LOCUS       sequence-162066-        7696 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 7696)
  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 7696)
  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 HSPH3
FEATURES             Location/Qualifiers
     source          1..7696
                     /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(3648..3665)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3654..3878
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4011..4030)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4162..4209
                     /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             4217..5239
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4217..5239
                     /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    5369..5490
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5406..5425)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5460..5479
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5539..5555)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5539..5555)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5552..5574)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5563..5579
                     /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(5587..5593,5594..5611,5612..5617))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5632..5653
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5770..5787)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5941..6529)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6021..6040)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6700..7491,7492..7560))
                     /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     7323..7342
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7561..7665)
                     /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 tgtctgtggt tggcattgac ctcggctttc tcaactgcta cattgctgtc gcgagaagtg
     1141 gcggcatcga gaccatcgcc aatgagtaca gcgacaggtg taccccggcc tgtatatcat
     1201 tgggatcaag aactcgagcc attggaaatg cagcaaagag ccagatagtc acgaacgtaa
     1261 gaaatacaat tcatggcttc aaaaagcttc atgggcgatc atttgatgat cccattgtgc
     1321 aaactgaaag gatcaggctt ccctatgaac tgcagaaaat gcctaatgga agtgcaggag
     1381 ttaaggtgcg gtacttagag gaagagagac cttttgcaat tgagcaagtt actggaatgc
     1441 tgttagccaa gcttaaagag acttcagaaa atgctttgaa gaaaccagtg gctgactgtg
     1501 tgatttcaat tcctagcttt tttactgatg ccgagagaag atctgtgatg gctgcagccc
     1561 aggttgcagg cttaaattgt ttaaggttga tgaatgaaac tactgcagtt gcactggcgt
     1621 atggaattta taaacaggat cttcccccat tagatgagaa accaagaaat gtagtattta
     1681 ttgatatggg acattctgcc tatcaggtct tggtttgtgc ttttaacaaa ggaaaactta
     1741 aagtcttggc tactaccttt gatccatatt tgggtggcag gaactttgat gaggctttag
     1801 tagactactt ctgtgatgag ttcaagacca aatataagat aaatgtgaaa gaaaactctc
     1861 gggccttgtt gcgtttatat caggaatgtg aaaaactaaa gaagctaatg agtgcaaatg
     1921 catcagatct tccattgaac attgagtgtt tcatgaatga ccttgatgtt tctagtaaaa
     1981 tgaacagggc tcaatttgaa caactgtgtg cttccctttt ggccagggtt gaaccacctt
     2041 taaaagcagt aatggaacaa gctaacttac aacgtgaaga cattagtagt atagaaattg
     2101 taggaggagc aacacgaatt cctgcagtga aagaacaaat cactaaattc tttcttaaag
     2161 acataagtac cacattaaat gctgatgaag ctgttgcaag aggatgtgcg ttacagtgtg
     2221 cgattctctc accagcattt aaagtgcgtg aattttccat aacagacctt gttccctatt
     2281 caatcacatt aaggtggaag acctcttttg aagatggaag tggggaatgt gaagttttct
     2341 gtaagaacca tcctgcccca ttctcaaaag tcattacttt ccacaagaag gaaccatttg
     2401 aactagaagc attttatact aatttacatg aagtgcctta tcctgatgca agaattggga
     2461 gcttcactat tcagaatgtt tttccacagt ctgatggtga tagttccaaa gtgaaggtta
     2521 aagttcgtgt taacatccat ggaatcttca gtgtggctag cgcatcagta attgagaagc
     2581 aaaatttgga aggcgatcac agtgatgctc caatggagac agaaacttca tttaaaaatg
     2641 aaaacaaaga taatatggat aaaatgcagg ttgatcaaga agaagggcat caaaaatgtc
     2701 atgctgaaca cactccagaa gaggaaattg atcatacagg agccaaaaca aagtcagctg
     2761 tctcagacaa acaagaccga ttaaatcaga cacttaaaaa aggaaaagtc aaaagtattg
     2821 atctaccgat ccagagtagc ctatgtagac aactaggcca agatcttctc aacagctaca
     2881 ttgaaaatga ggggaagatg atcatgcaag ataagttaga gaaagaaaga aatgatgcta
     2941 agaatgccgt tgaagaatat gtatatgatt ttagagacag gctgggcact gtctatgaaa
     3001 aattcatcac tccagaagac ttgagtaaac tgtctgcagt attggaagac acagaaaatt
     3061 ggctttatga agacggagag gaccaaccta aacaagttta tgtggataag cttcaagaac
     3121 taaagaaata cggccagcct attcaaatga agtacatgga gcatgaagag agaccaaaag
     3181 ccttaaatga cttgggaaaa aagatccaac ttgtcatgaa agtgatagaa gcttatagaa
     3241 acaaggatga aagatatgat catctggatc ctactgaaat ggaaaaggtt gaaaaatgta
     3301 tcagtgatgc catgagttgg ctgaatagta agatgaatgc acagaacaaa ctaagtctca
     3361 ctcaagatcc tgtggtaaaa gtttcagaaa tagtagcaaa gtcaaaggaa ctggataatt
     3421 tctgtaaccc catcatttac aagcccaaac caaaagcaga agttcctgaa gacaaaccaa
     3481 aagctaatag tgaacacaat ggcccaatgg atggacagag tggaactgaa actaaatcag
     3541 attcaacaaa agacagctca cagcatacta aatcctctgg agagatggaa gtggactaag
     3601 tctgcggccg ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     3661 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3721 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3781 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3841 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3901 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3961 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4021 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4081 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4141 gggtgatggt tcacgtacct agaagttcct attccgaagt tcctattctc tagaaagtat
     4201 aggaacttcc ttggccaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     4261 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     4321 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     4381 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
     4441 agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     4501 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     4561 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
     4621 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     4681 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     4741 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     4801 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     4861 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     4921 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     4981 tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     5041 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     5101 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt
     5161 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     5221 tccgagggca aaggaatagc acgtactacg agatttcgat tccaccgccg ccttctatga
     5281 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5341 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5401 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5461 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5521 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5581 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5641 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5701 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5761 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5821 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5881 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5941 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6001 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6061 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6121 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6181 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6241 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6301 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6361 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6421 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6481 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6541 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6601 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6661 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6721 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6781 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6841 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6901 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6961 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7021 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7081 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7141 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7201 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7261 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7321 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7381 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7441 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7501 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7561 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7621 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7681 agtgccacct gacgtc
//
