LOCUS       sequence-416695-        6149 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of myc-tagged human Rab10 with C-terminal mito 
            signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6149)
  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 6149)
  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: pcDNA 3.1 myc-Rab10-mito
FEATURES             Location/Qualifiers
     source          1..6149
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        190..569
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        570..773
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     724..744
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     818..837
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        818..836
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             859..888
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(1657..1675)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1658..1675)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1695..1712)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1701..1925
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1971..2399
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2058..2077)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2268..2289
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2408..2428)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2413..2742
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2593..2728
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2655..2674
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2809..3603
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2863..2882)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3473..3492
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3777..3898
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3814..3833)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3868..3887
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3947..3963)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3947..3963)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3960..3982)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3971..3987
                     /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(3995..4001,4002..4019,4020..4025))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4040..4061
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4178..4195)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4349..4937)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4429..4448)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5108..5899,5900..5968))
                     /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     5731..5750
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5969..6073)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(6148..6149,1..18))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
       61 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
      121 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
      181 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      241 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      301 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      361 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      421 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      481 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      541 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      601 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      661 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      721 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      781 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      841 cccaagcttg gtaccatgga acaaaaactc atctcagaag aggatctgaa tttatcaatt
      901 accggtggat ccatggcgaa gaagacgtac gacctgcttt tcaagctgct cctgatcggg
      961 gattccggag tggggaagac ctgcgtcctt tttcgttttt cggatgatgc cttcaatact
     1021 acctttattt ccaccatagg aatagacttc aagatcaaaa cagttgaatt acaaggaaag
     1081 aagatcaagc tacagatatg ggatacagca ggccaggagc gatttcacac catcacaacc
     1141 tcctactaca gaggcgcaat gggtatcatg ctagtatatg acatcaccaa tggtaaaagt
     1201 tttgaaaaca tcagcaaatg gcttagaaac atagatgagc atgccaatga agatgtggaa
     1261 agaatgttac taggaaacaa gtgtgatatg gacgacaaaa gagttgtacc taaaggaaaa
     1321 ggagaacaga ttgcaaggga gcatggtatt aggttttttg agactagtgc aaaagcaaat
     1381 ataaacatcg aaaaggcgtt cctcacgtta gctgaagata tccttcgaaa gacccctgta
     1441 aaagagccca acagtgaaaa tgtagatatc agcagtggag gaggcgtgac aggctggaag
     1501 agcaaaggtg ctggcgccgg tgcggccgcc ttctgggaaa ggaacctgcc ctctgtttct
     1561 ggcctgctga agatcattgg attttccaca tcagtaactg ccctggggtt tgtgctgtac
     1621 aaatacaagc tcctgccacg gtcttgaggg ccctattcta tagtgtcacc taaatgctag
     1681 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     1741 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     1801 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     1861 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     1921 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     1981 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2041 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2101 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2161 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2221 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2281 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2341 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     2401 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     2461 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     2521 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     2581 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2641 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     2701 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     2761 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     2821 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     2881 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     2941 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3001 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3061 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3121 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3181 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3241 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3301 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     3361 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3421 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3481 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     3541 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     3601 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     3661 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     3721 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     3781 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3841 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3901 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     3961 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4021 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4081 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4141 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4201 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4261 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4321 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4381 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4441 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4501 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4561 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4621 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4681 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4741 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4801 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4861 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4921 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4981 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5041 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5101 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5161 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5221 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5281 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5341 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5401 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5461 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5521 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5581 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5641 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5701 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5761 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5821 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5881 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5941 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6001 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6061 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     6121 tcccgatccc ctatggtgca ctctcagta
//
