LOCUS Exported 5317 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Hsp27 expression plasmid. ACCESSION . VERSION . KEYWORDS pFLAG-CMV2-Hsp27-S15A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5317) TITLE Moens lab plasmids REFERENCE 2 (bases 1 to 5317) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 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 126..148 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 140..157 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 141..157 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" enhancer 318..697 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 698..901 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 852..872 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 935..958 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" polyA_signal 1658..2280 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(1857..1876) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" primer_bind complement(2304..2324) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2309..2638 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2489..2624 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2551..2570 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(2676..2695) /label=T7 /note="T7 promoter, forward primer" promoter complement(2677..2695) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2709..2725) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2709..2725) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2722..2744) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2733..2749 /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(2757..2787) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2802..2823 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2940..2957) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3111..3699) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3191..3210) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3870..4730) /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 4493..4512 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4731..4835) /gene="bla" /label=AmpR promoter rep_origin 4862..5317 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4949..4968) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5159..5180 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa 181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta 241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc 301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg 361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc 421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga 601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg 661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac 721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt 781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc 841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc 901 tcgtttagtg aaccgtcaga attgatctac catggactac aaagacgatg acgacaagct 961 tgcggccgcg aattcatcga tagatctgat atcggtacca gtcgactcta gaggatccag 1021 catgaccgag cgccgcgtcc ccttctcgct cctgcggggc cccgcttggg accccttccg 1081 cgactggtac ccgcatagcc gcctcttcga ccaggccttc gggctgcccc ggctgccgga 1141 ggagtggtcg cagtggttag gcggcagcag ctggccaggc tacgtgcgcc ccctgccccc 1201 cgccgccatc gagagccccg cagtggccgc gcccgcctac agccgcgcgc tcagccggca 1261 actcagcagc ggggtctcgg agatccggca cactgcggac cgctggcgcg tgtccctgga 1321 tgtcaaccac ttcgccccgg acgagctgac ggtcaagacc aaggatggcg tggtggagat 1381 caccggcaag cacgaggagc ggcaggacga gcatggctac atctcccggt gcttcacgcg 1441 gaaatacacg ctgccccccg gtgtggaccc cacccaagtt tcctcctccc tgtcccctga 1501 gggcacactg accgtggagg cccccatgcc caagctagcc acgcagtcca acgagatcac 1561 catcccagtc accttcgagt cgcgggccca gcttgggggc ccagaagctg caaaatccga 1621 tgagactgcc gccaagtaaa gccttagccc ggatcccggg tggcatccct gtgacccctc 1681 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat 1741 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag 1801 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta 1861 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg 1921 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac 1981 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct 2041 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt 2101 acaggcgtga accactgctc ccttccctgt ccttctgatt ttaaaataac tataccagca 2161 ggaggacgtc cagacacagc ataggctacc tggccatgcc caaccggtgg gacatttgag 2221 ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact cagtagatgc ctgttgaatt 2281 gggtacgcgg ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg 2341 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg 2401 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 2461 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca 2521 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 2581 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa 2641 gctcctcgag gaactgaaaa accagaaagt taattcccta tagtgagtcg tattaaattc 2701 gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac 2761 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca 2821 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat 2881 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 2941 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 3001 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 3061 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 3121 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 3181 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 3241 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 3301 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 3361 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 3421 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 3481 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 3541 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 3601 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 3661 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 3721 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 3781 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 3841 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 3901 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 3961 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 4021 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 4081 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 4141 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 4201 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 4261 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 4321 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 4381 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 4441 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 4501 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 4561 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 4621 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 4681 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 4741 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 4801 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 4861 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 4921 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 4981 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt 5041 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg 5101 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt 5161 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta 5221 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt 5281 aacgcgaatt ttaacaaaat attaacgctt acaattt //