LOCUS Exported 6474 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION mammalian expression of human SEPT9 with an mCherry fusion. ACCESSION . VERSION . KEYWORDS pmCherry-C1_SEPT9 i1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6474) AUTHORS Dolat L, Hunyara JL, Bowen JR, Karasmanis EP, Elgawly M, Galkin VE, Spiliotis ET TITLE Septins promote stress fiber-mediated maturation of focal adhesions and renal epithelial motility. JOURNAL J Cell Biol. 2014 Oct 27;207(2):225-35. doi: 10.1083/jcb.201405050. PUBMED 25349260 REFERENCE 2 (bases 1 to 6474) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6474 /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 607..616 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 613..1320 /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(758..776) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1026..1045 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 3262..3383 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3299..3318) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3353..3372 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3390..3845) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3527..3548) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3739..3758 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3872..3976 /gene="bla" /label=AmpR promoter promoter 3978..4335 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4001..4021) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4186..4321 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4248..4267 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4370..5164 /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(4424..4443) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5034..5053 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5352..5371) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5396..5443 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5772..6360 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6261..6280 /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 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc 661 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc 721 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt 781 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc 841 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc 901 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc 961 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc 1021 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac 1081 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc 1141 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc 1201 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg 1261 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag 1321 tccggactca gatctcgagc tcaagcttgc atgaagaagt cttactcagg aggcacgcgg 1381 acctccagtg gccggctccg gaggcttggt gactccagtg gcccagcctt aaaaagatct 1441 tttgaggtcg aggaggtcga gacacccaac tccaccccac cccggagggt ccagactccc 1501 ctactccgag ccactgtggc cagctccacc cagaaattcc aggacctggg cgtgaagaac 1561 tcagaaccct cggcccgcca tgtggactcc ctaagccaac gctcccccaa ggcgtccctg 1621 cggagggtgg agctctcggg ccccaaggcg gccgagccgg tgtcccggcg cactgagctg 1681 tccattgaca tctcgtccaa gcaggtggag aacgccgggg ccatcggccc gtcccggttc 1741 gggctcaaga gggccgaggt gttgggccac aagacgccag aaccggcccc tcggaggacg 1801 gagatcacca tcgtcaaacc ccaggagtca gcccaccgga ggatggagcc ccctgcctcc 1861 aaggtccccg aggtgcccac tgcccctgcc accgacgcag cccccaagag ggtggagatc 1921 cagatgccca agcctgctga ggcgcccacc gcccccagcc cagcccagac cttggagaat 1981 tcagagcctg cccctgtgtc tcagctgcag agcaggctgg agcccaagcc ccagccccct 2041 gtggctgagg ctacaccccg gagccaggag gccactgagg cggctcccag ctgcgttggc 2101 gacatggccg acacccccag agatgccggg ctcaagcagg cgcctgcatc acggaacgag 2161 aaggccccgg tggacttcgg ctacgtgggg attgactcca tcctggagca gatgcgccgg 2221 aaggccatga agcagggctt cgagttcaac atcatggtgg tcgggcagag cggcttgggt 2281 aaatccacct taatcaacac cctcttcaaa tccaaaatca gccggaagtc ggtgcagccc 2341 acctcagagg agcgcatccc caagaccatc gagatcaagt ccatcacgca cgatattgag 2401 gagaaaggcg tccggatgaa gctgacagtg attgacacac cagggttcgg ggaccacatc 2461 aacaacgaga actgctggca gcccatcatg aagttcatca atgaccagta cgagaaatac 2521 ctgcaggagg aggtcaacat caaccgcaag aagcgcatcc cggacacccg cgtccactgc 2581 tgcctctact tcatccccgc caccggccac tccctcaggc ccctggacat cgagtttatg 2641 aaacgcctga gcaaggtggt caacatcgtc cctgtcatcg ccaaggcgga cacactcacc 2701 ctggaggaga gggtccactt caaacagcgg atcaccgcag acctgctgtc caacggcatc 2761 gacgtgtacc cccagaagga atttgatgag gactcggagg accggctggt gaacgagaag 2821 ttccgggaga tgatcccatt tgctgtggtg ggcagtgacc acgagtacca ggtcaacggc 2881 aagaggatcc ttgggaggaa gaccaagtgg ggtaccatcg aagttgaaaa caccacacac 2941 tgtgagtttg cctacctgcg ggaccttctc atcaggacgc acatgcagaa catcaaggac 3001 atcaccagca gcatccactt cgaggcgtac cgtgtgaagc gcctcaacga gggcagcagc 3061 gccatggcca acggcatgga ggagaaggag ccagaagccc cagagatgta ggtcgacggt 3121 accgcgggcc cgggatccac cggatctaga taactgatca taatcagcca taccacattt 3181 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa 3241 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc 3301 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 3361 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt 3421 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat 3481 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa 3541 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg 3601 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa 3661 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc 3721 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag 3781 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg 3841 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 3901 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 3961 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg 4021 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 4081 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 4141 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact 4201 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 4261 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 4321 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca 4381 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg 4441 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg 4501 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc 4561 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt 4621 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc 4681 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca 4741 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc 4801 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg 4861 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct 4921 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc 4981 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc 5041 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta 5101 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt 5161 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga 5221 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac 5281 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg 5341 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca 5401 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg 5461 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc 5521 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc 5581 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat 5641 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 5701 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 5761 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 5821 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 5881 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta 5941 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 6001 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 6061 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 6121 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 6181 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 6241 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 6301 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 6361 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 6421 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat //