LOCUS       sequence-412902-        6355 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of HA-Delta-37, 115-133, 204-217 PPM1H.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6355)
  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 6355)
  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-37, 115-133, 204-217 PPM1H
FEATURES             Location/Qualifiers
     source          1..6355
                     /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             2240..2266
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2307..2324)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2313..2537
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2670..2689)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2821..2868
                     /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             2876..3898
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4028..4149
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4065..4084)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4119..4138
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4198..4214)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4198..4214)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4211..4233)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4222..4238
                     /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(4246..4252,4253..4270,4271..4276))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4291..4312
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4429..4446)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4600..5188)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4680..4699)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5359..6150,6151..6219))
                     /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     5982..6001
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6220..6324)
                     /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 gatcccgttt
      901 cccctacggg cggccagagt tcctggggct gtctcaggac gaggtggagt gcagcgccga
      961 ccacatcgcc cgccccatcc tcatcctcaa ggagactcgg cggctgccct gggccactgg
     1021 ctacgcagag gttatcaatg ccgggaagag cacacacaat gaagaccaag ccagctgtga
     1081 ggtgctcact gtgaagaaga aggcaggggc cgtgacctca accccaaagg agaactcgga
     1141 atccgagggt gtttcctgcc actattggtc gctgtttgac gggcacgcgg ggtccggggc
     1201 cgcggtggtg gcgtcacgcc tgctgcagca ccacatcacg gagcagctgc aggacatcgt
     1261 ggacatcctg aagaactccg ccgtcctgcc ccctacctgc ctgggggagg agcctgagaa
     1321 cacgcccgcc aacagcgccc cgggctcccc cagcacgccc cccacacgct tctttaccga
     1381 gaagaagatt ccccatgagt gcctggtcat cggagcgctt gaaagtgcat tcaaggaaat
     1441 ggacctacag atagaacgag agaggagttc atataatata tctggtggct gcacggccct
     1501 cattgtgatt tgccttttgg ggaagctgta tgttgcaaat gctggggata gcagggccat
     1561 aatcatcaga aatggagaaa ttatccccat gtcttcagaa tttacccccg agacggagcg
     1621 ccagcgactt cagtacctgg cattcatgca gcctcacttg ctgggaaatg agttcacaca
     1681 tttggagttt ccaaggagag tacagagaaa ggagcttgga aagaagatgc tctacaggga
     1741 ctttaatatg acaggctggg catacaaaac cattgaggat gaggacttga agttccccct
     1801 tatatatgga gaaggcaaga aggcccgggt aatggcaact attggagtga ccaggggact
     1861 tggggaccat gacctgaagg tgcatgactc caacatctac attaaaccat tcctgtcttc
     1921 agctccagag gtaagaatct acgatctttc aaaatatgat catggatcag atgatgtgct
     1981 gatcttggcc actgatggac tctgggacgt tttatcaaat gaagaagtag cagaagcaat
     2041 cactcagttt cttcctaact gtgatccaga tgatcctcac aggtacacac tggcagctca
     2101 ggacctggtg atgcgtgccc ggggtgtgct gaaggacaga ggatggcgga tatctaatga
     2161 ccgactgggc tcaggagacg acatttctgt atatgtcatt cctttaatac atggaaacaa
     2221 gctgtcatga gcggccgctt acccatacga tgtgccagat tacgcctaag tcgagtctag
     2281 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2341 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2401 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2461 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     2521 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     2581 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2641 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2701 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2761 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtaccta
     2821 gaagttccta ttccgaagtt cctattctct agaaagtata ggaacttcct tggccaaaaa
     2881 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     2941 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     3001 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     3061 ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggagtt
     3121 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     3181 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc
     3241 tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     3301 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     3361 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     3421 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     3481 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     3541 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     3601 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg
     3661 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     3721 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     3781 tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga
     3841 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagca
     3901 cgtactacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     3961 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4021 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4081 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4141 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4201 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4261 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4321 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4381 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4441 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4501 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4561 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4621 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4681 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4741 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4801 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4861 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4921 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4981 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5041 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5101 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5161 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5221 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5281 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5341 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5401 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5461 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5521 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5581 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5641 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5701 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5761 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5821 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5881 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5941 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6001 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6061 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6121 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6181 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6241 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6301 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
