LOCUS       sequence-364042-        6716 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mCherry-tagged PKA catalytic subunit lacking N-terminal 
            myristoylation site; for mammalian expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6716)
  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 6716)
  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(+)-PKAcat(G1A)-mCherry
FEATURES             Location/Qualifiers
     source          1..6716
                     /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             1982..2692
                     /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(2127..2145)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2395..2414
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2724..2741)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2730..2954
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3000..3428
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3087..3106)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3297..3318
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3437..3457)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3442..3771
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3622..3757
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3684..3703
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3819..3866
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3885..4259
                     /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    4389..4510
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4426..4445)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4480..4499
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4559..4575)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4559..4575)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4572..4594)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4583..4599
                     /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(4607..4613,4614..4631,4632..4637))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4652..4673
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4790..4807)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4961..5549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5041..5060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5720..6511,6512..6580))
                     /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     6343..6362
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6581..6685)
                     /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 atgtccagta 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 ccaacgccgc cgccgccaag aagggcagcg agcaggagag
      961 cgtgaaagaa ttcttagcca aagccaaaga agattttctt aaaaaatggg aaagtcccgc
     1021 tcagaacaca gcccacttgg atcagtttga acgaatcaag accctcggca cgggctcctt
     1081 cgggcgggtg atgctggtga aacacaagga gaccgggaac cactatgcca tgaagatcct
     1141 cgacaaacag aaggtggtga aactgaaaca gatcgaacac accctgaatg aaaagcgcat
     1201 cctgcaagct gtcaactttc cgttcctcgt caaactcgag ttctccttca aggacaactc
     1261 aaacttatac atggtcatgg agtacgtgcc cggcggggag atgttctcac acctacggcg
     1321 gatcggaagg ttcagtgagc cccatgcccg tttctacgcg gccaagatcg tcctgacctt
     1381 tgagtatctg cactcgctgg atctcatcta cagggacctg aagccggaga atctgctcat
     1441 tgaccagcag ggctacattc aggtgacaga cttcggtttc gccaagcgcg tgaagggccg
     1501 cacttggacc ttgtgcggca cccctgagta cctggcccct gagattatcc tgagcaaagg
     1561 ctacaacaag gccgtggact ggtgggccct gggggttctt atctatgaaa tggccgctgg
     1621 ctacccgccc ttcttcgcag accagcccat ccagatctat gagaagatcg tctctgggaa
     1681 ggtgcgcttc ccttcccact tcagctctga cttgaaggac ctgctgcgga acctcctgca
     1741 ggtagatctc accaagcgct ttgggaacct caagaatggg gtcaacgata tcaagaacca
     1801 caagtggttt gccacaactg actggattgc catctaccag aggaaggtgg aagctccctt
     1861 cataccaaag tttaaaggcc ctggggatac gagtaacttt gacgactatg aggaagaaga
     1921 aatccgggtc tccatcaatg agaagtgtgg caaggagttt tctgagtttg gatccgggag
     1981 catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     2041 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     2101 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     2161 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     2221 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     2281 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     2341 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     2401 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     2461 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     2521 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     2581 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     2641 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt aatctagagg
     2701 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     2761 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2821 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     2881 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     2941 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3001 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3061 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3121 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3181 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3241 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3301 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3361 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3421 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3481 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3541 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3601 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3661 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3721 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     3781 ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat
     3841 agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc aagttgacca
     3901 gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc
     3961 ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg
     4021 tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg
     4081 tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact
     4141 tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt
     4201 tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac
     4261 acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4321 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4381 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4441 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4501 atgtatctta tcatgtctgt ataccgtcga cctctatcta gagcttggcg taatcatggt
     4561 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4621 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4681 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4741 gccaacgctc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4801 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4861 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4921 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4981 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5041 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5101 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5161 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5221 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt tagtccaacc
     5281 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5341 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5401 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5461 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5521 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5581 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5641 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5701 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5761 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5821 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5881 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5941 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6001 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6061 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6121 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6181 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6241 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6301 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6361 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6421 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6481 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6541 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6601 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6661 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
