LOCUS       sequence-350535-        5304 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant protein synthesis of human HSPA8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5304)
  AUTHORS   Ayoub CA, Wagner CS, Kuret J
  TITLE     Identification of gene networks mediating regional resistance to 
            tauopathy in late-onset Alzheimer's disease.
  JOURNAL   PLoS Genet. 2023 Mar 27;19(3):e1010681. doi: 
            10.1371/journal.pgen.1010681. eCollection 2023 Mar.
  PUBMED    36972319
REFERENCE   2  (bases 1 to 5304)
  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"; journalName: "PLoS 
            Genet."; date: "2023-03-27"; volume: "19(3):e1010681. doi"; pages: "
            10.1371/journal.pgen.1010681. eCollection 2023 Mar"
COMMENT     Sequence Label: pT7C-HSPA8
FEATURES             Location/Qualifiers
     source          1..5304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             92..109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             119..136
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     rep_origin      complement(2184..2696)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2378..2399)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2590..2609
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(2723..2741)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2809..2913
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2914..2982,2983..3774)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3132..3151)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3945..4533
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4434..4453
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4687..4704
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4719..4858
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4784..4803)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4944..4966
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5202..5221
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacca tgggcagcag ccatcatcat catcatcaca gcagcggcct
      121 ggtgccgcgc ggcagccata tgactagtat gtccaaggga cctgcagttg gtattgatct
      181 tggcaccacc tactcttgtg tgggtgtttt ccagcacgga aaagtcgaga taattgccaa
      241 tgatcaggga aaccgaacca ctccaagcta tgtcgccttt acggacactg aacggttgat
      301 cggtgatgcc gcaaagaatc aagttgcaat gaaccccacc aacacagttt ttgatgccaa
      361 acgtctgatt ggacgcagat ttgatgacgc tgttgtccag tctgatatga aacattggcc
      421 ctttatggtg gtgaatgatg ctggcaggcc caaggtccaa gtagaataca agggagagac
      481 caaaagcttc tatccagagg aggtgtcttc tatggttctg acaaagatga aggaaattgc
      541 agaagcctac cttgggaaga ctgttaccaa tgctgtggtc acagtgccag cttactttaa
      601 tgactctcag cgtcaggcta ccaaagatgc tggaactatt gctggtctca atgtacttag
      661 aattattaat gagccaactg ctgctgctat tgcttacggc ttagacaaaa aggttggagc
      721 agaaagaaac gtgctcatct ttgacctggg aggtggcact tttgatgtgt caatcctcac
      781 tattgaggat ggaatctttg aggtcaagtc tacagctgga gacacccact tgggtggaga
      841 agattttgac aaccgaatgg tcaaccattt tattgctgag tttaagcgca agcataagaa
      901 ggacatcagt gagaacaaga gagccgtaag acgcctccgt actgcttgtg aacgtgctaa
      961 gcgtaccctc tcttccagca cccaggccag tattgagatc gattctctct atgaaggaat
     1021 cgacttctat acctccatta cccgtgcccg atttgaagaa ctgaatgctg acctgttccg
     1081 tggcaccctg gacccagtag agaaagccct tcgagatgcc aaactagaca agtcacagat
     1141 tcatgatatt gtcctggttg gtggttctac tcgtatcccc aagattcaga agcttctcca
     1201 agacttcttc aatggaaaag aactgaataa gagcatcaac cctgatgaag ctgttgctta
     1261 tggtgcagct gtccaggcag ccatcttgtc tggagacaag tctgagaatg ttcaagattt
     1321 gctgctcttg gatgtcactc ctctttccct tggtattgaa actgctggtg gagtcatgac
     1381 tgtcctcatc aagcgtaata ccaccattcc taccaagcag acacagacct tcactaccta
     1441 ttctgacaac cagcctggtg tgcttattca ggtttatgaa ggcgagcgtg ccatgacaaa
     1501 ggataacaac ctgcttggca agttcgaact cacaggcata cctcctgcac cccgaggtgt
     1561 tcctcagatt gaagtcactt ttgacattga tgccaatggt atactcaatg tctctgctgt
     1621 ggacaagagt acgggaaaag agaacaagat tactatcact aatgacaagg gccgtttgag
     1681 caaggaggac attgaacgta tggtccagga agctgagaag tacaaagctg aagatgagaa
     1741 gcagagggac aaggtgtcat ccaagaattc acttgagtcc tatgccttca acatgaaagc
     1801 aactgttgaa gatgagaaac ttcaaggcaa gattaacgat gaggacaaac agaagattct
     1861 ggacaagtgt aatgaaatta tcaactggct tgataagaat cagactgctg agaaggaaga
     1921 atttgaacat caacagaaag agctggagaa agtctgcaac cccatcatca ccaagctgta
     1981 ccagagtgca ggaggcatgc caggaggaat gcctggggga tttcctggtg gtggagctcc
     2041 tccctctggt ggtgcttcct cagggcccac cattgaagag gttgattaaa agggcgaatt
     2101 ctgcagatat ccatcacact ggcggccgct cgagtgatcc ggctgctaac aaagcccgaa
     2161 aggaagctga gttggctgct gccaccccgg ttgataatca gaaaagcccc aaaaacagga
     2221 agattgtata agcaaatatt taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2281 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2341 aatcaaaaga atagaccgag atagggttga gtttgttcca gtttggaaca agagtccact
     2401 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2461 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2521 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     2581 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     2641 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtgggg
     2701 atcaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     2761 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2821 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2881 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     2941 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     3001 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     3061 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     3121 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     3181 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     3241 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     3301 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     3361 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     3421 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc
     3481 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     3541 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3601 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     3661 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3721 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     3781 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3841 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3901 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3961 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4021 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4081 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4141 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4201 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4261 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4321 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4381 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4441 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4501 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4561 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4621 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4681 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     4741 acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg
     4801 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     4861 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4921 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4981 accgaaacgc gcgaggcagt cgatcgacgc tctcccttat gcgactcctg cattaggaag
     5041 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
     5101 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     5161 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     5221 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     5281 atcgagatct cgatcccgcg aaat
//
