LOCUS       sequence-9180-ex        7066 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 7066)
  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 7066)
  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 DNAK
FEATURES             Location/Qualifiers
     source          1..7066
                     /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(3018..3035)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3024..3248
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3381..3400)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3532..3579
                     /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             3587..4609
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3587..4609
                     /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    4739..4860
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4776..4795)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4830..4849
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4909..4925)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4909..4925)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4922..4944)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4933..4949
                     /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(4957..4963,4964..4981,4982..4987))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5002..5023
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5140..5157)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5311..5899)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5391..5410)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6070..6861,6862..6930))
                     /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     6693..6712
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6931..7035)
                     /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 ccatgggtaa aataattggt atcgacctgg gtactaccaa ctcttgtgta gcgattatgg
     1081 atggcaccac tcctcgcgtg ctggagaacg ccgaaggcga tcgcaccacg ccttctatca
     1141 ttgcctatac ccaggatggt gaaactctag ttggtcagcc ggctaaacgt caggcagtga
     1201 cgaacccgca aaacactctg tttgcgatta aacgcctgat tggtcgccgc ttccaggacg
     1261 aagaagtaca gcgtgatgtt tccatcatgc cgttcaaaat tattgctgct gataacggcg
     1321 acgcatgggt cgaagttaaa ggccagaaaa tggcaccgcc gcagatttct gctgaagtgc
     1381 tgaaaaaaat gaagaaaacc gctgaagatt acctgggtga accggtaact gaagctgtta
     1441 tcaccgtacc ggcatacttt aacgatgctc agcgtcaggc aaccaaagac gcaggccgta
     1501 tcgctggtct ggaagtaaaa cgtatcatca acgaaccgac cgcagctgcg ctggcttacg
     1561 gtctggacaa aggcactggc aaccgtacta tcgcggttta tgacctgggt ggtggtactt
     1621 tcgatatttc tattatcgaa atcgacgaag ttgacggcga aaaaaccttc gaagttctgg
     1681 caaccaacgg tgatacccac ctggggggtg aagacttcga cagccgtctg atcaactatc
     1741 tggttgaaga attcaagaaa gatcagggca ttgacctgcg caacgatccg ctggcaatgc
     1801 agcgcctgaa agaagcggca gaaaaagcga aaatcgaact gtcttccgct cagcagaccg
     1861 acgttaacct gccatacatc actgcagacg cgaccggtcc gaaacacatg aacatcaaag
     1921 tgactcgtgc gaaactggaa agcctggttg aagatctggt aaaccgttcc attgagccgc
     1981 tgaaagttgc actgcaggac gctggcctgt ccgtatctga tatcgacgac gttatcctcg
     2041 ttggtggtca gactcgtatg ccaatggttc agaagaaagt tgctgagttc tttggtaaag
     2101 agccgcgtaa agacgttaac ccggacgaag ctgtagcaat cggtgctgct gttcagggtg
     2161 gtgttctgac tggtgacgta aaagacgtac tgctgctgga cgttaccccg ctgtctctgg
     2221 gtatcgaaac catgggcggt gtgatgacga cgctgatcgc gaaaaacacc actatcccga
     2281 ccaagcacag ccaggtgttc tctaccgctg aagacaacca gtctgcggta accatccatg
     2341 tgctgcaggg tgaacgtaaa cgtgcggctg ataacaaatc tctgggtcag ttcaacctag
     2401 atggtatcaa cccggcaccg cgcggcatgc cgcagatcga agttaccttc gatatcgatg
     2461 ctgacggtat cctgcacgtt tccgcgaaag ataaaaacag cggtaaagag cagaagatca
     2521 ccatcaaggc ttcttctggt ctgaacgaag atgaaatcca gaaaatggta cgcgacgcag
     2581 aagctaacgc cgaagctgac cgtaagtttg aagagctggt acagactcgc aaccagggcg
     2641 accatctgct gcacagcacc cgtaagcagg ttgaagaagc aggcgacaaa ctgccggctg
     2701 acgacaaaac tgctatcgag tctgcgctga ctgcactgga aactgctctg aaaggtgaag
     2761 acaaagccgc tatcgaagcg aaaatgcagg aactggcaca ggtttcccag aaactgatgg
     2821 aaatcgccca gcagcaacat gcccagcagc agactgccgg tgctgatgct tctgcaaaca
     2881 acgcgaaaga tgacgatgtt gtcgacgctg aatttgaaga agtcaaagac aaaaaataat
     2941 cgccctataa acgggtaatt atactgacac ggggcggccg ctcgagtcta gagggcccgt
     3001 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3061 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3121 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3181 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3241 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3301 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3361 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3421 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3481 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtacct agaagttcct
     3541 attccgaagt tcctattctc tagaaagtat aggaacttcc ttggccaaaa agcctgaact
     3601 caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat
     3661 gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata
     3721 tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca
     3781 ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag
     3841 cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac
     3901 cgaactgccc gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga
     3961 tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac
     4021 atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat
     4081 ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga
     4141 ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac
     4201 ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca
     4261 atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac
     4321 gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat
     4381 gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc
     4441 agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg
     4501 tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc
     4561 cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaatagc acgtactacg
     4621 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4681 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4741 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4801 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4861 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4921 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4981 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5041 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5101 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5161 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5221 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5281 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5341 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5401 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5461 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5521 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5581 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5641 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5701 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5761 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5821 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5881 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5941 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6001 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6061 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6121 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6181 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6241 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6301 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6361 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6421 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6481 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6541 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6601 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6661 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6721 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6781 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6841 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6901 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6961 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7021 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
