LOCUS       sequence-9136-ex        6887 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 6887)
  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 6887)
  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 GFP DNAJB5
FEATURES             Location/Qualifiers
     source          1..6887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      6..15
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(12..14,15..17,18..728)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(12..14,15..17,18..728)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(57..78)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(318..337)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     665..686
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1861..1878)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1867..2091
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2224..2243)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2375..2422
                     /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             2430..3452
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             2430..3452
                     /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    3582..3703
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3619..3638)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3673..3692
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3752..3768)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3752..3768)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3765..3787)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3776..3792
                     /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(3800..3806,3807..3824,3825..3830))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3845..3866
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3983..4000)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4154..4742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4234..4253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4913..5704,5705..5773))
                     /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     5536..5555
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5774..5878)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5953..5972)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6144..6523
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6524..6727
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6678..6698
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    6729..6747
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6750..6768
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     6778..6802
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 agcttgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
       61 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
      121 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
      181 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
      241 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
      301 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
      361 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
      421 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
      481 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
      541 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
      601 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
      661 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
      721 tgtacaaggg atccactagt ccagtgtggt ggaattctgc agatatccag cacagtggcg
      781 gccgcaccat gggaaaagat tattacaaga ttcttgggat cccatcgggg gccaacgagg
      841 atgagatcaa gaaagcctac cggaagatgg ccttgaagta ccacccagac aagaataaag
      901 aacccaacgc tgaggagaag tttaaggaga ttgcagaggc ctatgatgtg ctaagtgacc
      961 ccaagaaacg gggcctgtat gaccagtatg gggaggaagg cctgaagacc ggcggtggca
     1021 catcaggtgg ctccagtggc tcctttcact acacctttca tggggacccc catgccacct
     1081 ttgcctcctt ctttggtggc tccaacccct tcgatatctt ctttgccagc agccgctcca
     1141 ctcggccctt cagtggcttt gacccagatg acatggatgt ggatgaagat gaggacccat
     1201 ttggcgcttt cggccgtttt ggcttcaatg ggctgagtag gggtccaagg cgagccccag
     1261 aaccactgta ccctcggcgc aaggtgcagg accccccagt ggtgcacgag ctgcgggtgt
     1321 ccctggagga gatctaccat ggctccacca agcgcatgaa gatcacaagg cgtcgcctca
     1381 accctgatgg gcgaactgtg cgcaccgagg acaagatcct gcacatagtc atcaagcgtg
     1441 gctggaagga aggcaccaag atcaccttcc ccaaagaagg cgacgccaca cctgacaaca
     1501 tccctgctga catcgtcttt gtgctcaaag acaagcccca tgcacacttc cgccgagatg
     1561 gcaccaacgt gctctacagt gccctgatca gcctcaagga ggcgctgtgt ggctgcactg
     1621 tgaacattcc cactatcgac ggccgagtga tccctttgcc ctgcaatgat gtcatcaagc
     1681 caggcaccgt gaagagactc cgtggggagg gccttccctt ccccaaagtg ccaactcagc
     1741 gaggagacct cattgttgag ttcaaagttc gcttcccaga cagattaaca ccacagacaa
     1801 gacagatcct taagcagcac ctaccctgtt cctaggggcc cgtttaaacc cgctgatcag
     1861 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     1921 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     1981 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2041 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     2101 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     2161 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     2221 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     2281 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     2341 aaaaacttga ttagggtgat ggttcacgta cctagaagtt cctattccga agttcctatt
     2401 ctctagaaag tataggaact tccttggcca aaaagcctga actcaccgcg acgtctgtcg
     2461 agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg
     2521 aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata
     2581 gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc
     2641 tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct
     2701 cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc
     2761 tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg
     2821 ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat
     2881 gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
     2941 cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
     3001 ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa
     3061 cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca
     3121 tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga
     3181 ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg
     3241 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc
     3301 gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca
     3361 gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac
     3421 gccccagcac tcgtccgagg gcaaaggaat agcacgtact acgagatttc gattccaccg
     3481 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     3541 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     3601 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3661 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     3721 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     3781 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     3841 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3901 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3961 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4021 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4081 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4141 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4201 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4261 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4321 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4381 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4441 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4501 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4561 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4621 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4681 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4741 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4801 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4861 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4921 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4981 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5041 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5101 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5161 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5221 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5281 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5341 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5401 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5461 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5521 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5581 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5641 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5701 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5761 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5821 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5881 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     5941 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     6001 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     6061 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     6121 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
     6181 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     6241 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     6301 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     6361 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     6421 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     6481 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     6541 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     6601 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     6661 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     6721 cagagctctc cctatcagtg atagagatct ccctatcagt gatagagatc gtcgacgagc
     6781 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
     6841 aagacaccgg gaccgatcca gcctccggac tctagcgttt aaactta
//
