LOCUS       sequence-189229-        7676 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 7676)
  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 7676)
  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 HSPH2
FEATURES             Location/Qualifiers
     source          1..7676
                     /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(3628..3645)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3634..3858
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3991..4010)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4142..4189
                     /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             4197..5219
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4197..5219
                     /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    5349..5470
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5386..5405)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5440..5459
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5519..5535)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5519..5535)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5532..5554)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5543..5559
                     /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(5567..5573,5574..5591,5592..5597))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5612..5633
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5750..5767)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5921..6509)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6001..6020)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6680..7471,7472..7540))
                     /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     7303..7322
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7541..7645)
                     /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 ctagtccagt gtggtggaat tctgcagata tcaccatgtc ggtggtgggc atagacctgg
     1081 gcttccagag ctgctacgtc gctgtggccc gcgccggcgg catcgagact atcgctaatg
     1141 agtatagcga ccgctgcacg ccggcttgca tttcttttgg tcctaagaat cgttcaattg
     1201 gagcagcagc taaaagccag gtaatttcta atgcaaagaa cacagtccaa ggatttaaaa
     1261 gattccatgg ccgagcattc tctgatccat ttgtggaggc agaaaaatct aaccttgcat
     1321 atgatattgt gcagttgcct acaggattaa caggtataaa ggtgacatat atggaggaag
     1381 agcgaaattt taccactgag caagtgactg ccatgctttt gtccaaactg aaggagacag
     1441 ccgaaagtgt tcttaagaag cctgtagttg actgtgttgt ttcggttcct tgtttctata
     1501 ctgatgcaga aagacgatca gtgatggatg caacacagat tgctggtctt aattgcttgc
     1561 gattaatgaa tgaaaccact gcagttgctc ttgcatatgg aatctataag caggatcttc
     1621 ctgccttaga agagaaacca agaaatgtag tttttgtaga catgggccac tctgcttatc
     1681 aagtttctgt atgtgcattt aatagaggaa aactgaaagt tctggccact gcatttgaca
     1741 cgacattggg aggtagaaaa tttgatgaag tgctagtaaa tcacttctgt gaagaatttg
     1801 ggaagaaata caagctagac attaagtcca aaatccgtgc attattacga ctctctcagg
     1861 agtgtgagaa actcaagaaa ttgatgagtg caaatgcttc agatctccct ttgagcattg
     1921 aatgttttat gaatgatgtt gatgtatctg gaactatgaa tagaggcaaa tttctggaga
     1981 tgtgcaatga tctcttagct agagtggagc caccacttcg tagtgttttg gaacaaacca
     2041 agttaaagaa agaagatatt tatgcagtgg agatagttgg tggtgctaca cgaatccctg
     2101 cggtaaaaga gaagatcagc aaatttttcg gtaaagaact tagtacaaca ttaaatgctg
     2161 atgaagctgt cactcgaggc tgtgcattgc agtgtaccat cttatcgcct gctttcaaag
     2221 tcagagaatt ttctatcact gatgtagtac catatccaat atctctgaga tggaattctc
     2281 cagctgaaga agggtcaagt gactgtgaag tcttttccaa aaatcatgct gctcctttct
     2341 ctaaagttct tacattttat agaaaggaac ctttcactct tgaggcctac tacagctctc
     2401 ctcaggattt gccctatcca gatcctgcta tagctcagtt ttcagttcag aaagtcactc
     2461 ctcagtctga tggctccagt tcaaaagtga aagtcaaagt tcgagtaaat gtccatggca
     2521 ttttcagtgt gtccagtgca tctttagtgg aggttcacaa gtctgaggaa aatgaggagc
     2581 caatggaaac agatcagaat gcaaaggagg aagagaagat gcaagtggac caggaggaac
     2641 cacatgttga agagcaacag cagcagacac cagcagaaaa taaggcagag tctgaagaaa
     2701 tggagacctc tcaagctgga tccaaggata aaaagatgga ccaaccaccc caagccaaga
     2761 aggcaaaagt gaagaccagt actgtggacc tgccaatcga gaaccagcta ttatggcaga
     2821 tagacagaga gatgctcaac ttgtacattg aaaatgaggg taagatgatc atgcaggata
     2881 aactggagaa ggagcggaat gatgctaaga acgcagtgga ggaatatgtg tatgaaatga
     2941 gagacaagct tagtggtgaa tatgagaagt ttgtgagtga agatgatcgt aacagtttta
     3001 ctttgaaact ggaagatact gaaaattggt tgtatgagga tggagaagac cagccaaagc
     3061 aagtttatgt tgataagttg gctgaattaa aaaatctagg ccaacctatt aagatacgtt
     3121 tccaggaatc tgaagaacga ccaaaattat ttgaagaact agggaaacag atccaacagt
     3181 atatgaaaat aatcagctct ttcaaaaaca aggaggacca gtatgatcat ttggatgctg
     3241 ccgacatggc aaaggtagaa aaaagcacaa atgaagcaat ggagtggatg aataacaagc
     3301 taaatctgca gaacaagcag agtttgacca tggatccagt tgtcaagtca aaagagattg
     3361 aagctaaaat taaggagctg acaagtactt gtagccctat aatttcaaag cccaaaccca
     3421 aagtggaacc tccaaaagag gaacaaaaaa aatgcagagc agaatggacc agtggatgga
     3481 caaggagaca acccaggccc ccaggctgct gagcagggta cagacacagc tgtgccttcg
     3541 gattcagaca agaagcttcc tgaaatggac attgattgat tccgcggccg ctcgagtcta
     3601 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     3661 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     3721 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     3781 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     3841 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     3901 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3961 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4021 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4081 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtacct
     4141 agaagttcct attccgaagt tcctattctc tagaaagtat aggaacttcc ttggccaaaa
     4201 agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct
     4261 ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag
     4321 ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg
     4381 tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggaat
     4441 tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc
     4501 tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg gatgcgatcg
     4561 ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc
     4621 aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc
     4681 aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc
     4741 tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca
     4801 atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg
     4861 gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg
     4921 agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc
     4981 gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt
     5041 tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga
     5101 ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag
     5161 aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaatagc
     5221 acgtactacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5281 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5341 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5401 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5461 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5521 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5581 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5641 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5701 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5761 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5821 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5881 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5941 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6001 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6061 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6121 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6181 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6241 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6301 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6361 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6421 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6481 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6541 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6601 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6661 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6721 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6781 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6841 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6901 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6961 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7021 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7081 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7141 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7201 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7261 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7321 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7381 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7441 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7501 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7561 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7621 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
