LOCUS       sequence-413041-        6506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of His SUMO Rab10(Q68L).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6506)
  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 6506)
  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: pET15b+His SUMO Rab10(Q68L)
FEATURES             Location/Qualifiers
     source          1..6506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..15,16..32,33..38)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(213..260)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     256..274
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(1170..1187)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1200..1222
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1237..1261
                     /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)."
     primer_bind     complement(1261..1280)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1262..1280)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1364..1383)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1589..1666
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1667..2749
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1686..1705)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4061..4252
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4246..4268)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4354..4493
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4409..4428
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4508..4525)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4679..5267)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4759..4778)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5438..6229,6230..6298))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6061..6080
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6299..6403)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6471..6489
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gtctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
      241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
      301 cgggctttgt tagcagccgg atctttacgc ggccgtcaac tgttgggctc ttttacaggg
      361 gtctttcgaa ggatatcttc agctaacgtg aggaacgcct tttcgatgtt tatatttgct
      421 tttgcactag tctcaaaaaa cctaatacca tgctcccttg caatctgttc tccttttcct
      481 ttaggtacaa ctcttttgtc gtccatatca cacttgtttc ctagtaacat tctttccaca
      541 tcttcattgg catgctcatc tatgtttcta agccatttgc tgatgttttc aaaactttta
      601 ccattggtga tgtcatatac tagcatgata cccattgcgc ctctgtagta ggaggttgtg
      661 atggtgtgaa atcgctccag gcctgctgta tcccatatct gtagcttgat cttctttcct
      721 tgtaattcaa ctgttttgat cttgaagtct attcctatgg tggaaataaa ggtagtattg
      781 aaggcatcat ccgaaaaacg aaaaaggacg caggtcttcc ccactccgga atccccgatc
      841 aggagcagct tgaaaagcag gtcgtacgtc ttcttcgcca ttccccccgt ttgttcctga
      901 taaacttcaa tcacatcttc ttcctccatt cccagttctt ttggagtatg attatcagca
      961 attctctgac cctcaaagag aaacctgagt gagttcattg gaacaccctg tctttgacag
     1021 tatgattctt tgagtttctt gagatgtgtt gtcattttca ctttgaagtg aatctcactg
     1081 ctatcctgtc caatgacttt gagtttaata tattcacctt ccttcttatc ccccaagtcc
     1141 tcagttgaag gttttgcctc ctggtcagaa tggtgatggt gatgatgacc catggtatat
     1201 ctccttctta aagttaaaca aaattatttc tagaggggaa ttgttatccg ctcacaattc
     1261 ccctatagtg agtcgtatta atttcgcggg atcgagatct cgatcctcta cgccggacgc
     1321 atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc
     1381 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt
     1441 atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc
     1501 cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag
     1561 tcgcataagg gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa cctttcgcgg
     1621 tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac
     1681 gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa
     1741 ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct
     1801 gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg
     1861 cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc
     1921 tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga
     1981 agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa
     2041 ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc
     2101 gttatttctt gatgtctctg accagacacc catcaacagt attattttct cccatgaaga
     2161 cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt
     2221 agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct
     2281 cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg
     2341 ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc
     2401 caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg
     2461 tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc
     2521 gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct
     2581 gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa
     2641 aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     2701 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     2761 ttaatgtaag ttagctcact cattaggcac cgggatctcg accgatgccc ttgagagcct
     2821 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
     2881 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccgga agcgctctgg gtcattttcg
     2941 gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa
     3001 tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga
     3061 agcaggccat tatcgccggc atggcggccg acgcgctggg ctacgtcttg ctggcgttcg
     3121 cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc ggcatcggga
     3181 tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag ggacagcttc
     3241 aaggatcgct cgcggctctt accagcctaa cttcgatcat tggaccgctg atcgtcacgg
     3301 cgatttatgc cgcctcggcg agcacatgga acgggttggc atggattgta ggcgccgccc
     3361 tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct
     3421 gaatggaagc cggcggcacc tcgctaacgg attcaccact ccaagaattg gagccaatca
     3481 attcttgcgg agaactgtga atgcgcaaac caacccttgg cagaacatat ccatcgcgtc
     3541 cgccatctcc agcagccgca cgcggcgcat ctcgggcagc gttgggtcct ggccacgggt
     3601 gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg
     3661 ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc
     3721 tgcgacctga gcaacaacat gaatggtctt cggtttacgt gtttcgtaaa gtctggaaac
     3781 gcggaagtca gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct
     3841 accctgtgga acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt
     3901 tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg
     3961 ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat
     4021 taccccaatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac
     4081 cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa
     4141 cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga
     4201 gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     4261 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     4321 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     4381 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
     4441 cattgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
     4501 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4561 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4621 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4681 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4741 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4801 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4861 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4921 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4981 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5041 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5101 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5161 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5221 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5281 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5341 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5401 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5461 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5521 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5581 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5641 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5701 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca
     5761 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5821 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5881 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5941 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6001 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg
     6061 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6121 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6181 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6241 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6301 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6361 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6421 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     6481 tcacgaggcc ctttcgtctt caagaa
//
