LOCUS       sequence-104506-        5592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HSPB9 (small HSP) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5592)
  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 5592)
  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-hspB9
FEATURES             Location/Qualifiers
     source          1..5592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(537..554)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    543..767
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(900..919)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    1051..1098
                     /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             1106..2128
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             1106..2128
                     /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    2258..2379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2295..2314)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2349..2368
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2428..2444)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2428..2444)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2441..2463)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2452..2468
                     /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(2476..2482,2483..2500,2501..2506))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2521..2542
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2659..2676)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2830..3418)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2910..2929)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3589..4380,4381..4449))
                     /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     4212..4231
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4450..4554)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4629..4648)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4820..5199
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5200..5403
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5354..5374
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    5405..5423
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5426..5444
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     5454..5478
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 tcgagaacca tgcagagagt cggtaacacc ttctccaacg agagccgggt ggcatcccgg
       61 tgtcccagcg tgggccttgc tgaacggaac cgggtggcca caatgccggt gcggctgctc
      121 agggacagtc cagcggctca ggaggacaat gaccatgcca gagacggttt ccaaatgaag
      181 ctggatgccc acggcttcgc cccggaggaa ctggtggtgc aggtggatgg ccaatggctg
      241 atggtgaccg gacagcagca actggacgtc agggacccgg aaagggtcag ttaccgcatg
      301 tcacagaagg tgcaccggaa aatgctcccg tccaacctga gtcctaccgc catgacctgc
      361 tgcctgaccc cctccgggca gctgtgggtc agaggccagt gtgtggcgct ggccctccct
      421 gaagcccaaa caggaccgtc cccgagactc gggagcctcg gctctaaggc ttccaacctg
      481 acccggtaaa cgcgccaaaa tcgagtctag agggcccgtt taaacccgct gatcagcctc
      541 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
      601 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
      661 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
      721 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
      781 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
      841 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
      901 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
      961 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     1021 acttgattag ggtgatggtt cacgtaccta gaagttccta ttccgaagtt cctattctct
     1081 agaaagtata ggaacttcct tggccaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     1141 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     1201 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     1261 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc
     1321 gattccggaa gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg
     1381 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     1441 gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt
     1501 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     1561 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
     1621 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     1681 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     1741 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     1801 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     1861 tccggagctt gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca
     1921 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     1981 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     2041 cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     2101 cagcactcgt ccgagggcaa aggaatagca cgtactacga gatttcgatt ccaccgccgc
     2161 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     2221 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     2281 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2341 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     2401 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     2461 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     2521 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2581 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2641 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2701 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2761 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2821 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2881 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2941 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3001 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3061 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3121 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3181 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3241 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3301 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3361 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3421 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3481 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3541 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3601 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3661 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3721 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3781 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3841 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3901 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3961 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4021 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4081 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4141 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4201 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4261 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4321 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4381 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4441 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4501 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4561 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     4621 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     4681 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     4741 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     4801 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     4861 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     4921 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     4981 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     5041 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     5101 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     5161 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     5221 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     5281 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     5341 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     5401 gctctcccta tcagtgatag agatctccct atcagtgata gagatcgtcg acgagctcgt
     5461 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
     5521 caccgggacc gatccagcct ccggactcta gcgtttaaac ttaagcttgg taccgagctc
     5581 ggatccaaaa cc
//
