LOCUS       sequence-9177-ex        7056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7056)
  AUTHORS   Hageman J, Kampinga HH
  TITLE     Computational analysis of the human HSPH/HSPA/DNAJ family and 
            cloning of a human HSPH/HSPA/DNAJ expression library.
  JOURNAL   Cell Stress Chaperones. 2009 Jan;14(1):1-21. doi: 
            10.1007/s12192-008-0060-2. Epub 2008 Aug 7.
  PUBMED    18686016
REFERENCE   2  (bases 1 to 7056)
  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: 
            "10.1007/s12192-008-0060-2"; journalName: "Cell Stress Chaperones"; 
            date: "2009-01"; volume: "14"; issue: "1"; pages: "1-21"
COMMENT     Sequence Label: pcDNA5/FRT/TO HIS HSPA8
FEATURES             Location/Qualifiers
     source          1..7056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             996..1013
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3008..3025)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3014..3238
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3371..3390)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3522..3569
                     /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             3577..4599
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3577..4599
                     /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    4729..4850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4766..4785)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4820..4839
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4899..4915)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4899..4915)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4912..4934)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4923..4939
                     /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(4947..4953,4954..4971,4972..4977))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4992..5013
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5130..5147)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5301..5889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5381..5400)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6060..6851,6852..6920))
                     /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     6683..6702
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6921..7025)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttctgcaga ccatgcatca tcaccatcac catggatcca
     1021 ccatgtccaa gggacctgca gttggtattg atcttggcac cacctactct tgtgtgggtg
     1081 ttttccagca cggaaaagtc gagataattg ccaatgatca gggaaaccga accactccaa
     1141 gctatgtcgc ctttacggac actgaacggt tgatcggtga tgccgcaaag aatcaagttg
     1201 caatgaaccc caccaacaca gtttttgatg ccaaacgtct gattggacgc agatttgatg
     1261 atgctgttgt ccagtctgat atgaaacatt ggccctttat ggtggtgaat gatgctggca
     1321 ggcccaaggt ccaagtagaa tacaagggag agaccaaaag cttctatcca gaggaggtgt
     1381 cttctatggt tctgacaaag atgaaggaaa ttgcagaagc ctaccttggg aagactgtta
     1441 ccaatgctgt ggtcacagtg ccagcttact ttaatgactc tcagcgtcag gctaccaaag
     1501 atgctggaac tattgctggt ctcaatgtac ttagaattat taatgagcca actgctgctg
     1561 ctattgctta cggcttagac aaaaaggttg gagcagaaag aaacgtgctc atctttgacc
     1621 tgggaggtgg cacttttgat gtgtcaatcc tcactattga ggatggaatc tttgaggtca
     1681 agtctacagc tggagacacc cacttgggtg gagaagattt tgacaaccga atggtcaacc
     1741 attttattgc tgagtttaag cgcaagcata agaaggacat cagtgagaac aagagagctg
     1801 taagacgcct ccgtactgct tgtgaacgtg ctaagcgtac cctctcttcc agcacccagg
     1861 ccagtattga gatcgattct ctctatgaag gaatcgactt ctatacctcc attacccgtg
     1921 cccgatttga agaactgaat gctgacctgt tccgtggcac cctggaccca gtagagaaag
     1981 cccttcgaga tgccaaacta gacaagtcac agattcatga tattgtcctg gttggtggtt
     2041 ctactcgtat ccccaagatt cagaagcttc tccaagactt cttcaatgga aaagaactga
     2101 ataagagcat caaccctgat gaagctgttg cttatggtgc agctgtccag gcagccatct
     2161 tgtctggaga caagtctgag aatgttcaag atttgctgct cttggatgtc actcctcttt
     2221 cccttggtat tgaaactgct ggtggagtca tgactgtcct catcaagcgt aataccacca
     2281 ttcctaccaa gcagacacag accttcacta cctattctga caaccagcct ggtgtgctta
     2341 ttcaggttta tgaaggcgag cgtgccatga caaaggataa caacctgctt ggcaagtttg
     2401 aactcacagg catacctcct gcaccccgag gtgttcctca gattgaagtc acttttgaca
     2461 ttgatgccaa tggtatactc aatgtctctg ctgtggacaa gagtacggga aaagagaaca
     2521 agattactat cactaatgac aagggccgtt tgagcaagga agacattgaa cgtatggtcc
     2581 aggaagctga gaagtacaaa gctgaagatg agaagcagag ggacaaggtg tcatccaaga
     2641 attcacttga gtcctatgcc ttcaacatga aagcaactgt tgaagatgag aaacttcaag
     2701 gcaagattaa cgatgaggac aaacagaaga ttctggacaa gtgtaatgaa attatcaact
     2761 ggcttgataa gaatcagact gctgagaagg aagaatttga acatcaacag aaagagctgg
     2821 agaaagtttg caaccccatc atcaccaagc tgtaccagag tgcaggaggc atgccaggag
     2881 gaatgcctgg gggatttcct ggtggtggag ctcctccctc tggtggtgct tcctcagggc
     2941 ccaccattga agaggttgat taagcggccg ctcgagtcta gagggcccgt ttaaacccgc
     3001 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3061 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3121 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3181 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3241 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3301 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3361 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3421 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3481 gaccccaaaa aacttgatta gggtgatggt tcacgtacct agaagttcct attccgaagt
     3541 tcctattctc tagaaagtat aggaacttcc ttggccaaaa agcctgaact caccgcgacg
     3601 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg
     3661 gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg
     3721 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg
     3781 gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat
     3841 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc
     3901 gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag
     3961 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat
     4021 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
     4081 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc
     4141 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc
     4201 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc
     4261 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc
     4321 gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt
     4381 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg
     4441 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc
     4501 gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga
     4561 aaccgacgcc ccagcactcg tccgagggca aaggaatagc acgtactacg agatttcgat
     4621 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4681 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4741 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4801 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4861 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4921 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4981 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5041 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5101 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5161 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5221 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5281 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5341 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5401 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5461 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5521 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5581 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5641 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5701 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5761 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5821 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5881 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5941 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6001 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6061 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6121 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6181 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6241 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6301 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6361 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6421 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6481 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6541 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6601 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6661 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6721 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6781 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6841 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6901 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6961 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7021 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
