LOCUS       sequence-9091-ex        7654 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 7654)
  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 7654)
  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 HSPH3
FEATURES             Location/Qualifiers
     source          1..7654
                     /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"
     primer_bind     complement(3606..3623)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3612..3836
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3969..3988)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4120..4167
                     /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             4175..5197
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4175..5197
                     /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    5327..5448
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5364..5383)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5418..5437
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5497..5513)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5497..5513)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5510..5532)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5521..5537
                     /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(5545..5551,5552..5569,5570..5575))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5590..5611
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5728..5745)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5899..6487)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5979..5998)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6658..7449,7450..7518))
                     /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     7281..7300
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7519..7623)
                     /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 cttggtaccg agctcggatc cactagtcca gtgtggtgga
     1021 attctgcaga tatcaccatg tctgtggttg gcattgacct cggctttctc aactgctaca
     1081 ttgctgtcgc gagaagtggc ggcatcgaga ccatcgccaa tgagtacagc gacaggtgta
     1141 ccccggcctg tatatcattg ggatcaagaa ctcgagccat tggaaatgca gcaaagagcc
     1201 agatagtcac gaacgtaaga aatacaattc atggcttcaa aaagcttcat gggcgatcat
     1261 ttgatgatcc cattgtgcaa actgaaagga tcaggcttcc ctatgaactg cagaaaatgc
     1321 ctaatggaag tgcaggagtt aaggtgcggt acttagagga agagagacct tttgcaattg
     1381 agcaagttac tggaatgctg ttagccaagc ttaaagagac ttcagaaaat gctttgaaga
     1441 aaccagtggc tgactgtgtg atttcaattc ctagcttttt tactgatgcc gagagaagat
     1501 ctgtgatggc tgcagcccag gttgcaggct taaattgttt aaggttgatg aatgaaacta
     1561 ctgcagttgc actggcgtat ggaatttata aacaggatct tcccccatta gatgagaaac
     1621 caagaaatgt agtatttatt gatatgggac attctgccta tcaggtcttg gtttgtgctt
     1681 ttaacaaagg aaaacttaaa gtcttggcta ctacctttga tccatatttg ggtggcagga
     1741 actttgatga ggctttagta gactacttct gtgatgagtt caagaccaaa tataagataa
     1801 atgtgaaaga aaactctcgg gccttgttgc gtttatatca ggaatgtgaa aaactaaaga
     1861 agctaatgag tgcaaatgca tcagatcttc cattgaacat tgagtgtttc atgaatgacc
     1921 ttgatgtttc tagtaaaatg aacagggctc aatttgaaca actgtgtgct tcccttttgg
     1981 ccagggttga accaccttta aaagcagtaa tggaacaagc taacttacaa cgtgaagaca
     2041 ttagtagtat agaaattgta ggaggagcaa cacgaattcc tgcagtgaaa gaacaaatca
     2101 ctaaattctt tcttaaagac ataagtacca cattaaatgc tgatgaagct gttgcaagag
     2161 gatgtgcgtt acagtgtgcg attctctcac cagcatttaa agtgcgtgaa ttttccataa
     2221 cagaccttgt tccctattca atcacattaa ggtggaagac ctcttttgaa gatggaagtg
     2281 gggaatgtga agttttctgt aagaaccatc ctgccccatt ctcaaaagtc attactttcc
     2341 acaagaagga accatttgaa ctagaagcat tttatactaa tttacatgaa gtgccttatc
     2401 ctgatgcaag aattgggagc ttcactattc agaatgtttt tccacagtct gatggtgata
     2461 gttccaaagt gaaggttaaa gttcgtgtta acatccatgg aatcttcagt gtggctagcg
     2521 catcagtaat tgagaagcaa aatttggaag gcgatcacag tgatgctcca atggagacag
     2581 aaacttcatt taaaaatgaa aacaaagata atatggataa aatgcaggtt gatcaagaag
     2641 aagggcatca aaaatgtcat gctgaacaca ctccagaaga ggaaattgat catacaggag
     2701 ccaaaacaaa gtcagctgtc tcagacaaac aagaccgatt aaatcagaca cttaaaaaag
     2761 gaaaagtcaa aagtattgat ctaccgatcc agagtagcct atgtagacaa ctaggccaag
     2821 atcttctcaa cagctacatt gaaaatgagg ggaagatgat catgcaagat aagttagaga
     2881 aagaaagaaa tgatgctaag aatgccgttg aagaatatgt atatgatttt agagacaggc
     2941 tgggcactgt ctatgaaaaa ttcatcactc cagaagactt gagtaaactg tctgcagtat
     3001 tggaagacac agaaaattgg ctttatgaag acggagagga ccaacctaaa caagtttatg
     3061 tggataagct tcaagaacta aagaaatacg gccagcctat tcaaatgaag tacatggagc
     3121 atgaagagag accaaaagcc ttaaatgact tgggaaaaaa gatccaactt gtcatgaaag
     3181 tgatagaagc ttatagaaac aaggatgaaa gatatgatca tctggatcct actgaaatgg
     3241 aaaaggttga aaaatgtatc agtgatgcca tgagttggct gaatagtaag atgaatgcac
     3301 agaacaaact aagtctcact caagatcctg tggtaaaagt ttcagaaata gtagcaaagt
     3361 caaaggaact ggataatttc tgtaacccca tcatttacaa gcccaaacca aaagcagaag
     3421 ttcctgaaga caaaccaaaa gctaatagtg aacacaatgg cccaatggat ggacagagtg
     3481 gaactgaaac taaatcagat tcaacaaaag acagctcaca gcatactaaa tcctctggag
     3541 agatggaagt ggactaagtc tgcggccgct cgagtctaga gggcccgttt aaacccgctg
     3601 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3661 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3721 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3781 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     3841 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3901 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3961 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4021 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4081 ccccaaaaaa cttgattagg gtgatggttc acgtacctag aagttcctat tccgaagttc
     4141 ctattctcta gaaagtatag gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc
     4201 tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga
     4261 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt
     4321 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc
     4381 cgcgctcccg attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg
     4441 catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
     4501 tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac
     4561 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
     4621 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
     4681 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
     4741 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
     4801 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
     4861 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
     4921 gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg
     4981 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
     5041 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
     5101 ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
     5161 ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac gtactacgag atttcgattc
     5221 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5281 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5341 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5401 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5461 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5521 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5581 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5641 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5701 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5761 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5821 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5881 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5941 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6001 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6061 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6121 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6181 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6241 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6301 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6361 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6421 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6481 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6541 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6601 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6661 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6721 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6781 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6841 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6901 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6961 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7021 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7081 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7141 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7201 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7261 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7321 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7381 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7441 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7501 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7561 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7621 gcgcacattt ccccgaaaag tgccacctga cgtc
//
