LOCUS       sequence-104505-        5881 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HSPB8 (small HSP) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5881)
  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 5881)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1093/hmg/ddq398"; journalName: "Hum Mol Genet"; date: 
            "2010-12-1- 1"; volume: "19"; issue: "23"; pages: "4677-93"
COMMENT     Sequence Label: Frt-hspB8
FEATURES             Location/Qualifiers
     source          1..5881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(826..843)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    832..1056
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1189..1208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    1340..1387
                     /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             1395..2417
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             1395..2417
                     /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    2547..2668
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2584..2603)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2638..2657
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2717..2733)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2717..2733)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2730..2752)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2741..2757
                     /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(join(2765..2771,2772..2789,2790..2795))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2810..2831
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2948..2965)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3119..3707)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3199..3218)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3878..4669,4670..4738))
                     /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     4501..4520
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4739..4843)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4918..4937)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5109..5488
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5489..5692
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5643..5663
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    5694..5712
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5715..5733
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     5743..5767
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 tcgagaacca tggctgacgg tcagatgccc ttctcctgcc actacccaag ccgcctgcgc
       61 cgagacccct tccgggactc tcccctctcc tctcgcctgc tggatgatgg ctttggcatg
      121 gaccccttcc cagacgactt gacagcctct tggcccgact gggctctgcc tcgtctctcc
      181 tccgcctggc caggcaccct aaggtcgggc atggtgcccc ggggccccac tgccaccgcc
      241 aggtttgggg tgcctgccga gggcaggacc cccccaccct tccctgggga gccctggaaa
      301 gtgtgtgtga atgtgcacag cttcaagcca gaggagttga tggtgaagac caaagatgga
      361 tacgtggagg tgtctggcaa acatgaagag aaacagcaag aaggtggcat tgtttctaag
      421 aacttcacaa agaaaatcca gcttcctgca gaggtggatc ctgtgacagt atttgcctca
      481 ctttccccag agggtctgct gatcatcgaa gctccccagg tccctcctta ctcaacattt
      541 ggagagagca gtttcaacaa cgagcttccc caggacagcc aggaagtcac ctgtacctga
      601 gatgccagta ctggcccatc cttgttttgt ccccaaccct agggcttctc tgattccagg
      661 atacattact ttagctgaac tcagatttag tgcaagtaaa atgttagagg gtgcgggggt
      721 gaggactgac cacagattcc ctggatagtg tagtggtaga tttctccaca ggatagcgac
      781 gcgccaaaat cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt
      841 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
      901 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
      961 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     1021 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     1081 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     1141 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     1201 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     1261 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     1321 gtgatggttc acgtacctag aagttcctat tccgaagttc ctattctcta gaaagtatag
     1381 gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg
     1441 aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt
     1501 tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt
     1561 tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag
     1621 tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc cgtgcacagg
     1681 gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg
     1741 aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg
     1801 gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc
     1861 cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg
     1921 ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg
     1981 cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact
     2041 ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc
     2101 cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg
     2161 caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga
     2221 gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg
     2281 tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct
     2341 ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc
     2401 cgagggcaaa ggaatagcac gtactacgag atttcgattc caccgccgcc ttctatgaaa
     2461 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     2521 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     2581 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2641 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     2701 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     2761 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     2821 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     2881 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2941 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3001 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3061 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3121 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3181 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3241 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3301 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3361 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3421 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3481 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3541 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3601 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3661 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3721 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3781 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3841 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3901 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3961 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4021 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4081 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4141 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4201 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4261 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4321 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4381 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4441 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4501 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4561 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4621 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4681 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4741 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4801 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4861 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     4921 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     4981 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     5041 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     5101 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     5161 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     5221 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     5281 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     5341 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     5401 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     5461 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     5521 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     5581 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     5641 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctccctat
     5701 cagtgataga gatctcccta tcagtgatag agatcgtcga cgagctcgtt tagtgaaccg
     5761 tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg
     5821 atccagcctc cggactctag cgtttaaact taagcttggt accgagctcg gatccaaaac
     5881 c
//
