LOCUS       sequence-412901-        6466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of HA-Delta-115-133, 204-217 PPM1H.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6466)
  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 6466)
  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-Delta-115-133, 204-217 PPM1H
FEATURES             Location/Qualifiers
     source          1..6466
                     /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"
     CDS             863..889
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2351..2377
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2418..2435)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2424..2648
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2781..2800)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2932..2979
                     /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             2987..4009
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4139..4260
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4176..4195)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4230..4249
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4309..4325)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4309..4325)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4322..4344)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4333..4349
                     /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(4357..4363,4364..4381,4382..4387))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4402..4423
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4540..4557)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4711..5299)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4791..4810)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5470..6261,6262..6330))
                     /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     6093..6112
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6331..6435)
                     /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 aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta cgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagaatt
      841 ggccaccgga tctgccgcca tgtacccata cgatgtgcca gattacgccg gatccatgct
      901 cactcgagtg aaatctgccg tggccaattt catgggcggc atcatggctg gcagctcagg
      961 ctccgagcac ggcggcggca gctgcggagg ctcggacctg cccctgcgtt tcccctacgg
     1021 gcggccagag ttcctggggc tgtctcagga cgaggtggag tgcagcgccg accacatcgc
     1081 ccgccccatc ctcatcctca aggagactcg gcggctgccc tgggccactg gctacgcaga
     1141 ggttatcaat gccgggaaga gcacacacaa tgaagaccaa gccagctgtg aggtgctcac
     1201 tgtgaagaag aaggcagggg ccgtgacctc aaccccaaag gagaactcgg aatccgaggg
     1261 tgtttcctgc cactattggt cgctgtttga cgggcacgcg gggtccgggg ccgcggtggt
     1321 ggcgtcacgc ctgctgcagc accacatcac ggagcagctg caggacatcg tggacatcct
     1381 gaagaactcc gccgtcctgc cccctacctg cctgggggag gagcctgaga acacgcccgc
     1441 caacagcgcc ccgggctccc ccagcacgcc ccccacacgc ttctttaccg agaagaagat
     1501 tccccatgag tgcctggtca tcggagcgct tgaaagtgca ttcaaggaaa tggacctaca
     1561 gatagaacga gagaggagtt catataatat atctggtggc tgcacggccc tcattgtgat
     1621 ttgccttttg gggaagctgt atgttgcaaa tgctggggat agcagggcca taatcatcag
     1681 aaatggagaa attatcccca tgtcttcaga atttaccccc gagacggagc gccagcgact
     1741 tcagtacctg gcattcatgc agcctcactt gctgggaaat gagttcacac atttggagtt
     1801 tccaaggaga gtacagagaa aggagcttgg aaagaagatg ctctacaggg actttaatat
     1861 gacaggctgg gcatacaaaa ccattgagga tgaggacttg aagttccccc ttatatatgg
     1921 agaaggcaag aaggcccggg taatggcaac tattggagtg accaggggac ttggggacca
     1981 tgacctgaag gtgcatgact ccaacatcta cattaaacca ttcctgtctt cagctccaga
     2041 ggtaagaatc tacgatcttt caaaatatga tcatggatca gatgatgtgc tgatcttggc
     2101 cactgatgga ctctgggacg ttttatcaaa tgaagaagta gcagaagcaa tcactcagtt
     2161 tcttcctaac tgtgatccag atgatcctca caggtacaca ctggcagctc aggacctggt
     2221 gatgcgtgcc cggggtgtgc tgaaggacag aggatggcgg atatctaatg accgactggg
     2281 ctcaggagac gacatttctg tatatgtcat tcctttaata catggaaaca agctgtcatg
     2341 agcggccgct tacccatacg atgtgccaga ttacgcctaa gtcgagtcta gagggcccgt
     2401 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2461 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2521 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2581 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     2641 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     2701 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2761 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2821 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2881 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtacct agaagttcct
     2941 attccgaagt tcctattctc tagaaagtat aggaacttcc ttggccaaaa agcctgaact
     3001 caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat
     3061 gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata
     3121 tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca
     3181 ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggagt tcagcgagag
     3241 cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac
     3301 cgaactgccc gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga
     3361 tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac
     3421 atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat
     3481 ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga
     3541 ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac
     3601 ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca
     3661 atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac
     3721 gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat
     3781 gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc
     3841 agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg
     3901 tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc
     3961 cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaatagc acgtactacg
     4021 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4081 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4141 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4201 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4261 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4321 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4381 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4441 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4501 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4561 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4621 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4681 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4741 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4801 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4861 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4921 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4981 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5041 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5101 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5161 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5221 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5281 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5341 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5401 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5461 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5521 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5581 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5641 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5701 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5761 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5821 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5881 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5941 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6001 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6061 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6121 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6181 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6241 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6301 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6361 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6421 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
