LOCUS Exported 10093 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION expression of EGFP tagged Cdc42BPB. ACCESSION . VERSION . KEYWORDS pEGFP-N1-Cdc42BPB SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10093) AUTHORS Ando K, Fukuhara S, Moriya T, Obara Y, Nakahata N, Mochizuki N TITLE Rap1 potentiates endothelial cell junctions by spatially controlling myosin II activity and actin organization. JOURNAL J Cell Biol. 2013 Sep 16;202(6):901-16. doi: 10.1083/jcb.201301115. Epub 2013 Sep 9. PUBMED 24019534 REFERENCE 2 (bases 1 to 10093) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10093 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 6033..6042 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 6039..6758 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 6039..6755 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(6084..6105) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(6345..6364) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 6692..6713 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 6881..7002 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6918..6937) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6972..6991 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(7009..7464) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7146..7167) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 7358..7377 /label=F1ori-R /note="F1 origin, reverse primer" promoter 7491..7595 /gene="bla" /label=AmpR promoter promoter 7597..7954 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(7620..7640) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 7805..7940 /label=SV40 ori /note="SV40 origin of replication" primer_bind 7867..7886 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 7989..8783 /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(8043..8062) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 8653..8672 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(8971..8990) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 9015..9062 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 9391..9979 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9880..9899 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggactcag atctcgagct caagcttcga attctgcagt cgacccacgc gtccgcggac 661 gcgtggggag cggtccatgg acccgcgggg cccggcgccc cagactctgc gccgtcggga 721 cggagcccaa gatgtcggcc taggccgggg cgcgacgacg cggacggggc ggcgaggagg 781 cgccgctgct gccggggctc gcagccgccg agcccccgag ggcgcgccct gacggactgg 841 ccgagccggc ggtgagaggc cggcgcgtcg ggagcgggcc gcgcggcacc atgtcggcca 901 aggtgcggct caagaagctg gagcagctgc tcctggacgg gccctggcgc aacgagagcg 961 ccctgagcgt ggaaacgctg ctcgacgtgc tcgtctgcct gtacaccgag tgcagccact 1021 cggccctgcg ccgcgacaag tacgtggccg agttcctcga gtgggctaaa ccatttacac 1081 agctggtgaa agaaatgcag cttcatcgag aagactttga aataattaaa gtaattggaa 1141 gaggtgcttt tggtgaggtt gctgttgtca aaatgaagaa tactgaacga atttatgcaa 1201 tgaaaatcct caacaagtgg gagatgctga aaagagcaga gaccgcgtgc ttccgagagg 1261 agcgcgatgt gctggtgaac ggcgactgcc agtggatcac cgcgctgcac tacgcctttc 1321 aggacgagaa ccacctgtat ttagtcatgg attactatgt gggtggtgat ttactgaccc 1381 tgctcagcaa atttgaagac aagcttccgg aagatatggc gaggttctac attggtgaaa 1441 tggtgctggc cattgactcc atccatcagc ttcattacgt gcacagagac attaaacctg 1501 acaatgtcct tttggacgtg aatggtcata tccgcctggc tgactttgga tcatgtttga 1561 agatgaatga tgatggcact gtgcagtcct ccgtggccgt gggcacacct gactacatct 1621 cgccggagat cctgcaggcg atggaggacg gcatgggcaa atacgggcct gagtgtgact 1681 ggtggtctct gggtgtctgc atgtatgaga tgctctatgg agaaacgccg ttttatgcgg 1741 agtcactcgt ggagacctat gggaagatca tgaaccatga agagcgattc cagttcccat 1801 cccatgtcac ggatgtatct gaagaagcga aggacctcat ccagagactg atctgcagta 1861 gagaacgccg gctggggcag aatggaatag aggatttcaa aaagcatgcg ttttttgaag 1921 gtctaaattg ggaaaatata cgaaacctag aagcacctta tattcctgat gtgagcagtc 1981 cctctgacac atccaacttc gacgtggatg acgacgtgct gagaaacacg gaaatattac 2041 ctcctggttc tcacacaggc ttttctggat tacatttgcc attcattggt tttacattca 2101 caacggaaag ctgtttttct gatcgaggct ctctgaagag cataatgcag tccaacacat 2161 taaccaaaga tgaggatgtg cagcgggacc tggagcacag cctgcagatg gaagcttacg 2221 agaggaggat tcggaggctg gaacaggaga agctggagct gagcaggaag ctgcaagagt 2281 ccacccagac cgtgcagtcc ctccacggct catctcgggc cctcagcaat tcaaaccgag 2341 ataaagaaat caaaaagcta aatgaagaaa tcgaacgctt gaagaataaa atagcagatt 2401 caaacaggct cgagcgacag cttgaggaca cagtggcgct tcgccaagag cgtgaggact 2461 ccacgcagcg gctgcggggg ctggagaagc agcaccgcgt ggtccggcag gagaaggagg 2521 agctgcacaa gcaactggtt gaagcctcag agcggttgaa atcccaggcc aaggaactca 2581 aagatgccca tcagcagcga aagctggccc tgcaggagtt ctcggagctg aacgagcgca 2641 tggcagagct ccgtgcccag aagcagaagg tgtcccggca gctgcgagac aaggaggagg 2701 agatggaggt ggccacgcag aaggtggacg ccatgcggca ggaaatgcgg agagctgaga 2761 agctcaggaa agagctggaa gctcagcttg atgatgctgt tgctgaggcc tccaaggagc 2821 gcaagcttcg tgagcacagc gagaacttct gcaagcaaat ggaaagcgag ctggaggccc 2881 tcaaggtgaa gcaaggaggc cggggagcgg gtgccacctt agagcaccag caagagattt 2941 ccaaaatcaa atccgagctg gagaagaaag tcttatttta tgaagaggaa ttggtcagac 3001 gtgaggcctc ccatgtgcta gaagtgaaaa atgtgaagaa ggaggtgcat gattcagaaa 3061 gccaccagct ggccctgcag aaagaaatct tgatgttaaa agataagtta gaaaagtcaa 3121 agcgagaacg gcataacgag atggaggagg cagtaggtac aataaaagat aaatacgaac 3181 gagaaagagc gatgctgttt gatgaaaaca agaagctaac tgctgaaaat gaaaagctct 3241 gttcctttgt ggataaactc acagctcaaa atagacagct ggaggatgag ctgcaggatc 3301 tggcagccaa gaaggagtca gtggcccact gggaagctca gattgcggaa atcattcagt 3361 gggtcagtga cgagaaagat gcccggggtt accttcaagc tcttgcttcc aagatgaccg 3421 aagagctcga ggctttgagg agttctagtc tggggtcaag aacactggac ccgctgtgga 3481 aggtgcgccg cagccagaag ctggacatgt ccgcgcggct ggagctgcag tcggccctgg 3541 aggcggagat ccgggccaag cagcttgtcc aggaggagct caggaaggtc aaggacgcca 3601 acctcacctt ggaaagcaaa ctaaaggatt ccgaagccaa aaacagagaa ttattagaag 3661 aaatggaaat tttgaagaaa aagatggaag aaaaattcag agcagatact gggctcaaac 3721 ttccagattt tcaggattcc atttttgagt atttcaacac tgctcctctt gcacatgacc 3781 tgacatttag aaccagctca gctagtgagc aagaaacaca agctccgaag ccagaagcgt 3841 ccccgtcgat gtctgtggct gcatcagagc agcaggagga catggctcgg cccccgcaga 3901 ggccatccgc tgtgccgttg cccaccacgc aggccctggc tctggctgga ccgaagccaa 3961 aagctcacca gttcagcatc aagtccttct ccagccctac tcagtgcagc cactgcacct 4021 ccctgatggt tgggctgatc cggcagggct acgcctgcga ggtgtgttcc tttgcttgcc 4081 acgtgtcctg caaagacggt gccccccagg tgtgcccaat acctcccgag cagtccaaga 4141 ggcctctggg cgtggacgtg cagcgaggca tcggaacagc ctacaaaggc catgtcaagg 4201 tcccaaagcc cacgggggtg aagaagggat ggcagcgcgc atatgcagtc gtctgtgact 4261 gcaagctctt cctgtatgat ctgcctgaag gaaaatccac ccagcctggt gtcattgcga 4321 gccaagtctt ggatctcaga gatgacgagt tttccgtgag ctcagtcctg gcctcagatg 4381 tcattcatgc tacacgccga gatattccat gtatattcag ggtgacggcc tctctcttag 4441 gtgcaccttc taagaccagc tcgctgctca ttctgacaga aaatgagaat gaaaagaaga 4501 agtgggttgg gattctagaa ggactccagt ccatccttca taaaaaccgg ctgaggaatc 4561 aggtcgtgca tgttcccttg gaagcctacg acagctcgct gcctctcatc aaggccatcc 4621 tgacagctgc catcgtggat gcagacagga ttgcagtcgg cctagaagaa gggctctatg 4681 tcatagaggt cacccgagat gtgatcgtcc gtgccgctga ctgtaagaag gtacaccaga 4741 tcgagcttgc tcccagggag aagatcgtaa tcctcctctg tggccggaac caccatgtgc 4801 acctctatcc gtggtcgtcc cttgatggag cggaaggcag ctttgacatc aagcttccgg 4861 aaaccaaagg ctgccagctc atggccacgg ccacactcaa gaggaactct ggcacctgcc 4921 tgtttgtggc cgtgaaacgg ctgatccttt gctatgagat ccagagaacg aagccattcc 4981 acagaaagtt caatgagatt gtggctcccg gcagcgtgca gtgcctggcg gtgctcaggg 5041 acaggctctg tgtgggctac ccttctgggt tctgcctgct gagcatccag ggggacgggc 5101 agcctctaaa cctggtaaat cccaatgacc cctcgcttgc gttcctctca caacagtctt 5161 ttgatgccct ttgtgctgtg gagctcgaaa gcgaggagta cctgctttgc ttcagccaca 5221 tgggactgta cgtggacccg caaggccgga gggcacgcgc gcaggagctc atgtggcctg 5281 cggctcctgt cgcctgtagt tgcagcccca cccacgtcac ggtgtacagc gagtatggcg 5341 tggacgtctt tgatgtgcgc accatggagt gggtgcagac catcggcctg cggaggataa 5401 ggcccctgaa ctctgaaggc accctcaacc tcctcaactg cgagcctcca cgcttgatct 5461 acttcaagag caagttctcg ggagcggttc tcaacgtgcc ggacacctcc gacaacagca 5521 agaagcagat gctgcgcacc aggagcaaaa ggcggttcgt cttcaaggtc ccagaggaag 5581 agagactgca gcagaggcga gagatgctta gagacccaga attgagatcc aaaatgatat 5641 ccaacccaac caacttcaac cacgtggccc acatgggccc aggcgacggc atgcaggtgc 5701 tcatggacct gcctctgagt gctgtgcccc cctcccagga ggaaaggccg ggccccgctc 5761 ccaccaacct ggctcgccag cctccatcca ggaacaagcc ctacatctcg tggccctcat 5821 caggtggatc ggagcctagc gtgactgtgc ctctgagaag tatgtctgat ccagaccagg 5881 actttgacaa agagcctgat tcggactcca ccaaacactc aactccatcg aatagctcca 5941 accccagcgg cccaccgagc cccaactccc cccacaggag ccagctcccc ctcgaaggcc 6001 tggagcagcc ggcctgtgac accgcaccgg tcgccaccat ggtgagcaag ggcgaggagc 6061 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt 6121 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca 6181 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg 6241 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg 6301 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca 6361 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg 6421 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca 6481 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga 6541 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc 6601 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc 6661 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg 6721 ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat 6781 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc 6841 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta 6901 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 6961 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg 7021 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat 7081 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg 7141 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc 7201 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt 7261 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag 7321 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg 7381 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc 7441 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc 7501 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct 7561 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg 7621 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 7681 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg 7741 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc 7801 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat 7861 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg 7921 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat 7981 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga 8041 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc 8101 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga 8161 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg 8221 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc 8281 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg 8341 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga 8401 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc 8461 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca 8521 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg 8581 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct 8641 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg 8701 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc 8761 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac 8821 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt 8881 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga 8941 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg 9001 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt 9061 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca 9121 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga 9181 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc 9241 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat 9301 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc 9361 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg 9421 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct 9481 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct 9541 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct 9601 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg 9661 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc 9721 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga 9781 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 9841 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta 9901 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 9961 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 10021 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat 10081 taccgccatg cat //