LOCUS       sequence-362798-        6905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mCherry-tagged DnaJB1-PKA catalytic subunit fusion N-terminal 
            myristoylation site and H33Q DnaJB1 mutation to block Hsp70 binding;
            for mammalian expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6905)
  AUTHORS   Zhang JZ, Lu TW, Stolerman LM, Tenner B, Yang JR, Zhang JF, Falcke 
            M, Rangamani P, Taylor SS, Mehta S, Zhang J
  TITLE     Phase Separation of a PKA Regulatory Subunit Controls cAMP 
            Compartmentation and Oncogenic Signaling.
  JOURNAL   Cell. 2020 Sep 17;182(6):1531-1544.e15. doi: 
            10.1016/j.cell.2020.07.043. Epub 2020 Aug 25.
  PUBMED    32846158
REFERENCE   2  (bases 1 to 6905)
  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: 
            "10.1016/j.cell.2020.07.043"; journalName: "Cell"; date: 
            "2020-09-17- 17"; volume: "182"; issue: "6"; pages: "1531-1544.e15"
COMMENT     Sequence Label: pcDNA3.1(+)-Myr-DnaJB1(H33Q)-PKAcat-mCherry
FEATURES             Location/Qualifiers
     source          1..6905
                     /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"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2171..2881
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2316..2334)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2584..2603
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2913..2930)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2919..3143
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3189..3617
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3276..3295)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3486..3507
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3626..3646)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3631..3960
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3811..3946
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3873..3892
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4008..4055
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4074..4448
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4578..4699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4615..4634)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4669..4688
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4748..4764)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4748..4764)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4761..4783)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4772..4788
                     /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(4796..4802,4803..4820,4821..4826))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4841..4862
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4979..4996)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5150..5738)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5230..5249)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5909..6700,6701..6769))
                     /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     6532..6551
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6770..6874)
                     /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 ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttatgg gcaacgccgc cgccgccaag aagggtaaag actactacca
      961 gacgttgggc ctggcccgcg gcgcgtcgga cgaggagatc aagcgggcct accgccgcca
     1021 ggcgctgcgc taccagccgg acaagaacaa ggagcccggc gccgaggaga agttcaagga
     1081 gatcgctgag gcctacgacg tgctcagcga cccgcgcaag cgcgagatct tcgaccgcta
     1141 cggggaggaa gtgaaagaat tcttagccaa agccaaagaa gattttctta aaaaatggga
     1201 aagtcccgct cagaacacag cccacttgga tcagtttgaa cgaatcaaga ccctcggcac
     1261 gggctccttc gggcgggtga tgctggtgaa acacaaggag accgggaacc actatgccat
     1321 gaagatcctc gacaaacaga aggtggtgaa actgaaacag atcgaacaca ccctgaatga
     1381 aaagcgcatc ctgcaagctg tcaactttcc gttcctcgtc aaactcgagt tctccttcaa
     1441 ggacaactca aacttataca tggtcatgga gtacgtgccc ggcggggaga tgttctcaca
     1501 cctacggcgg atcggaaggt tcagtgagcc ccatgcccgt ttctacgcgg cccagatcgt
     1561 cctgaccttt gagtatctgc actcgctgga tctcatctac agggacctga agccggagaa
     1621 tctgctcatt gaccagcagg gctacattca tgtgacagac ttcggtttcg ccaagcgcgt
     1681 gaagggccgc acttggacct tgtgcggcac ccctgagtac ctggcccctg agattatcct
     1741 gagcaaaggc tacaacaagg ccgtggactg gtgggccctg ggggttctta tctatgaaat
     1801 ggccgctggc tacccgccct tcttcgcaga ccagcccatc cagatctatg agaagatcgt
     1861 ctctgggaag gtgcgcttcc cttcccactt cagctctgac ttgaaggacc tgctgcggaa
     1921 cctcctgcag gtagatctca ccaagcgctt tgggaacctc aagaatgggg tcaacgatat
     1981 caagaaccac aagtggtttg ccacaactga ctggattgcc atctaccaga ggaaggtgga
     2041 agctcccttc ataccaaagt ttaaaggccc tggggatacg agtaactttg acgactatga
     2101 ggaagaagaa atccgggtct ccatcaatga gaagtgtggc aaggagtttt ctgagtttgg
     2161 atccgggagc atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat
     2221 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
     2281 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
     2341 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta
     2401 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa
     2461 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
     2521 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga
     2581 cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc
     2641 cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca
     2701 ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
     2761 ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga
     2821 acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta
     2881 atctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     2941 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3001 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3061 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3121 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3181 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3241 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3301 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3361 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3421 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3481 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3541 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3601 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3661 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3721 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3781 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3841 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3901 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3961 aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat
     4021 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca
     4081 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
     4141 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
     4201 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc
     4261 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
     4321 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
     4381 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
     4441 aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4501 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4561 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4621 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4681 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4741 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4801 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4861 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4921 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4981 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5041 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5101 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5161 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5221 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5281 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5341 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5401 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5461 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5521 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5581 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5641 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5701 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5761 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5821 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5881 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5941 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6001 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6061 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6121 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6181 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6241 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6301 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6361 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6421 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6481 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6541 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6601 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6661 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6721 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6781 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6841 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6901 acgtc
//
