LOCUS       sequence-412896-        6463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of HA-Delta-34 PPM1H.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6463)
  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 6463)
  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-34 PPM1H
FEATURES             Location/Qualifiers
     source          1..6463
                     /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             2348..2374
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2415..2432)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2421..2645
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2778..2797)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2929..2976
                     /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             2984..4006
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4136..4257
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4173..4192)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4227..4246
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4306..4322)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4306..4322)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4319..4341)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4330..4346
                     /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(4354..4360,4361..4378,4379..4384))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4399..4420
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4537..4554)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4708..5296)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4788..4807)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5467..6258,6259..6327))
                     /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     6090..6109
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6328..6432)
                     /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 gatccctgcc
      901 cctgcgtttc ccctacgggc ggccagagtt cctggggctg tctcaggacg aggtggagtg
      961 cagcgccgac cacatcgccc gccccatcct catcctcaag gagactcggc ggctgccctg
     1021 ggccactggc tacgcagagg ttatcaatgc cgggaagagc acacacaatg aagaccaagc
     1081 cagctgtgag gtgctcactg tgaagaagaa ggcaggggcc gtgacctcaa ccccaaacag
     1141 gaactcatcc aagagacggt cctcccttcc caatggggaa gggctgcagc tgaaggagaa
     1201 ctcggaatcc gagggtgttt cctgccacta ttggtcgctg tttgacgggc acgcggggtc
     1261 cggggccgcg gtggtggcgt cacgcctgct gcagcaccac atcacggagc agctgcagga
     1321 catcgtggac atcctgaaga actccgccgt cctgccccct acctgcctgg gggaggagcc
     1381 tgagaacacg cccgccaaca gccggactct gacccgggca gcctccctgc gcggaggggt
     1441 gggggccccg ggctccccca gcacgccccc cacacgcttc tttaccgaga agaagattcc
     1501 ccatgagtgc ctggtcatcg gagcgcttga aagtgcattc aaggaaatgg acctacagat
     1561 agaacgagag aggagttcat ataatatatc tggtggctgc acggccctca ttgtgatttg
     1621 ccttttgggg aagctgtatg ttgcaaatgc tggggatagc agggccataa tcatcagaaa
     1681 tggagaaatt atccccatgt cttcagaatt tacccccgag acggagcgcc agcgacttca
     1741 gtacctggca ttcatgcagc ctcacttgct gggaaatgag ttcacacatt tggagtttcc
     1801 aaggagagta cagagaaagg agcttggaaa gaagatgctc tacagggact ttaatatgac
     1861 aggctgggca tacaaaacca ttgaggatga ggacttgaag ttccccctta tatatggaga
     1921 aggcaagaag gcccgggtaa tggcaactat tggagtgacc aggggacttg gggaccatga
     1981 cctgaaggtg catgactcca acatctacat taaaccattc ctgtcttcag ctccagaggt
     2041 aagaatctac gatctttcaa aatatgatca tggatcagat gatgtgctga tcttggccac
     2101 tgatggactc tgggacgttt tatcaaatga agaagtagca gaagcaatca ctcagtttct
     2161 tcctaactgt gatccagatg atcctcacag gtacacactg gcagctcagg acctggtgat
     2221 gcgtgcccgg ggtgtgctga aggacagagg atggcggata tctaatgacc gactgggctc
     2281 aggagacgac atttctgtat atgtcattcc tttaatacat ggaaacaagc tgtcatgagc
     2341 ggccgcttac ccatacgatg tgccagatta cgcctaagtc gagtctagag ggcccgttta
     2401 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2461 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2521 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2581 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     2641 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     2701 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2761 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2821 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2881 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtacctaga agttcctatt
     2941 ccgaagttcc tattctctag aaagtatagg aacttccttg gccaaaaagc ctgaactcac
     3001 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     3061 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
     3121 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
     3181 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggagttca gcgagagcct
     3241 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
     3301 actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct
     3361 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     3421 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     3481 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     3541 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     3601 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     3661 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     3721 ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct
     3781 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     3841 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     3901 acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga
     3961 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagcacg tactacgaga
     4021 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4081 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4141 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4201 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4261 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     4321 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4381 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4441 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4501 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4561 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4621 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4681 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4741 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4801 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4861 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4921 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4981 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5041 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5101 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5161 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5221 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5281 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5341 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5401 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5461 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5521 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5581 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5641 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5701 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5761 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5821 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5881 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5941 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6001 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6061 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6121 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6181 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6241 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6301 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6361 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6421 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
