LOCUS Exported 5729 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of HSP1 (small HSP) in mammalian cells. ACCESSION . VERSION . KEYWORDS Frt-hspB1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5729) AUTHORS Vos MJ, Zijlstra MP, Kanon B, van Waarde-Verhagen MA, Brunt ER, Oosterveld-Hut HM, Carra S, Sibon OC, Kampinga HH TITLE HSPB7 is the most potent polyQ aggregation suppressor within the HSPB family of molecular chaperones. JOURNAL Hum Mol Genet. 2010 Dec 1;19(23):4677-93. doi: 10.1093/hmg/ddq398. Epub 2010 Sep 15. PUBMED 20843828 REFERENCE 2 (bases 1 to 5729) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5729 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(674..691) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 680..904 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(1037..1056) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 1188..1235 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS 1243..2265 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " polyA_signal 2395..2516 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2432..2451) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2486..2505 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(2565..2581) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2565..2581) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2578..2600) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2589..2605 /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(2613..2643) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2658..2679 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2796..2813) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2967..3555) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3047..3066) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3726..4586) /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 4349..4368 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4587..4691) /gene="bla" /label=AmpR promoter primer_bind complement(4766..4785) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 4957..5336 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 5337..5540 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5491..5511 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 5542..5560 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 5563..5581 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 5591..5615 /label=LNCX /note="Human CMV promoter, forward primer" ORIGIN 1 tcgagagcat gaccgagcgc cgcgtcccct tctcgctcct gcggggcccc agctgggacc 61 ccttccgcga ctggtacccg catagccgcc tcttcgacca ggccttcggg ctgccccggc 121 tgccggagga gtggtcgcag tggttaggcg gcagcagctg gccaggctac gtgcgccccc 181 tgccccccgc cgccatcgag agccccgcag tggccgcgcc cgcctacagc cgcgcgctca 241 gccggcaact cagcagcggg gtctcggaga tccggcacac tgcggaccgc tggcgcgtgt 301 ccctggatgt caaccacttc gccccggacg agctgacggt caagaccaag gatggcgtgg 361 tggagatcac cggcaagcac gaggagcggc aggacgagca tggctacatc tcccggtgct 421 tcacgcggaa atacacgctg ccccccggtg tggaccccac ccaagtttcc tcctccctgt 481 cccctgaggg cacactgacc gtggaggccc ccatgcccaa gctagccacg cagtccaacg 541 agatcaccat cccagtcacc ttcgagtcgc gggcccagct tgggggccca gaagctgcaa 601 aatccgatga gactgccgcc aagtaaacgc gccaaaatcg agtctagagg gcccgtttaa 661 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc 721 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag 781 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag 841 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct 901 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt 961 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc 1021 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc 1081 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg 1141 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtacctagaa gttcctattc 1201 cgaagttcct attctctaga aagtatagga acttccttgg ccaaaaagcc tgaactcacc 1261 gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag 1321 ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc 1381 ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt 1441 gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag cgagagcctg 1501 acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa 1561 ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt 1621 agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg 1681 cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac 1741 gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac 1801 tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac 1861 aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac 1921 gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc 1981 tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc 2041 cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct 2101 tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca 2161 caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat 2221 agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagcacgt actacgagat 2281 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc 2341 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg 2401 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa 2461 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat 2521 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct 2581 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt 2641 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc 2701 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg 2761 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg 2821 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 2881 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 2941 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 3001 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 3061 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 3121 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat 3181 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 3241 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 3301 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 3361 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt 3421 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 3481 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 3541 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 3601 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 3661 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 3721 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 3781 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 3841 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 3901 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 3961 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 4021 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct 4081 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 4141 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 4201 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 4261 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 4321 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa 4381 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 4441 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 4501 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 4561 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 4621 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 4681 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc 4741 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt 4801 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta 4861 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg 4921 cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt 4981 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat 5041 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa 5101 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg 5161 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc 5221 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct 5281 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga 5341 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa 5401 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc 5461 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg 5521 aggtctatat aagcagagct ctccctatca gtgatagaga tctccctatc agtgatagag 5581 atcgtcgacg agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt 5641 ttgacctcca tagaagacac cgggaccgat ccagcctccg gactctagcg tttaaactta 5701 agcttggtac cgagctcgga tccaaaacc //