LOCUS       sequence-9144-ex        7699 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 7699)
  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 7699)
  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 V5 HSPH2
FEATURES             Location/Qualifiers
     source          1..7699
                     /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..1037
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3651..3668)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3657..3881
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4014..4033)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4165..4212
                     /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             4220..5242
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4220..5242
                     /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    5372..5493
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5409..5428)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5463..5482
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5542..5558)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5542..5558)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5555..5577)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5566..5582
                     /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(5590..5596,5597..5614,5615..5620))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5635..5656
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5773..5790)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5944..6532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6024..6043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6703..7494,7495..7563))
                     /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     7326..7345
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7564..7668)
                     /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 cttgaattca ccatgggtaa gcctatccct aaccctctcc
     1021 tcggtctcga ttctacggga tccactagtc cagtgtggtg gaattctgca gatatcacca
     1081 tgtcggtggt gggcatagac ctgggcttcc agagctgcta cgtcgctgtg gcccgcgccg
     1141 gcggcatcga gactatcgct aatgagtata gcgaccgctg cacgccggct tgcatttctt
     1201 ttggtcctaa gaatcgttca attggagcag cagctaaaag ccaggtaatt tctaatgcaa
     1261 agaacacagt ccaaggattt aaaagattcc atggccgagc attctctgat ccatttgtgg
     1321 aggcagaaaa atctaacctt gcatatgata ttgtgcagtt gcctacagga ttaacaggta
     1381 taaaggtgac atatatggag gaagagcgaa attttaccac tgagcaagtg actgccatgc
     1441 ttttgtccaa actgaaggag acagccgaaa gtgttcttaa gaagcctgta gttgactgtg
     1501 ttgtttcggt tccttgtttc tatactgatg cagaaagacg atcagtgatg gatgcaacac
     1561 agattgctgg tcttaattgc ttgcgattaa tgaatgaaac cactgcagtt gctcttgcat
     1621 atggaatcta taagcaggat cttcctgcct tagaagagaa accaagaaat gtagtttttg
     1681 tagacatggg ccactctgct tatcaagttt ctgtatgtgc atttaataga ggaaaactga
     1741 aagttctggc cactgcattt gacacgacat tgggaggtag aaaatttgat gaagtgttag
     1801 taaatcactt ctgtgaagaa tttgggaaga aatacaagct agacattaag tccaaaatcc
     1861 gtgcattatt acgactctct caggagtgtg agaaactcaa gaaattgatg agtgcaaatg
     1921 cttcagatct ccctttgagc attgaatgtt ttatgaatga tgttgatgta tctggaacta
     1981 tgaatagagg caaatttctg gagatgtgca atgatctctt agctagagtg gagccaccac
     2041 ttcgtagtgt tttggaacaa accaagttaa agaaagaaga tatttatgca gtggagatag
     2101 ttggtggtgc tacacgaatc cctgcggtaa aagagaagat cagcaaattt ttcggtaaag
     2161 aacttagtac aacattaaat gctgatgaag ctgtcactcg aggctgtgca ttgcagtgtg
     2221 ccatcttatc gcctgctttc aaagtcagag aattttctat cactgatgta gtaccatatc
     2281 caatatctct gagatggaat tctccagctg aagaagggtc aagtgactgt gaagtctttt
     2341 ccaaaaatca tgctgctcct ttctctaaag ttcttacatt ttatagaaag gaacctttca
     2401 ctcttgaggc ctactacagc tctcctcagg atttgcccta tccagatcct gctatagctc
     2461 agttttcagt tcagaaagtc actcctcagt ctgatggctc cagttcaaaa gtgaaagtca
     2521 aagttcgagt aaatgtccat ggcattttca gtgtgtccag tgcatcttta gtggaggttc
     2581 acaagtctga ggaaaatgag gagccaatgg aaacagatca gaatgcaaag gaggaagaga
     2641 agatgcaagt ggaccaggag gaaccacatg ttgaagagca acagcagcag acaccagcag
     2701 aaaataaggc agagtctgaa gaaatggaga cctctcaagc tggatccaag gataaaaaga
     2761 tggaccaacc accccaagcc aagaaggcaa aagtgaagac cagtactgtg gacctgccaa
     2821 tcgagaatca gctattatgg cagatagaca gagagatgct caacttgtac attgaaaatg
     2881 agggtaagat gatcatgcag gataaactgg agaaggagcg gaatgatgct aagaacgcag
     2941 tggaggaata tgtgtatgaa atgagagaca agcttagtgg tgaatatgag aagtttgtga
     3001 gtgaagatga tcgtaacagt tttactttga aactggaaga tactgaaaat tggttgtatg
     3061 aggatggaga agaccagcca aagcaagttt atgttgataa gttggctgaa ttaaaaaatc
     3121 taggtcaacc tattaagata cgtttccagg aatctgaaga acgaccaaaa ttatttgaag
     3181 aactagggaa acagatccaa cagtatatga aaataatcag ctctttcaaa aacaaggagg
     3241 accagtatga tcatttggat gctgctgaca tgacaaaggt agaaaaaagc acaaatgaag
     3301 caatggagtg gatgaataac aagctaaatc tgcagaacaa gcagagtttg accatggatc
     3361 cagttgtcaa gtcaaaagag attgaagcta aaattaagga gctgacaagt acttgtagcc
     3421 ctataatttc aaagcccaaa cccaaagtgg aacctccaaa agaggaacaa aaaaatgcag
     3481 agcagaatgg accagtggat ggacaaggag acaacccagg cccccaggct gctgagcagg
     3541 gtacagacac agctgtgcct tcggattcag acaagaagct tcctgaaatg gacattgatt
     3601 gattccgcgg ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     3661 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3721 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3781 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3841 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3901 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3961 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4021 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4081 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4141 ttagggtgat ggttcacgta cctagaagtt cctattccga agttcctatt ctctagaaag
     4201 tataggaact tccttggcca aaaagcctga actcaccgcg acgtctgtcg agaagtttct
     4261 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg
     4321 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
     4381 tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc
     4441 ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc
     4501 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt
     4561 cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc
     4621 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
     4681 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc
     4741 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
     4801 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat
     4861 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
     4921 gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga
     4981 gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta
     5041 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc
     5101 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc
     5161 cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac
     5221 tcgtccgagg gcaaaggaat agcacgtact acgagatttc gattccaccg ccgccttcta
     5281 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5341 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5401 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5461 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5521 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5581 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5641 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5701 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5761 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5821 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5881 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5941 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6001 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6061 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6121 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6181 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6241 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6301 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6361 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6421 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6481 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6541 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6601 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6661 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6721 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6781 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6841 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6901 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6961 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7021 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7081 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7141 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7201 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7261 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7321 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7381 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7441 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7501 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7561 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7621 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7681 aaaagtgcca cctgacgtc
//
