LOCUS       sequence-9140-ex        6507 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 6507)
  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 6507)
  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 DNAJB8
FEATURES             Location/Qualifiers
     source          1..6507
                     /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(1481..1498)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1487..1711
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1844..1863)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    1995..2042
                     /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             2050..3072
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             2050..3072
                     /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    3202..3323
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3239..3258)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3293..3312
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3372..3388)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3372..3388)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3385..3407)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3396..3412
                     /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(3420..3426,3427..3444,3445..3450))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3465..3486
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3603..3620)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3774..4362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3854..3873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4533..5324,5325..5393))
                     /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     5156..5175
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5394..5498)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5573..5592)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5764..6143
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6144..6347
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6298..6318
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    6349..6367
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6370..6388
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     6398..6422
                     /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 atccaccatg gctaactact acgaagtgct gggcgtgcag gccagcgctt
      781 ccccggagga catcaagaaa gcctaccgca agctggccct tcgttggcac cccgacaaga
      841 accctgacaa taaggaggag gcggagaaga agttcaagct ggtgtctgag gcctatgagg
      901 ttctgtctga ctccaagaaa cgctccctgt atgaccgtgc tggctgtgac agctggcggg
      961 ctggtggcgg ggccagcacg ccctaccaca gccccttcga caccggctac accttccgta
     1021 accctgagga catcttccgg gagtttttcg gtggcctgga ccctttctcc tttgagttct
     1081 gggacagccc attcaatagt gaccgtggtg gccggggcca tggcctgagg ggggccttct
     1141 cggcaggctt tggagaattt ccggccttca tggaggcctt ctcatccttc aacatgctgg
     1201 gctgcagcgg gggcagccac accaccttct catccacctc cttcgggggc tccagttctg
     1261 gcagctcggg gttcaagtcg gtgatgtcgt ccaccgagat gatcaatggc cacaaggtca
     1321 ccaccaagcg catcgtggag aacgggcagg agcgcgtgga ggtggaggaa gacgggcagc
     1381 tcaagtcggt gactgtgaac ggcaaggagc agctcaaatg gatggacagc aagtaggcgg
     1441 ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     1501 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     1561 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     1621 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     1681 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     1741 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     1801 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     1861 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     1921 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     1981 ggttcacgta cctagaagtt cctattccga agttcctatt ctctagaaag tataggaact
     2041 tccttggcca aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag
     2101 ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc
     2161 ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac
     2221 aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt
     2281 gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc
     2341 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc
     2401 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg
     2461 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat
     2521 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc
     2581 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat
     2641 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc
     2701 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg
     2761 ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga
     2821 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg
     2881 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga
     2941 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc
     3001 gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg
     3061 gcaaaggaat agcacgtact acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3121 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     3181 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     3241 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3301 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     3361 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3421 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3481 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3541 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3601 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3661 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3721 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3781 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3841 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3901 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3961 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4021 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4081 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4141 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4201 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4261 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4321 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4381 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4441 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4501 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4561 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4621 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4681 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4741 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4801 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4861 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4921 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4981 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5041 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5101 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5161 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5221 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5281 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5341 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5401 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5461 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5521 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     5581 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     5641 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     5701 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     5761 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     5821 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     5881 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     5941 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     6001 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     6061 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     6121 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     6181 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     6241 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     6301 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc cctatcagtg
     6361 atagagatct ccctatcagt gatagagatc gtcgacgagc tcgtttagtg aaccgtcaga
     6421 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca
     6481 gcctccggac tctagcgttt aaactta
//
