LOCUS       Exported                5730 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 5730)
  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 5730)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5730
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(675..692)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    681..905
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1038..1057)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    1189..1236
                     /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             1244..2266
                     /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    2396..2517
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2433..2452)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2487..2506
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2566..2582)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2566..2582)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2579..2601)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2590..2606
                     /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(2614..2644)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2659..2680
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2797..2814)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2968..3556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3048..3067)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3727..4587)
                     /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     4350..4369
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4588..4692)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4767..4786)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4958..5337
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5338..5541
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5492..5512
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    5543..5561
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5564..5582
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     5592..5616
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 tcgagaagca tgaccgagcg ccgcgtcccc ttctcgctcc tgcggggccc cagctgggac
       61 cccttccgcg actggtaccc gcatagccgc ctcttcgacc aggccttcgg gctgccccgg
      121 ctgccggagg agtggtcgca gtggttaggc ggcagcagct ggccaggcta cgtgcgcccc
      181 ctgccccccg ccgccatcga gagccccgca gtggccgcgc ccgcctacag ccgcgcgctc
      241 agccggcaac tcagcagcgg ggtctcggag atccggcaca ctgcggaccg ctggcgcgtg
      301 tccctggatg tcaaccactt cgccccggac gagctgacgg tcaagaccaa ggatggcgtg
      361 gtggagatca ccggcaagca cgaggagcgg caggacgagc atggctacat ctcccggtgc
      421 ttcacgcgga aatacacgct gccccccggt gtggacccca cccaagtttc ctcctccctg
      481 tcccctgagg gcacactgac cgtggaggcc cccatgccca agctagccac gcagtccaac
      541 gagatcacca tcccagtcac cttcgagtcg cgggcccagc ttgggggccc agaagctgca
      601 aaatccgatg agactgccgc caagtaaacg cgccaaaatc gagtctagag ggcccgttta
      661 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
      721 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
      781 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
      841 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
      901 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
      961 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     1021 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     1081 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     1141 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtacctaga agttcctatt
     1201 ccgaagttcc tattctctag aaagtatagg aacttccttg gccaaaaagc ctgaactcac
     1261 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     1321 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
     1381 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
     1441 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct
     1501 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
     1561 actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct
     1621 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     1681 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     1741 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     1801 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     1861 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     1921 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     1981 ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct
     2041 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     2101 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     2161 acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga
     2221 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagcacg tactacgaga
     2281 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     2341 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     2401 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     2461 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     2521 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     2581 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     2641 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2701 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2761 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2821 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2881 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2941 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3001 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3061 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3121 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3181 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3241 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3301 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3361 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3421 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3481 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3541 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3601 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3661 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3721 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3781 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3841 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3901 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3961 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4021 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4081 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4141 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4201 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4261 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4321 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4381 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4441 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4501 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4561 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4621 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4681 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     4741 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4801 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     4861 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
     4921 gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat
     4981 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     5041 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     5101 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     5161 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     5221 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     5281 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     5341 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     5401 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     5461 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     5521 gaggtctata taagcagagc tctccctatc agtgatagag atctccctat cagtgataga
     5581 gatcgtcgac gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt
     5641 tttgacctcc atagaagaca ccgggaccga tccagcctcc ggactctagc gtttaaactt
     5701 aagcttggta ccgagctcgg atccaaaacc
//
