LOCUS       sequence-140090-        8551 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human Plekhm1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8551)
  AUTHORS   Van Wesenbeeck L, Odgren PR, Coxon FP, Frattini A, Moens P, Perdu B,
            MacKay CA, Van Hul E, Timmermans JP, Vanhoenacker F, Jacobs R, 
            Peruzzi B, Teti A, Helfrich MH, Rogers MJ, Villa A, Van Hul W
  TITLE     Involvement of PLEKHM1 in osteoclastic vesicular transport and 
            osteopetrosis in incisors absent rats and humans.
  JOURNAL   J Clin Invest. 2007 Apr;117(4):919-30.
  PUBMED    17404618
REFERENCE   2  (bases 1 to 8551)
  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: "J Clin 
            Invest."; date: "2007-04"; volume: "117(4)"; pages: "919-30"
COMMENT     Sequence Label: h-Plekhm1_FLAG-pcDNA 3.1
FEATURES             Location/Qualifiers
     source          1..8551
                     /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"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4072..4095
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4145..4162)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4151..4375
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4421..4849
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4508..4527)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4718..4739
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4858..4878)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4863..5192
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5043..5178
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5105..5124
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5259..6053
                     /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(5313..5332)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5923..5942
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6227..6348
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6264..6283)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6318..6337
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6397..6413)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6397..6413)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6410..6432)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6421..6437
                     /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(6445..6451,6452..6469,6470..6475))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6490..6511
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6628..6645)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6799..7384)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6879..6898)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7555..8346,8347..8415))
                     /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     8178..8197
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8416..8520)
                     /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 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctttgagtg
      901 gagatgcttt cagtggtgga gaatggactg gacccccagg ctgccatccc ggtcatcaag
      961 aagaagctgg tgggatccgt gaaggccttg cagaagcagt acgtgtccct ggacacggtg
     1021 gtcactagtg aagacggaga tgccaacacc atgtgcagcg ccctggaggc cgtatttatc
     1081 catggcctgc acgccaagca catccgagct gaggccggag gaaaaaggaa gaaaagtgcc
     1141 caccagaagc ctctgcccca gcctgtcttc tggcccctcc tgaaagctgt cacccacaaa
     1201 cacatcatct cagagttgga gcacctgacg tttgtcaaca cggatgtggg ccgctgccgg
     1261 gcatggctgc ggctggccct gaacgatggc ctgatggagt gctacctgaa gctgctgctg
     1321 caggagcagg cccgcttgca tgagtactac cagcccaccg ccctgctccg ggatgctgag
     1381 gagggcgagt tcctccttag cttcctgcag ggcctcacgt ccttgtcctt cgaactctcc
     1441 tacaagtctg ccatcttaaa tgagtggacg ctcaccccat tggccctgtc tgggctttgc
     1501 ccgctttctg agctggaccc tctctctacc tctggtgcag aactacagcg gaaggaatct
     1561 ctggattcca tttcccattc ttcaggctct gaagacatcg aagtccatca ctcgggccat
     1621 aagatacgga ggaaccagaa gctgactgcc tcctccctca gcctggacac ggccagttca
     1681 tcccagctgt cctgcagcct aaactctgat agttgcttac tccaagagaa tggctccaag
     1741 agtccagacc attgcgagga gcccatgtcc tgtgactcag acctgggcac agcaaatgct
     1801 gaggactcag accggtctct gcaagaggta ttgttggaat tcagcaaagc ccaggtaaac
     1861 tctgtgccaa ccaacggact gagccaagaa acagagatcc ccacaccaca ggcctcgctc
     1921 tccctccatg gcctcaacac cagcacatac ctgcactgtg aggcacctgc agagcccctt
     1981 cctgcccagg cagcctctgg aactcaagat ggtgtccacg tgcaggagcc gcgtccccag
     2041 gcgcccagcc ccctggactt acagcagcct gtagagagca cctcaggcca gcagccttct
     2101 agtactgtca gcgagacagc cagagaagtg ggccaaggga atggcctgca gaaggcccag
     2161 gctcatgacg gagctggtct gaagctggta gtttcctcac ccaccagtcc gaaaaacaag
     2221 agctggatct cagaggatga cttctaccgg ccttcccggg agcaacccct ggagagtgct
     2281 tcagaccacc caatagcttc ttacaggggg actccagggt caaggcctgg tctccacagg
     2341 catttttctc aagaaccaag aaaaaactgc tccctggggg cgttagacca agcgtgtgta
     2401 ccttccccag gaagaaggca agcccaggca gccccatccc aggggcataa gagcttccgg
     2461 gtggtacacc ggagacagat gggactgtcc aacccattcc ggggtctcat gaagctgggc
     2521 accgtggagc ggcggggggc aatgggcatc tggaaggagc tcttctgcga gctctccccg
     2581 ctggagttcc gcctctacct gagcaacgag gagcacacct gtgtggagaa ctgctcgctg
     2641 cttcgctgtg agtctgtggg gccagcccat agtgatgggc gctttgagct ggtcttctct
     2701 ggcaagaagc tggccctgcg cgcctcctcc caggacgaag ctgaggactg gctggaccgg
     2761 gtgcgggagg ccctgcagaa ggtccggcct cagcaggagg atgagtgggt gaacgtgcag
     2821 tacccagacc agcctgagga accccccgag gcgccccagg gctgcctctc tccctcagac
     2881 ctgctctcgg agcccgcggc cctccagggc acacagtttg actggtcgtc cgcccaggtt
     2941 ccagagccag atgccatcaa ggagtccctg ctgtacttgt acatggacag gacctggatg
     3001 ccctatatat tttctctgtc cttggaggct ctgaaatgtt tccgcatcag gaacaatgag
     3061 aagatgctga gtgacagcca cggcgtggag accatccggg acatcctgcc agacaccagc
     3121 cttgggggcc catccttctt caaaatcatc acggccaagg ctgtcctgaa gctgcaggcc
     3181 ggaaacgccg aggaagccgc cctgtggagg gatctggtcc gcaaagtcct ggcatcctac
     3241 ttggagacag ccgaggaggc ggtgaccctg ggcgggagcc tggatgaaaa ctgtcaggag
     3301 gtgctgaaat ttgccacccg ggagaatggc ttcctgctgc agtacctggt ggctatcccc
     3361 atggagaaag gccttgactc ccaaggctgc ttctgcgcag gctgctcccg gcagatcggc
     3421 ttctcctttg tacgacccaa gctctgtgcc ttctctggcc tctattactg tgacatctgc
     3481 caccaagacg atgcctcagt gattccggcc aggatcatcc acaactggga cctcaccaag
     3541 cgcccgatct gcaggcaggc cctgaagttt ctgacacaga tccgggccca gcccctcatc
     3601 aacctgcaga tggtgaacgc gtctctgtac gagcatgtgg agcggatgca cctcattggg
     3661 aggagacggg agcagctgaa gctcctgggg gattacctgg gcctgtgccg gagtggcgcc
     3721 ctgaaggagc tcagcaagag gctcaaccac aggaattatc tcttggaatc tccgcatagg
     3781 ttcagtgttg ctgacctcca acagatcgca gacggggtgt atgaaggatt cctcaaggcc
     3841 ctgattgaat ttgcctccca gcatgtctac cactgcgacc tgtgcaccca gcgcggcttc
     3901 atctgccaga tctgccagca ccacgacatc atcttcccct ttgagtttga caccacagtc
     3961 aggtgtgccg agtgcaagac cgtcttccac cagagctgcc aggctgtggt gaagaagggc
     4021 tgcccccgct gtgcccgccg gcgcaagtac caggaacaga acattttcgc cgattacaag
     4081 gatgacgacg ataagatctg actcgagcat gcatctagag ggcccgttta aacccgctga
     4141 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     4201 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     4261 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     4321 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     4381 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     4441 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4501 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4561 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4621 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4681 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4741 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4801 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     4861 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4921 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4981 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     5041 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5101 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     5161 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     5221 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     5281 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     5341 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5401 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     5461 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5521 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5581 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5641 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5701 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5761 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     5821 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5881 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5941 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     6001 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     6061 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6121 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6181 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     6241 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6301 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     6361 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6421 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6481 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6541 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6601 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6661 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6721 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6781 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6841 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6901 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6961 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     7021 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7081 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7141 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7201 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     7261 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7321 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7381 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7441 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7501 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7561 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7621 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7681 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7741 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7801 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7861 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7921 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7981 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     8041 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     8101 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     8161 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8221 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8281 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8341 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8401 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8461 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     8521 cacatttccc cgaaaagtgc cacctgacgt c
//
