LOCUS       sequence-9170-ex        7718 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 7718)
  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 7718)
  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 HSPH1
FEATURES             Location/Qualifiers
     source          1..7718
                     /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(3670..3687)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3676..3900
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4033..4052)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4184..4231
                     /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             4239..5261
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4239..5261
                     /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    5391..5512
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5428..5447)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5482..5501
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5561..5577)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5561..5577)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5574..5596)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5585..5601
                     /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(5609..5615,5616..5633,5634..5639))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5654..5675
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5792..5809)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5963..6551)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6043..6062)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6722..7513,7514..7582))
                     /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     7345..7364
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7583..7687)
                     /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 ctagtccagt gtggtggaat tctgcagata tcaccatgtc ggtggtgggg ttggacgtgg
     1081 gctcgcagag ctgctacatc gcggtagccc gggccggggg catcgagacc atcgccaatg
     1141 agttcagcga ccggtgcacc ccgtcagtca tatcatttgg atcaaaaaat agaacaatcg
     1201 gagttgcagc caaaaatcag caaatcactc atgcaaacaa tacggtgtct aacttcaaaa
     1261 gatttcatgg ccgagcattc aatgacccct tcattcaaaa ggagaaggaa aacttgagtt
     1321 acgatttggt tccattgaaa aatggtggag ttggaataaa ggtaatgtac atgggtgaag
     1381 aacatctatt tagtgtggag cagataacag ccatgttgtt gactaagctg aaggaaactg
     1441 ctgaaaacag cctcaagaaa ccagtaacag attgtgttat ttcagtcccc tccttcttta
     1501 cagatgctga gaggcgatct gtgttagatg ctgcacagat tgttggccta aactgtttaa
     1561 gacttatgaa tgacatgaca gctgttgctt tgaattacgg aatttataag caggatctcc
     1621 caagcctgga tgagaaacct cggatagtgg tttttgttga tatgggacat tcagcttttc
     1681 aagtgtctgc ttgtgctttt aacaagggaa aattgaaggt actgggaaca gcttttgatc
     1741 ctttcttagg aggaaaaaac ttcgatgaaa agttagtgga acatttttgt gcagaattta
     1801 aaactaagta caagttggat gcaaaatcca aaatacgagc actcctacgt ctgtatcagg
     1861 aatgtgaaaa actgaaaaag ctaatgagct ctaacagcac agaccttcca ctgaatatcg
     1921 aatgctttat gaatgataaa gatgtttccg gaaagatgaa caggtcacaa tttgaagaac
     1981 tctgtgctga acttctgcaa aagatagaag taccccttta ttcactgttg gaacaaactc
     2041 atctcaaagt agaagatgtg agtgcagttg agattgttgg aggcgctaca cgaattccag
     2101 ctgtgaagga aagaattgcc aaattctttg gaaaagatat tagcacaaca ctcaatgcag
     2161 atgaagcagt agccagagga tgtgcattac agtgtgcaat actttccccg gcatttaaag
     2221 ttagagaatt ttccgtcaca gatgcagttc cttttccaat atctctgatc tggaaccatg
     2281 attcagaaga tactgaaggt gttcatgaag tctttagtcg aaaccatgct gctcctttct
     2341 ccaaagttct cacctttctg agaagggggc cttttgagct agaagctttc tattctgatc
     2401 cccaaggagt tccatatcca gaagcaaaaa taggccgctt tgtagttcag aatgtttctg
     2461 cacagaaaga tggagaaaaa tctagagtaa aagtcaaagt gcgagtcaac acccatggca
     2521 ttttcaccat ctctacggca tctatggtgg agaaagtccc aactgaggag aatgaaatgt
     2581 cttctgaagc tgacatggag tgtctgaatc agagaccacc agaaaaccca gacactgata
     2641 aaaatgtcca gcaagacaac agtgaagctg gaacacagcc ccaggtacaa actgatgctc
     2701 aacaaacctc acagtctccc ccttcacctg aacttacctc agaagaaaac aaaatcccag
     2761 atgctgacaa agcaaatgaa aaaaaagttg accagcctcc agaagctaaa aagcccaaaa
     2821 taaaggtggt gaatgttgag ctgcctattg aagccaactt ggtctggcag ttagggaaag
     2881 accttcttaa catgtatatt gagacagagg gtaagatgat aatgcaagat aaattggaaa
     2941 aagaaaggaa tgatgctaaa aatgcagttg aggaatatgt gtatgagttc agagacaagc
     3001 tgtgtggacc atatgaaaaa tttatatgtg agcaggatca tcaaaatttt ttgagactcc
     3061 tcacagaaac tgaagactgg ctgtatgaag aaggagagga ccaagctaaa caagcatatg
     3121 ttgacaagtt ggaagaatta atgaaaattg gcactccagt taaagttcgg tttcaggaag
     3181 ctgaagaacg gccaaaaatg tttgaagaac taggacagag gctgcagcat tatgccaaga
     3241 tagcagctga cttcagaaat aaggatgaga aatacaacca tattgatgag tctgaaatga
     3301 aaaaagtgga gaagtctgtt aatgaagtga tggaatggat gaataatgtc atgaatgctc
     3361 aggctaaaaa gagtcttgat caggatccag ttgtacgtgc tcaggaaatt aaaacaaaaa
     3421 tcaaggaatt gaacaacaca tgtgaacccg ttgtaacaca accgaaacca aaaattgaat
     3481 cacccaaact ggaaagaact ccaaatggcc caaatattga taaaaaggaa gaagatttag
     3541 aagacaaaaa caattttggt gctgaacctc cacatcagaa tggtgaatgt taccctaatg
     3601 agaaaaattc tgttaatatg gacttggact agctcgagtc tagagggccc gtttaaaccc
     3661 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     3721 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     3781 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     3841 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     3901 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     3961 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4021 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4081 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4141 tcgaccccaa aaaacttgat tagggtgatg gttcacgtac ctagaagttc ctattccgaa
     4201 gttcctattc tctagaaagt ataggaactt ccttggccaa aaagcctgaa ctcaccgcga
     4261 cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct
     4321 cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc
     4381 gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat
     4441 cggccgcgct cccgattccg gaagtgcttg acattgggga attcagcgag agcctgacct
     4501 attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc
     4561 ccgctgttct gcagccggtc gcggaggcca tggatgcgat cgctgcggcc gatcttagcc
     4621 agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg
     4681 atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca
     4741 ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc
     4801 ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg
     4861 gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg
     4921 tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact
     4981 tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat atgctccgca
     5041 ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg
     5101 cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa
     5161 tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc gccgatagtg
     5221 gaaaccgacg ccccagcact cgtccgaggg caaaggaata gcacgtacta cgagatttcg
     5281 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5341 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5401 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5461 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5521 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5581 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5641 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5701 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5761 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5821 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5881 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5941 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6001 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6061 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6121 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6181 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6241 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6301 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6361 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6421 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6481 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6541 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6601 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6661 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6721 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6781 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6841 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6901 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6961 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7021 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7081 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7141 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7201 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7261 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7321 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7381 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7441 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7501 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7561 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7621 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7681 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
