LOCUS       sequence-9171-ex        7675 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 7675)
  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 7675)
  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 HSPH2
FEATURES             Location/Qualifiers
     source          1..7675
                     /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(3627..3644)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3633..3857
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3990..4009)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4141..4188
                     /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             4196..5218
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4196..5218
                     /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    5348..5469
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5385..5404)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5439..5458
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5518..5534)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5518..5534)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5531..5553)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5542..5558
                     /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(5566..5572,5573..5590,5591..5596))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5611..5632
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5749..5766)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5920..6508)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6000..6019)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6679..7470,7471..7539))
                     /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     7302..7321
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7540..7644)
                     /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 ggtggtgggc atagacctgg
     1081 gcttccagag ctgctacgtc gctgtggccc gcgccggcgg catcgagact atcgctaatg
     1141 agtatagcga ccgctgcacg ccggcttgca tttcttttgg tcctaagaat cgttcaattg
     1201 gagcagcagc taaaagccag gtaatttcta atgcaaagaa cacagtccaa ggatttaaaa
     1261 gattccatgg ccgagcattc tctgatccat ttgtggaggc agaaaaatct aaccttgcat
     1321 atgatattgt gcagttgcct acaggattaa caggtataaa ggtgacatat atggaggaag
     1381 agcgaaattt taccactgag caagtgactg ccatgctttt gtccaaactg aaggagacag
     1441 ccgaaagtgt tcttaagaag cctgtagttg actgtgttgt ttcggttcct tgtttctata
     1501 ctgatgcaga aagacgatca gtgatggatg caacacagat tgctggtctt aattgcttgc
     1561 gattaatgaa tgaaaccact gcagttgctc ttgcatatgg aatctataag caggatcttc
     1621 ctgccttaga agagaaacca agaaatgtag tttttgtaga catgggccac tctgcttatc
     1681 aagtttctgt atgtgcattt aatagaggaa aactgaaagt tctggccact gcatttgaca
     1741 cgacattggg aggtagaaaa tttgatgaag tgttagtaaa tcacttctgt gaagaatttg
     1801 ggaagaaata caagctagac attaagtcca aaatccgtgc attattacga ctctctcagg
     1861 agtgtgagaa actcaagaaa ttgatgagtg caaatgcttc agatctccct ttgagcattg
     1921 aatgttttat gaatgatgtt gatgtatctg gaactatgaa tagaggcaaa tttctggaga
     1981 tgtgcaatga tctcttagct agagtggagc caccacttcg tagtgttttg gaacaaacca
     2041 agttaaagaa agaagatatt tatgcagtgg agatagttgg tggtgctaca cgaatccctg
     2101 cggtaaaaga gaagatcagc aaatttttcg gtaaagaact tagtacaaca ttaaatgctg
     2161 atgaagctgt cactcgaggc tgtgcattgc agtgtgccat cttatcgcct gctttcaaag
     2221 tcagagaatt ttctatcact gatgtagtac catatccaat atctctgaga tggaattctc
     2281 cagctgaaga agggtcaagt gactgtgaag tcttttccaa aaatcatgct gctcctttct
     2341 ctaaagttct tacattttat agaaaggaac ctttcactct tgaggcctac tacagctctc
     2401 ctcaggattt gccctatcca gatcctgcta tagctcagtt ttcagttcag aaagtcactc
     2461 ctcagtctga tggctccagt tcaaaagtga aagtcaaagt tcgagtaaat gtccatggca
     2521 ttttcagtgt gtccagtgca tctttagtgg aggttcacaa gtctgaggaa aatgaggagc
     2581 caatggaaac agatcagaat gcaaaggagg aagagaagat gcaagtggac caggaggaac
     2641 cacatgttga agagcaacag cagcagacac cagcagaaaa taaggcagag tctgaagaaa
     2701 tggagacctc tcaagctgga tccaaggata aaaagatgga ccaaccaccc caagccaaga
     2761 aggcaaaagt gaagaccagt actgtggacc tgccaatcga gaatcagcta ttatggcaga
     2821 tagacagaga gatgctcaac ttgtacattg aaaatgaggg taagatgatc atgcaggata
     2881 aactggagaa ggagcggaat gatgctaaga acgcagtgga ggaatatgtg tatgaaatga
     2941 gagacaagct tagtggtgaa tatgagaagt ttgtgagtga agatgatcgt aacagtttta
     3001 ctttgaaact ggaagatact gaaaattggt tgtatgagga tggagaagac cagccaaagc
     3061 aagtttatgt tgataagttg gctgaattaa aaaatctagg tcaacctatt aagatacgtt
     3121 tccaggaatc tgaagaacga ccaaaattat ttgaagaact agggaaacag atccaacagt
     3181 atatgaaaat aatcagctct ttcaaaaaca aggaggacca gtatgatcat ttggatgctg
     3241 ctgacatgac aaaggtagaa aaaagcacaa atgaagcaat ggagtggatg aataacaagc
     3301 taaatctgca gaacaagcag agtttgacca tggatccagt tgtcaagtca aaagagattg
     3361 aagctaaaat taaggagctg acaagtactt gtagccctat aatttcaaag cccaaaccca
     3421 aagtggaacc tccaaaagag gaacaaaaaa atgcagagca gaatggacca gtggatggac
     3481 aaggagacaa cccaggcccc caggctgctg agcagggtac agacacagct gtgccttcgg
     3541 attcagacaa gaagcttcct gaaatggaca ttgattgatt ccgcggccgc tcgagtctag
     3601 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     3661 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3721 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3781 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     3841 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     3901 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3961 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4021 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4081 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtaccta
     4141 gaagttccta ttccgaagtt cctattctct agaaagtata ggaacttcct tggccaaaaa
     4201 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     4261 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     4321 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     4381 ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt
     4441 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     4501 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc
     4561 tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     4621 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     4681 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     4741 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     4801 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     4861 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     4921 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg
     4981 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     5041 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     5101 tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga
     5161 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagca
     5221 cgtactacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5281 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5341 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5401 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5461 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5521 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5581 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5641 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5701 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5761 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5821 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5881 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5941 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6001 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6061 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6121 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6181 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6241 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6301 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6361 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6421 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6481 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6541 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6601 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6661 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6721 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6781 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6841 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6901 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6961 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7021 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7081 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7141 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7201 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7261 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7321 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7381 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7441 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7501 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7561 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7621 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
