LOCUS       sequence-162063-        7700 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 7700)
  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 7700)
  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..7700
                     /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(3652..3669)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3658..3882
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4015..4034)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4166..4213
                     /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             4221..5243
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4221..5243
                     /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    5373..5494
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5410..5429)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5464..5483
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5543..5559)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5543..5559)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5556..5578)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5567..5583
                     /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(5591..5597,5598..5615,5616..5621))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5636..5657
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5774..5791)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5945..6533)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6025..6044)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6704..7495,7496..7564))
                     /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     7327..7346
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7565..7669)
                     /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 gaagtgctag
     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 tcgagaacca 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 taggccaacc 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 aaaaaatgca
     3481 gagcagaatg gaccagtgga tggacaagga gacaacccag gcccccaggc tgctgagcag
     3541 ggtacagaca cagctgtgcc ttcggattca gacaagaagc ttcctgaaat ggacattgat
     3601 tgattccgcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg
     3661 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3721 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3781 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3841 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     3901 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     3961 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4021 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4081 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4141 attagggtga tggttcacgt acctagaagt tcctattccg aagttcctat tctctagaaa
     4201 gtataggaac ttccttggcc aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc
     4261 tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc
     4321 gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg
     4381 atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc
     4441 cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg
     4501 cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg
     4561 tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc
     4621 cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg
     4681 ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg
     4741 cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg
     4801 tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca
     4861 ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct
     4921 ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg
     4981 agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct
     5041 atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg
     5101 caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg
     5161 ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca
     5221 ctcgtccgag ggcaaaggaa tagcacgtac tacgagattt cgattccacc gccgccttct
     5281 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5341 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5401 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5461 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5521 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5581 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5641 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5701 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5761 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5821 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5881 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5941 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6001 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6061 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6121 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6181 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6241 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6301 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6361 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6421 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6481 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6541 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6601 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6661 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6721 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6781 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6841 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6901 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6961 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7021 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7081 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7141 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7201 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7261 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7321 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7381 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7441 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7501 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7561 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7621 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7681 gaaaagtgcc acctgacgtc
//
