LOCUS       sequence-104498-        5729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HSP1 (small HSP) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 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-hspB1
FEATURES             Location/Qualifiers
     source          1..5729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     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(join(2613..2619,2620..2637,2638..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(join(3726..4517,4518..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
//
