LOCUS       sequence-162062-        7742 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 7742)
  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 7742)
  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 HSPH1
FEATURES             Location/Qualifiers
     source          1..7742
                     /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(3694..3711)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3700..3924
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4057..4076)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4208..4255
                     /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             4263..5285
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4263..5285
                     /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    5415..5536
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5452..5471)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5506..5525
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5585..5601)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5585..5601)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5598..5620)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5609..5625
                     /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(5633..5639,5640..5657,5658..5663))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5678..5699
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5816..5833)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5987..6575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6067..6086)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6746..7537,7538..7606))
                     /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     7369..7388
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7607..7711)
                     /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 tgtcggtggt ggggttggac gtgggctcgc agagctgcta catcgcggta gcccgggccg
     1141 ggggcatcga gaccatcgcc aatgagttca gcgaccggtg caccccgtca gtcatatcat
     1201 ttggatcaaa aaatagaaca atcggagttg cagccaaaaa tcagcaaatc actcatgcaa
     1261 acaatacggt gtctaacttc aaaagatttc atggccgagc attcaatgac cccttcattc
     1321 aaaaggagaa ggaaaacttg agttacgatt tggttccatt gaaaaatggt ggagttggaa
     1381 taaaggtaat gtacatgggt gaagaacatc tatttagtgt ggagcagata acagccatgt
     1441 tgttgactaa gctgaaggaa actgctgaaa acagcctcaa gaaaccagta acagattgtg
     1501 ttatttcagt cccctccttc tttacagatg ctgagaggcg atctgtgtta gatgctgcac
     1561 agattgttgg cctaaactgt ttaagactta tgaatgacat gacagctgtt gctttgaatt
     1621 acggaattta taagcaggat ctcccaagcc tggatgagaa acctcggata gtggtttttg
     1681 ttgatatggg acattcagct tttcaagtgt ctgcttgtgc ttttaacaag ggaaaattga
     1741 aggtactggg aacagctttt gatcctttct taggaggaaa aaacttcgat gaaaagttag
     1801 tggaacattt ttgtgcagaa tttaaaacta agtacaagtt ggatgcaaaa tccaaaatac
     1861 gagcactcct acgtctgtat caggaatgtg aaaaactgaa aaagctaatg agctctaaca
     1921 gcacagacct tccactgaat atcgaatgct ttatgaatga taaagatgtt tccggaaaga
     1981 tgaacaggtc acaatttgaa gaactctgtg ctgaacttct gcaaaagata gaagtacccc
     2041 tttattcact gttggaacaa actcatctca aagtagaaga tgtgagtgca gttgagattg
     2101 ttggaggcgc tacacgaatt ccagctgtga aggaaagaat tgccaaattc tttggaaaag
     2161 atattagcac aacactcaat gcagatgaag cagtagccag aggatgtgca ttacagtgtg
     2221 caatactttc cccggcattt aaagttagag aattttccgt cacagatgca gttccttttc
     2281 caatatctct gatctggaac catgattcag aagatactga aggtgttcat gaagtcttta
     2341 gtcgaaacca tgctgctcct ttctccaaag ttctcacctt tctgagaagg gggccttttg
     2401 agctagaagc tttctattct gatccccaag gagttccata tccagaagca aaaataggcc
     2461 gctttgtagt tcagaatgtt tctgcacaga aagatggaga aaaatctaga gtaaaagtca
     2521 aagtgcgagt caacacccat ggcattttca ccatctctac ggcatctatg gtggagaaag
     2581 tcccaactga ggagaatgaa atgtcttctg aagctgacat ggagtgtctg aatcagagac
     2641 caccagaaaa cccagacact gataaaaatg tccagcaaga caacagtgaa gctggaacac
     2701 agccccaggt acaaactgat gctcaacaaa cctcacagtc tcccccttca cctgaactta
     2761 cctcagaaga aaacaaaatc ccagatgctg acaaagcaaa tgaaaaaaaa gttgaccagc
     2821 ctccagaagc taaaaagccc aaaataaagg tggtgaatgt tgagctgcct attgaagcca
     2881 acttggtctg gcagttaggg aaagaccttc ttaacatgta tattgagaca gagggtaaga
     2941 tgataatgca agataaattg gaaaaagaaa ggaatgatgc taaaaatgca gttgaggaat
     3001 atgtgtatga gttcagagac aagctgtgtg gaccatatga aaaatttata tgtgagcagg
     3061 atcatcaaaa ttttttgaga ctcctcacag aaactgaaga ctggctgtat gaagaaggag
     3121 aggaccaagc taaacaagca tatgttgaca agttggaaga attaatgaaa attggcactc
     3181 cagttaaagt tcggtttcag gaagctgaag aacggccaaa aatgtttgaa gaactaggac
     3241 agaggctgca gcattatgcc aagatagcag ctgacttcag aaataaggat gagaaataca
     3301 accatattga tgagtctgaa atgaaaaaag tggagaagtc tgttaatgaa gtgatggaat
     3361 ggatgaataa tgtcatgaat gctcaggcta aaaagagtct tgatcaggat ccagttgtac
     3421 gtgctcagga aattaaaaca aaaatcaagg aattgaacaa cacatgtgaa cccgttgtaa
     3481 cacaaccgaa accaaaaatt gaatcaccca aactggaaag aactccaaat ggcccaaata
     3541 ttgataaaaa ggaagaagat ttagaagaca aaaacaattt tggtgctgaa cctccacatc
     3601 agaatggtga atgttaccct aatgagaaaa attctgttaa tatggacttg gactagctcg
     3661 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     3721 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3781 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3841 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3901 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     3961 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4021 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4081 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4141 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4201 gtacctagaa gttcctattc cgaagttcct attctctaga aagtatagga acttccttgg
     4261 ccaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca
     4321 gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg
     4381 taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc
     4441 gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg
     4501 gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc
     4561 aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg
     4621 cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa
     4681 tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc
     4741 actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc
     4801 tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct
     4861 ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga
     4921 tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt
     4981 gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc
     5041 ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg
     5101 gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag
     5161 ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct
     5221 gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg
     5281 aatagcacgt actacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5341 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5401 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5461 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5521 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5581 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5641 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5701 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5761 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5821 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5881 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5941 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6001 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6061 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6121 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6181 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6241 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6301 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6361 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6421 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6481 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6541 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6601 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6661 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6721 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6781 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6841 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6901 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6961 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7021 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7081 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7141 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7201 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7261 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7321 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7381 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7441 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7501 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7561 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7621 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7681 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7741 tc
//
