LOCUS       sequence-9099-ex        7048 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 7048)
  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 7048)
  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 DNAK
FEATURES             Location/Qualifiers
     source          1..7048
                     /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(3000..3017)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3006..3230
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3363..3382)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3514..3561
                     /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             3569..4591
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3569..4591
                     /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    4721..4842
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4758..4777)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4812..4831
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4891..4907)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4891..4907)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4904..4926)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4915..4931
                     /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(4939..4945,4946..4963,4964..4969))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4984..5005
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5122..5139)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5293..5881)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5373..5392)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6052..6843,6844..6912))
                     /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     6675..6694
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6913..7017)
                     /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 caccatgggt aaaataattg
     1021 gtatcgacct gggtactacc aactcttgtg tagcgattat ggatggcacc actcctcgcg
     1081 tgctggagaa cgccgaaggc gatcgcacca cgccttctat cattgcctat acccaggatg
     1141 gtgaaactct agttggtcag ccggctaaac gtcaggcagt gacgaacccg caaaacactc
     1201 tgtttgcgat taaacgcctg attggtcgcc gcttccagga cgaagaagta cagcgtgatg
     1261 tttccatcat gccgttcaaa attattgctg ctgataacgg cgacgcatgg gtcgaagtta
     1321 aaggccagaa aatggcaccg ccgcagattt ctgctgaagt gctgaaaaaa atgaagaaaa
     1381 ccgctgaaga ttacctgggt gaaccggtaa ctgaagctgt tatcaccgta ccggcatact
     1441 ttaacgatgc tcagcgtcag gcaaccaaag acgcaggccg tatcgctggt ctggaagtaa
     1501 aacgtatcat caacgaaccg accgcagctg cgctggctta cggtctggac aaaggcactg
     1561 gcaaccgtac tatcgcggtt tatgacctgg gtggtggtac tttcgatatt tctattatcg
     1621 aaatcgacga agttgacggc gaaaaaacct tcgaagttct ggcaaccaac ggtgataccc
     1681 acctgggggg tgaagacttc gacagccgtc tgatcaacta tctggttgaa gaattcaaga
     1741 aagatcaggg cattgacctg cgcaacgatc cgctggcaat gcagcgcctg aaagaagcgg
     1801 cagaaaaagc gaaaatcgaa ctgtcttccg ctcagcagac cgacgttaac ctgccataca
     1861 tcactgcaga cgcgaccggt ccgaaacaca tgaacatcaa agtgactcgt gcgaaactgg
     1921 aaagcctggt tgaagatctg gtaaaccgtt ccattgagcc gctgaaagtt gcactgcagg
     1981 acgctggcct gtccgtatct gatatcgacg acgttatcct cgttggtggt cagactcgta
     2041 tgccaatggt tcagaagaaa gttgctgagt tctttggtaa agagccgcgt aaagacgtta
     2101 acccggacga agctgtagca atcggtgctg ctgttcaggg tggtgttctg actggtgacg
     2161 taaaagacgt actgctgctg gacgttaccc cgctgtctct gggtatcgaa accatgggcg
     2221 gtgtgatgac gacgctgatc gcgaaaaaca ccactatccc gaccaagcac agccaggtgt
     2281 tctctaccgc tgaagacaac cagtctgcgg taaccatcca tgtgctgcag ggtgaacgta
     2341 aacgtgcggc tgataacaaa tctctgggtc agttcaacct agatggtatc aacccggcac
     2401 cgcgcggcat gccgcagatc gaagttacct tcgatatcga tgctgacggt atcctgcacg
     2461 tttccgcgaa agataaaaac agcggtaaag agcagaagat caccatcaag gcttcttctg
     2521 gtctgaacga agatgaaatc cagaaaatgg tacgcgacgc agaagctaac gccgaagctg
     2581 accgtaagtt tgaagagctg gtacagactc gcaaccaggg cgaccatctg ctgcacagca
     2641 cccgtaagca ggttgaagaa gcaggcgaca aactgccggc tgacgacaaa actgctatcg
     2701 agtctgcgct gactgcactg gaaactgctc tgaaaggtga agacaaagcc gctatcgaag
     2761 cgaaaatgca ggaactggca caggtttccc agaaactgat ggaaatcgcc cagcagcaac
     2821 atgcccagca gcagactgcc ggtgctgatg cttctgcaaa caacgcgaaa gatgacgatg
     2881 ttgtcgacgc tgaatttgaa gaagtcaaag acaaaaaata atcgccctat aaacgggtaa
     2941 ttatactgac acggggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc
     3001 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3061 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3121 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3181 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     3241 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     3301 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3361 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3421 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3481 aaaacttgat tagggtgatg gttcacgtac ctagaagttc ctattccgaa gttcctattc
     3541 tctagaaagt ataggaactt ccttggccaa aaagcctgaa ctcaccgcga cgtctgtcga
     3601 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
     3661 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
     3721 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct
     3781 cccgattccg gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc
     3841 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
     3901 gcagccggtc gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg
     3961 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
     4021 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc
     4081 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg
     4141 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac
     4201 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat
     4261 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag
     4321 gcatccggag cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga
     4381 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg
     4441 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag
     4501 aagcgcggcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg
     4561 ccccagcact cgtccgaggg caaaggaata gcacgtacta cgagatttcg attccaccgc
     4621 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4681 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4741 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4801 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     4861 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4921 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4981 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5041 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5101 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5161 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5221 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5281 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5341 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5401 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5461 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5521 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5581 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5641 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5701 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5761 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5821 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5881 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5941 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6001 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6061 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6121 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6181 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6241 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6301 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6361 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6421 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6481 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6541 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6601 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6661 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6721 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6781 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6841 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6901 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6961 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7021 atttccccga aaagtgccac ctgacgtc
//
