LOCUS       sequence-416698-        6565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of HA-tagged human PPM1H S124A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6565)
  AUTHORS   Wondwossen M Yeshaw , Ayan Adhikari, Claire Y Chiang, Herschel S 
            Dhekne, Paulina S Wawro, Suzanne R Pfeffer
  TITLE     Localization of PPM1H phosphatase tunes Parkinson's disease-linked 
            LRRK2 kinase-mediated Rab GTPase phosphorylation and ciliogenesis
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Oct 31;120(44)
REFERENCE   2  (bases 1 to 6565)
  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"; journalName: "Proc Natl 
            Acad Sci U S A. 2023 Oct 31;120(44)"
COMMENT     Sequence Label: pCMV5D HA-PPM1H S124A
FEATURES             Location/Qualifiers
     source          1..6565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(130..718)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(210..229)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(889..1680,1681..1749))
                     /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     1512..1531
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1750..1854)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1929..1948)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2120..2499
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2500..2703
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2654..2674
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2748..2774
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4335..4361
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4402..4419)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4408..4632
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4765..4784)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4916..4963
                     /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             4971..5993
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    6123..6244
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6160..6179)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6214..6233
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6293..6309)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6293..6309)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6306..6328)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6317..6333
                     /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(6341..6347,6348..6365,6366..6371))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6386..6407
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6524..6541)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
       61 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
      121 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      181 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
      241 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
      301 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
      361 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
      421 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
      481 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
      541 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
      601 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
      661 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
      721 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
      781 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
      841 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
      901 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
      961 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     1021 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     1081 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     1141 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     1201 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     1261 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     1321 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     1381 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     1441 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     1501 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     1561 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     1621 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     1681 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     1741 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     1801 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     1861 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     1921 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     1981 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     2041 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     2101 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     2161 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     2221 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     2281 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     2341 gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat tgacgtcaat
     2401 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga ctttcctact
     2461 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     2521 accaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     2581 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaataac
     2641 cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     2701 gctcgtttag tgaaccgtca gaattggcca ccggatctgc cgccatgtac ccatacgatg
     2761 tgccagatta cgccggatcc atgctcactc gagtgaaatc tgccgtggcc aatttcatgg
     2821 gcggcatcat ggctggcagc tcaggctccg agcacggcgg cggcagctgc ggaggctcgg
     2881 acctgcccct gcgtttcccc tacgggcggc cagagttcct ggggctgtct caggacgagg
     2941 tggagtgcag cgccgaccac atcgcccgcc ccatcctcat cctcaaggag actcggcggc
     3001 tgccctgggc cactggctac gcagaggtta tcaatgccgg gaagagcaca cacaatgaag
     3061 accaagccag ctgtgaggtg ctcactgtga agaagaaggc aggggccgtg acctcaaccc
     3121 caaacaggaa ctcatccaag agacggtccg cccttcccaa tggggaaggg ctgcagctga
     3181 aggagaactc ggaatccgag ggtgtttcct gccactattg gtcgctgttt gacgggcacg
     3241 cggggtccgg ggccgcggtg gtggcgtcac gcctgctgca gcaccacatc acggagcagc
     3301 tgcaggacat cgtggacatc ctgaagaact ccgccgtcct gccccctacc tgcctggggg
     3361 aggagcctga gaacacgccc gccaacagcc ggactctgac ccgggcagcc tccctgcgcg
     3421 gaggggtggg ggccccgggc tcccccagca cgccccccac acgcttcttt accgagaaga
     3481 agattcccca tgagtgcctg gtcatcggag cgcttgaaag tgcattcaag gaaatggacc
     3541 tacagataga acgagagagg agttcatata atatatctgg tggctgcacg gccctcattg
     3601 tgatttgcct tttggggaag ctgtatgttg caaatgctgg ggatagcagg gccataatca
     3661 tcagaaatgg agaaattatc cccatgtctt cagaatttac ccccgagacg gagcgccagc
     3721 gacttcagta cctggcattc atgcagcctc acttgctggg aaatgagttc acacatttgg
     3781 agtttccaag gagagtacag agaaaggagc ttggaaagaa gatgctctac agggacttta
     3841 atatgacagg ctgggcatac aaaaccattg aggatgagga cttgaagttc ccccttatat
     3901 atggagaagg caagaaggcc cgggtaatgg caactattgg agtgaccagg ggacttgggg
     3961 accatgacct gaaggtgcat gactccaaca tctacattaa accattcctg tcttcagctc
     4021 cagaggtaag aatctacgat ctttcaaaat atgatcatgg atcagatgat gtgctgatct
     4081 tggccactga tggactctgg gacgttttat caaatgaaga agtagcagaa gcaatcactc
     4141 agtttcttcc taactgtgat ccagatgatc ctcacaggta cacactggca gctcaggacc
     4201 tggtgatgcg tgcccggggt gtgctgaagg acagaggatg gcggatatct aatgaccgac
     4261 tgggctcagg agacgacatt tctgtatatg tcattccttt aatacatgga aacaagctgt
     4321 catgagcggc cgcttaccca tacgatgtgc cagattacgc ctaagtcgag tctagagggc
     4381 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     4441 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     4501 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     4561 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     4621 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     4681 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4741 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4801 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4861 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt acctagaagt
     4921 tcctattccg aagttcctat tctctagaaa gtataggaac ttccttggcc aaaaagcctg
     4981 aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc
     5041 tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg
     5101 gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc
     5161 ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg gagttcagcg
     5221 agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg
     5281 aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg
     5341 ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca
     5401 ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg
     5461 tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg
     5521 ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc
     5581 tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt
     5641 cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc
     5701 agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt
     5761 atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg
     5821 atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg
     5881 ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac
     5941 tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa tagcacgtac
     6001 tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6061 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6121 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6181 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6241 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     6301 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     6361 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6421 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6481 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6541 tgcgctcggt cgttcggctg cggcg
//
