LOCUS Exported 6640 bp ds-DNA circular SYN 23-AUG-2021 DEFINITION Expresses Cdc42 Flp-dependently in mammalian cells. ACCESSION . VERSION . KEYWORDS pCAFNF-Cdc42 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6640) AUTHORS Aihara S, Fujimoto S, Sakaguchi R, Imai T TITLE BMPR-2 gates activity-dependent stabilization of primary dendrites during mitral cell remodeling. JOURNAL Cell Rep. 2021 Jun 22;35(12):109276. doi: 10.1016/j.celrep.2021.109276. PUBMED 34161760 REFERENCE 2 (bases 1 to 6640) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 23, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6640 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 5..384 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 663..1679 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1603..1625 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1687..1706 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" protein_bind 1754..1787 /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" CDS 1830..2624 /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(1884..1903) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 2494..2513 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 2804..2938 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" protein_bind 2947..2980 /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" primer_bind complement(3683..3702) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 3748..3803 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(3802..3821) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4164..4180) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4164..4180) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4177..4199) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4188..4204 /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)." promoter complement(4212..4242) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4257..4278 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin 4363..4497 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4424..4443 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 4517..4651 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4719..4736) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4890..5478) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4970..4989) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5649..6509) /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 6272..6291 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6510..6614) /gene="bla" /label=AmpR promoter ORIGIN 1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat 61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg 121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata 181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta 241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc 301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc 421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca 481 gcgatggggg cggggggggg gggggnsgcg cgccrggsgg ggsggggsgg ggsgrggggs 541 ggggsggggs gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt 601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg 661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc 721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc 781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg 841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt 901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc 961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg 1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg 1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc 1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc 1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg 1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg 1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg 1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca 1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg 1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc 1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg 1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc 1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgattaatt 1741 cgagcgaacg cgtgaagttc ctattctcta gaaagtatag gaacttccgg atttgatctg 1801 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt 1861 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct 1921 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga 1981 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg 2041 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact 2101 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg 2161 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct 2221 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg 2281 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt 2341 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg 2401 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc 2461 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag 2521 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt 2581 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt 2641 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc 2701 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca 2761 gcgcggggat ctcatgctgg agttcttcgc ccaccccatc gataacttgt ttattgcagc 2821 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 2881 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa 2941 atccgagaag ttcctattct ctagaaagta taggaacttc atctcgagtc gctcggtacg 3001 atttaaattg aattcgccac catgcagaca attaagtgtg ttgttgttgg tgatggtgct 3061 gttggtaaaa catgtctcct gatatcctac acaacaaaca aattcccatc ggaatatgta 3121 ccaactgttt ttgacaacta tgcagtcaca gttatgattg gtggagagcc atacactctt 3181 ggactttttg atactgcagg gcaagaggat tatgacagac tacgaccgct aagttatcca 3241 cagacagatg tttttctagt atgtttctca gtggtctctc catcctcatt tgaaaatgtg 3301 aaagaaaagt gggtgcctga gataactcac cactgtccaa agactccttt cttgcttgtt 3361 gggacccaaa ttgatctcag agatgacccc tctactattg agaaacttgc caagaacaaa 3421 cagaagccta ttactccaga gactgctgaa aagctggcgc gggatctgaa ggctgtcaag 3481 tatgtggagt gctctgccct cacacagaga ggtctgaaga atgtgtttga tgaggctatc 3541 ctagctgccc tcgagcctcc ggaaactcaa cccaaaagga agtgctgtat attctaagcg 3601 gccgcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc 3661 tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga 3721 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt 3781 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac 3841 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga 3901 acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct 3961 tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat 4021 ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc 4081 tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc 4141 agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 4201 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 4261 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 4321 tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 4381 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg 4441 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag 4501 gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag 4561 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 4621 actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg 4681 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 4741 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 4801 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 4861 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 4921 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 4981 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 5041 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 5101 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 5161 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 5221 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 5281 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 5341 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 5401 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 5461 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 5521 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 5581 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 5641 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 5701 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 5761 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 5821 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 5881 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 5941 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 6001 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 6061 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 6121 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 6181 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 6241 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 6301 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 6361 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 6421 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 6481 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 6541 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 6601 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg //