LOCUS       sequence-9153-ex        7090 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 7090)
  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 7090)
  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 DNAK
FEATURES             Location/Qualifiers
     source          1..7090
                     /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(3042..3059)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3048..3272
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3405..3424)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3556..3603
                     /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             3611..4633
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3611..4633
                     /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    4763..4884
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4800..4819)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4854..4873
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4933..4949)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4933..4949)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4946..4968)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4957..4973
                     /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(4981..4987,4988..5005,5006..5011))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5026..5047
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5164..5181)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5335..5923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5415..5434)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6094..6885,6886..6954))
                     /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     6717..6736
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6955..7059)
                     /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 tccaccatgg gtaaaataat tggtatcgac ctgggtacta
     1081 ccaactcttg tgtagcgatt atggatggca ccactcctcg cgtgctggag aacgccgaag
     1141 gcgatcgcac cacgccttct atcattgcct atacccagga tggtgaaact ctagttggtc
     1201 agccggctaa acgtcaggca gtgacgaacc cgcaaaacac tctgtttgcg attaaacgcc
     1261 tgattggtcg ccgcttccag gacgaagaag tacagcgtga tgtttccatc atgccgttca
     1321 aaattattgc tgctgataac ggcgacgcat gggtcgaagt taaaggccag aaaatggcac
     1381 cgccgcagat ttctgctgaa gtgctgaaaa aaatgaagaa aaccgctgaa gattacctgg
     1441 gtgaaccggt aactgaagct gttatcaccg taccggcata ctttaacgat gctcagcgtc
     1501 aggcaaccaa agacgcaggc cgtatcgctg gtctggaagt aaaacgtatc atcaacgaac
     1561 cgaccgcagc tgcgctggct tacggtctgg acaaaggcac tggcaaccgt actatcgcgg
     1621 tttatgacct gggtggtggt actttcgata tttctattat cgaaatcgac gaagttgacg
     1681 gcgaaaaaac cttcgaagtt ctggcaacca acggtgatac ccacctgggg ggtgaagact
     1741 tcgacagccg tctgatcaac tatctggttg aagaattcaa gaaagatcag ggcattgacc
     1801 tgcgcaacga tccgctggca atgcagcgcc tgaaagaagc ggcagaaaaa gcgaaaatcg
     1861 aactgtcttc cgctcagcag accgacgtta acctgccata catcactgca gacgcgaccg
     1921 gtccgaaaca catgaacatc aaagtgactc gtgcgaaact ggaaagcctg gttgaagatc
     1981 tggtaaaccg ttccattgag ccgctgaaag ttgcactgca ggacgctggc ctgtccgtat
     2041 ctgatatcga cgacgttatc ctcgttggtg gtcagactcg tatgccaatg gttcagaaga
     2101 aagttgctga gttctttggt aaagagccgc gtaaagacgt taacccggac gaagctgtag
     2161 caatcggtgc tgctgttcag ggtggtgttc tgactggtga cgtaaaagac gtactgctgc
     2221 tggacgttac cccgctgtct ctgggtatcg aaaccatggg cggtgtgatg acgacgctga
     2281 tcgcgaaaaa caccactatc ccgaccaagc acagccaggt gttctctacc gctgaagaca
     2341 accagtctgc ggtaaccatc catgtgctgc agggtgaacg taaacgtgcg gctgataaca
     2401 aatctctggg tcagttcaac ctagatggta tcaacccggc accgcgcggc atgccgcaga
     2461 tcgaagttac cttcgatatc gatgctgacg gtatcctgca cgtttccgcg aaagataaaa
     2521 acagcggtaa agagcagaag atcaccatca aggcttcttc tggtctgaac gaagatgaaa
     2581 tccagaaaat ggtacgcgac gcagaagcta acgccgaagc tgaccgtaag tttgaagagc
     2641 tggtacagac tcgcaaccag ggcgaccatc tgctgcacag cacccgtaag caggttgaag
     2701 aagcaggcga caaactgccg gctgacgaca aaactgctat cgagtctgcg ctgactgcac
     2761 tggaaactgc tctgaaaggt gaagacaaag ccgctatcga agcgaaaatg caggaactgg
     2821 cacaggtttc ccagaaactg atggaaatcg cccagcagca acatgcccag cagcagactg
     2881 ccggtgctga tgcttctgca aacaacgcga aagatgacga tgttgtcgac gctgaatttg
     2941 aagaagtcaa agacaaaaaa taatcgccct ataaacgggt aattatactg acacggggcg
     3001 gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     3061 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3121 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3181 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     3241 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     3301 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3361 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3421 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3481 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3541 tggttcacgt acctagaagt tcctattccg aagttcctat tctctagaaa gtataggaac
     3601 ttccttggcc aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     3661 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     3721 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     3781 caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct
     3841 tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     3901 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     3961 catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     4021 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     4081 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     4141 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     4201 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     4261 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     4321 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     4381 atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     4441 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     4501 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac
     4561 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     4621 ggcaaaggaa tagcacgtac tacgagattt cgattccacc gccgccttct atgaaaggtt
     4681 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     4741 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     4801 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4861 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     4921 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4981 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5041 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5101 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5161 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5221 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5281 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5341 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5401 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5461 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5521 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5581 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5641 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5701 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5761 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5821 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5881 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5941 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6001 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6061 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6121 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6181 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6241 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6301 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6361 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6421 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6481 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6541 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6601 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6661 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6721 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6781 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6841 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6901 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6961 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7021 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7081 acctgacgtc
//
