LOCUS Exported 5317 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Hsp27 expression plasmid. ACCESSION . VERSION . KEYWORDS pFLAG-CMV2-Hsp27-S78D/S82D SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5317) TITLE Moens lab Hsp27 plasmids REFERENCE 2 (bases 1 to 5317) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5317 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 45..67 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 59..76 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 60..76 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" enhancer 237..616 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 617..820 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 771..791 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 854..877 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" polyA_signal 1577..2199 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(1776..1795) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" primer_bind complement(2223..2243) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2228..2557 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2408..2543 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2470..2489 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(2595..2614) /label=T7 /note="T7 promoter, forward primer" promoter complement(2596..2614) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2628..2644) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2628..2644) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2641..2663) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2652..2668 /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(2676..2706) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2721..2742 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2859..2876) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3030..3618) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3110..3129) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3789..4649) /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 4412..4431 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4650..4754) /gene="bla" /label=AmpR promoter rep_origin 4781..5236 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4868..4887) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5078..5099 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 61 taaaacgacg gccagtgcca agctgatcta tacattgaat caatattggc aattagccat 121 attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta 181 tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttgaca 241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata 301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 481 gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 541 ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt 601 catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg gatagcggtt 661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggta 721 ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg 781 cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagaa 841 ttgatctacc atggactaca aagacgatga cgacaagctt gcggccgcga attcatcgat 901 agatctgata tcggtaccag tcgactctag aggatccagc atgaccgagc gccgcgtccc 961 cttctcgctc ctgcggggcc ccagctggga ccccttccgc gactggtacc cgcatagccg 1021 cctcttcgac caggccttcg ggctgccccg gctgccggag gagtggtcgc agtggttagg 1081 cggcagcagc tggccaggct acgtgcgccc cctgcccccc gccgccatcg agagccccgc 1141 agtggccgcg cccgcctaca gccgcgcgct cgatcggcaa ctcgatagcg gggtctcgga 1201 gatccggcac actgcggacc gctggcgcgt gtccctggat gtcaaccact tcgccccgga 1261 cgagctgacg gtcaagacca aggatggcgt ggtggagatc accggcaagc acgaggagcg 1321 gcaggacgag catggctaca tctcccggtg cttcacgcgg aaatacacgc tgccccccgg 1381 tgtggacccc acccaagttt cctcctccct gtcccctgag ggcacactga ccgtggaggc 1441 ccccatgccc aagctagcca cgcagtccaa cgagatcacc atcccagtca ccttcgagtc 1501 gcgggcccag cttgggggcc cagaagctgc aaaatccgat gagactgccg ccaagtaaag 1561 ccttagcccg gatcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct 1621 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt 1681 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg 1741 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc 1801 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc 1861 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt 1921 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca 1981 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc 2041 cttccctgtc cttctgattt taaaataact ataccagcag gaggacgtcc agacacagca 2101 taggctacct ggccatgccc aaccggtggg acatttgagt tgcttgcttg gcactgtcct 2161 ctcatgcgtt gggtccactc agtagatgcc tgttgaattg ggtacgcggc cagcttggct 2221 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 2281 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc 2341 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac 2401 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact 2461 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta 2521 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa 2581 ccagaaagtt aattccctat agtgagtcgt attaaattcg taatcatgtc atagctgttt 2641 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 2701 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 2761 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 2821 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 2881 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 2941 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 3001 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 3061 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 3121 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 3181 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 3241 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 3301 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 3361 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 3421 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 3481 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 3541 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 3601 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 3661 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 3721 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 3781 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 3841 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 3901 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 3961 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 4021 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 4081 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 4141 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 4201 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 4261 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 4321 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 4381 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 4441 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 4501 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 4561 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 4621 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 4681 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 4741 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca 4801 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 4861 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 4921 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 4981 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 5041 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 5101 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 5161 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata 5221 ttaacgctta caatttccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg 5281 tgcgggcctc ttcgctatta cgccagctgg cgaaagg //