LOCUS       sequence-416697-        6565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of HA-tagged human PPM1H S211A.
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 S211A
FEATURES             Location/Qualifiers
     source          1..6565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(95..112)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(266..854)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(346..365)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1025..1816,1817..1885))
                     /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     1648..1667
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1886..1990)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2065..2084)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2256..2635
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2636..2839
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2790..2810
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2884..2910
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4471..4497
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4538..4555)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4544..4768
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4901..4920)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5052..5099
                     /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             5107..6129
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    6259..6380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6296..6315)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6350..6369
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6429..6445)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6429..6445)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6442..6464)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6453..6469
                     /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(6477..6483,6484..6501,6502..6507))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6522..6543
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
       61 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
      121 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      181 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      241 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      301 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      361 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      421 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      481 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      541 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
      601 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
      661 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
      721 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
      781 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
      841 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
      901 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
      961 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1021 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1081 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1141 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1201 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1261 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1321 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1381 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1441 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1501 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1561 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     1621 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1681 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     1741 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     1801 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     1861 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     1921 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     1981 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     2041 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     2101 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     2161 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     2221 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
     2281 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     2341 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     2401 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     2461 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc
     2521 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     2581 acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     2641 gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag
     2701 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     2761 aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga
     2821 ggtctatata agcagagctc gtttagtgaa ccgtcagaat tggccaccgg atctgccgcc
     2881 atgtacccat acgatgtgcc agattacgcc ggatccatgc tcactcgagt gaaatctgcc
     2941 gtggccaatt tcatgggcgg catcatggct ggcagctcag gctccgagca cggcggcggc
     3001 agctgcggag gctcggacct gcccctgcgt ttcccctacg ggcggccaga gttcctgggg
     3061 ctgtctcagg acgaggtgga gtgcagcgcc gaccacatcg cccgccccat cctcatcctc
     3121 aaggagactc ggcggctgcc ctgggccact ggctacgcag aggttatcaa tgccgggaag
     3181 agcacacaca atgaagacca agccagctgt gaggtgctca ctgtgaagaa gaaggcaggg
     3241 gccgtgacct caaccccaaa caggaactca tccaagagac ggtcctccct tcccaatggg
     3301 gaagggctgc agctgaagga gaactcggaa tccgagggtg tttcctgcca ctattggtcg
     3361 ctgtttgacg ggcacgcggg gtccggggcc gcggtggtgg cgtcacgcct gctgcagcac
     3421 cacatcacgg agcagctgca ggacatcgtg gacatcctga agaactccgc cgtcctgccc
     3481 cctacctgcc tgggggagga gcctgagaac acgcccgcca acagccggac tctgacccgg
     3541 gcagccgccc tgcgcggagg ggtgggggcc ccgggctccc ccagcacgcc ccccacacgc
     3601 ttctttaccg agaagaagat tccccatgag tgcctggtca tcggagcgct tgaaagtgca
     3661 ttcaaggaaa tggacctaca gatagaacga gagaggagtt catataatat atctggtggc
     3721 tgcacggccc tcattgtgat ttgccttttg gggaagctgt atgttgcaaa tgctggggat
     3781 agcagggcca taatcatcag aaatggagaa attatcccca tgtcttcaga atttaccccc
     3841 gagacggagc gccagcgact tcagtacctg gcattcatgc agcctcactt gctgggaaat
     3901 gagttcacac atttggagtt tccaaggaga gtacagagaa aggagcttgg aaagaagatg
     3961 ctctacaggg actttaatat gacaggctgg gcatacaaaa ccattgagga tgaggacttg
     4021 aagttccccc ttatatatgg agaaggcaag aaggcccggg taatggcaac tattggagtg
     4081 accaggggac ttggggacca tgacctgaag gtgcatgact ccaacatcta cattaaacca
     4141 ttcctgtctt cagctccaga ggtaagaatc tacgatcttt caaaatatga tcatggatca
     4201 gatgatgtgc tgatcttggc cactgatgga ctctgggacg ttttatcaaa tgaagaagta
     4261 gcagaagcaa tcactcagtt tcttcctaac tgtgatccag atgatcctca caggtacaca
     4321 ctggcagctc aggacctggt gatgcgtgcc cggggtgtgc tgaaggacag aggatggcgg
     4381 atatctaatg accgactggg ctcaggagac gacatttctg tatatgtcat tcctttaata
     4441 catggaaaca agctgtcatg agcggccgct tacccatacg atgtgccaga ttacgcctaa
     4501 gtcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     4561 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     4621 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     4681 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     4741 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     4801 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4861 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4921 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4981 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     5041 tcacgtacct agaagttcct attccgaagt tcctattctc tagaaagtat aggaacttcc
     5101 ttggccaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc
     5161 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc
     5221 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa
     5281 gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac
     5341 attggggagt tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg
     5401 ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg
     5461 gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa
     5521 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg
     5581 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat
     5641 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc
     5701 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag
     5761 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg
     5821 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg
     5881 ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt
     5941 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc
     6001 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat
     6061 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca
     6121 aaggaatagc acgtactacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     6181 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     6241 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     6301 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6361 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     6421 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6481 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6541 ttaattgcgt tgcgctcact gcccg
//
