag 2221 ctggttcttt ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg 2281 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac 2341 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca 2401 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa 2461 ggcacagtcg aggctgatca gcgagctcta gcatttaggt gacactatag aatagggccc 2521 tctagatgca tgctcgagtc atagcagcac acacctgcgg ctcttcttcg gttctggagg 2581 ctccagggca gccaatattg cttcgtcaaa tacattcttt aggcctttct gtgtaagtgc 2641 agaacactcc acatacttga cagccttcag gtcacgggcc agcttttcag cagtctctgg 2701 agtgataggc ttctgtttgt tcttggcaag tttctcaata gtagaggggt catctctgag 2761 atcaatttga gtcccaacaa gcaagaaagg agtctttgga cagtggtgag ttatctcagg 2821 cacccacttt tctttcacgt tttcaaatga agatggagag accactgaaa aacagactag 2881 aaatacatct gtttgtggat aactcagcgg tcgtaatctg tcataatcct ctagccctgc 2941 agtatcaaaa agtccaagag tatatggttc tccaccaatc ataactgtga ctgcatagtt 3001 gtcaaaaaca gtcggtacat attccgatgg aaatttgttt gttgtgtagg atatcaggag 3061 acatgtttta ccaacagcac catcgcccac aacaacacac ttaattgtct gcatgaattc 3121 cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac 3181 catgtgatcg cgcttctcgt tggggtcttt gctcagggcg gactgggtgc tcaggtagtg 3241 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc 3301 gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg aagttcacct tgatgccgtt 3361 cttctgcttg tcggccatga tatagacgtt gtggctgttg tagttgtact ccagcttgtg 3421 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag 3481 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa 3541 gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt 3601 catgtggtcg gggtagcggc tgaagcactg cacgccgtag gtcagggtgg tcacgagggt 3661 gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta 3721 ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc 3781 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatggt 3841 ggcaagcttg ggtctcccta tagtgagtcg tattaatttc gataagccag taagcagtgg 3901 gttctctagt tagccagaga gctctgctta tatagacctc ccaccgtaca cgcctaccgc 3961 ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt 4021 gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa 4081 accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga 4141 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat 4201 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca 4261 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga 4321 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt 4381 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat 4441 atgggctatg aactaatgac cccgtaattg attactatta ataactagtc aataatcaat 4501 gtcaacgcgt atatctggcc cgtacatcgc gaagcagcgc aaaacgccta accctaagca 4561 gattcttcat gcaattgtcg gtcaagcctt gccttgttgt agcttaaatt ttgctcgcgc 4621 actactcagc gacctccaac acacaagcag ggagcagata ctggcttaac tatgcggcat 4681 cagagcagat tgtactgaga gtgcaccata ggggatcggg agatctcccg atccgtcgac 4741 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 4801 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 4861 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc 4921 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga 4981 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga 5041 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg 5101 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc 5161 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac 5221 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact 5281 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca 5341 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg 5401 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact 5461 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg 5521 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg 5581 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat 5641 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc 5701 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat 5761 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt 5821 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc 5881 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt 5941 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac 6001 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt 6061 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 6121 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 6181 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 6241 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 6301 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt 6361 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc 6421 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg 6481 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc 6541 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc 6601 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag 6661 cgaggaagcg gaag //