LOCUS Exported 5269 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of human Rap2b. ACCESSION . VERSION . KEYWORDS mCherry-Rap2b SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5269) AUTHORS Liu C, Takahashi M, Li Y, Dillon TJ, Kaech S, Stork PJ TITLE The interaction of Epac1 and Ran promotes Rap1 activation at the nuclear envelope. JOURNAL Mol Cell Biol. 2010 Aug;30(16):3956-69. doi: 10.1128/MCB.00242-10. Epub 2010 Jun 14. PUBMED 20547757 REFERENCE 2 (bases 1 to 5269) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5269 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 113..416 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 417..620 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 571..591 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 659..668 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 665..1372 /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(810..828) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1078..1097 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 2109..2230 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2146..2165) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2200..2219 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2237..2692) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2374..2395) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2586..2605 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2719..2823 /gene="bla" /label=AmpR promoter promoter 2825..3182 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2848..2868) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3033..3168 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3095..3114 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3217..4011 /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(3271..3290) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3881..3900 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4199..4218) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4243..4290 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4619..5207 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5108..5127 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt 61 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat 121 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa 181 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg 241 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc 301 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct 361 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga 421 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa 481 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc 541 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg 601 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggtcgc 661 caccatggtg agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt 721 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga 781 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct 841 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa 901 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga 961 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca 1021 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc 1081 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga 1141 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga 1201 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa 1261 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta 1321 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtccggact 1381 cagatctcga gctcaagctt cgaattctat gagagagtac aaagtggtgg tgctgggctc 1441 gggcggcgtg ggcaagtccg cgctcaccgt gcagttcgtg acgggctcct tcatcgagaa 1501 gtacgacccg accatcgaag acttttaccg caaggagatt gaggtggact cgtcgccgtc 1561 ggtgctggag atcctggata cggcgggcac cgagcagttc gcgtccatgc gggacctgta 1621 catcaagaac ggccagggct tcatcctggt ctacagcctc gtcaaccagc agagcttcca 1681 ggacatcaag cccatgcggg accagatcat ccgcgtgaag cggtacgagc gcgtgcccat 1741 gatcctggtg ggcaacaagg tggacctgga gggtgagcgc gaggtctcgt acggggaggg 1801 caaggccctg gctgaggagt ggagctgccc cttcatggag acgtcggcca aaaacaaagc 1861 ctcggtagac gagctatttg ccgagatcgt gcggcagatg aactacgcgg cgcagcccaa 1921 cggcgatgag ggctgctgct cggcctgcgt gatcctctga ctcgagtcta gagggcccgg 1981 gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac 2041 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg 2101 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 2161 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 2221 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt 2281 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca 2341 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta 2401 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta 2461 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg 2521 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga 2581 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg 2641 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc 2701 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat 2761 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag 2821 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc 2881 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt 2941 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 3001 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 3061 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 3121 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca 3181 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac 3241 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca 3301 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt 3361 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg 3421 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga 3481 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct 3541 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg 3601 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg 3661 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc 3721 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat 3781 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac 3841 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt 3901 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct 3961 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc 4021 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca 4081 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga 4141 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg 4201 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga 4261 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc 4321 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt 4381 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg 4441 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt 4501 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt 4561 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt 4621 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag 4681 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca 4741 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca 4801 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg 4861 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg 4921 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct 4981 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga 5041 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 5101 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg 5161 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 5221 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt //