LOCUS       sequence-381314-        7835 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Express human Plekhm1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7835)
  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 7835)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_EGFP
FEATURES             Location/Qualifiers
     source          1..7835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3847..3856
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3853..3855,3856..3858,3859..4572)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3853..3855,3856..3858,3859..4569)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3898..3919)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4159..4178)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4506..4527
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4695..4816
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4732..4751)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4786..4805
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4823..5278)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4960..4981)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5172..5191
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5305..5409
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5434..5454)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5478..5688
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        5500..5696
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5547..5682
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5731..6525
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5731..6525
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5785..5804)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6395..6414
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6713..6732)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6757..6804
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7133..7721
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7622..7641
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagtg agtggagatg ctttcagtgg tggagaatgg actggacccc
      661 caggctgcca tcccggtcat caagaagaag ctggtgggat ccgtgaaggc cttgcagaag
      721 cagtacgtgt ccctggacac ggtggtcact agtgaagacg gagatgccaa caccatgtgc
      781 agcgccctgg aggccgtatt tatccatggc ctgcacgcca agcacatccg agctgaggcc
      841 ggaggaaaaa ggaagaaaag tgcccaccag aagcctctgc cccagcctgt cttctggccc
      901 ctcctgaaag ctgtcaccca caaacacatc atctcagagt tggagcacct gacgtttgtc
      961 aacacggatg tgggccgctg ccgggcatgg ctgcggctgg ccctgaacga tggcctgatg
     1021 gagtgctacc tgaagctgct gctgcaggag caggcccgct tgcatgagta ctaccagccc
     1081 accgccctgc tccgggatgc tgaggagggc gagttcctcc ttagcttcct gcagggcctc
     1141 acgtccttgt ccttcgaact ctcctacaag tctgccatct taaatgagtg gacgctcacc
     1201 ccattggccc tgtctgggct ttgcccgctt tctgagctgg accctctctc tacctctggt
     1261 gcagaactac agcggaagga atctctggat tccatttccc attcttcagg ctctgaagac
     1321 atcgaagtcc atcactcggg ccataagata cggaggaacc agaagctgac tgcctcctcc
     1381 ctcagcctgg acacggccag ttcatcccag ctgtcctgca gcctaaactc tgatagctgc
     1441 ttactccaag agaatggctc caagagtcca gaccattgcg aggagcccat gtcctgtgac
     1501 tcagacctgg gcacagcaaa tgctgaggac tcagaccggt ctctgcaaga ggtattgttg
     1561 gaattcagca aagcccaggt aaactctgtg ccaaccaacg gactgagcca agaaacagag
     1621 atccccacac cacaggcctc gctctccctc catggcctca acaccagcac atacctgcac
     1681 tgtgaggcac ctgcagagcc ccttcctgcc caggcagcct ctggaactca agatggtgtc
     1741 cacgtgcagg agccgcgtcc ccaggcgccc agccccctgg acttacagca gcctgtagag
     1801 agcacctcag gccagcagcc ttctagtact gtcagcgaga cagccagaga agtgggccaa
     1861 gggaatggcc tgcagaaggc ccaggctcat gacggagctg gtctgaagct ggtagtttcc
     1921 tcacccacca gtccgaaaaa caagagctgg atctcagagg atgacttcta ccggccttcc
     1981 cgggagcaac ccctggagag tgcttcagac cacccaatag cttcttacag ggggactcca
     2041 gggtcaaggc ctggtctcca caggcatttt tctcaagaac caagaaaaaa ctgctccctg
     2101 ggggcgttag accaagcgtg tgtaccttcc ccaggaagaa ggcaagccca ggcagcccca
     2161 tcccaggggc ataagagctt ccgggtggta caccggagac agatgggact gtccaaccca
     2221 ttccggggtc tcatgaagct gggcaccgtg gagcggcggg gggcaatggg catctggaag
     2281 gagctcttct gcgagctctc cccgctggag ttccgcctct acctgagcaa cgaggagcac
     2341 acctgtgtgg agaactgctc gctgcttcgc tgtgagtctg tggggccagc ccatagtgat
     2401 gggcgctttg agctggtctt ctctggcaag aagctggccc tgcgcgcctc ctcccaggac
     2461 gaagctgagg actggctgga ccgggtgcgg gaggccctgc agaaggtccg gcctcagcag
     2521 gaggatgagt gggtgaacgt gcagtaccca gaccagcctg aggaaccccc cgaggcgccc
     2581 cagggctgcc tctctccctc agacctgctc tcggagcccg cggccctcca gggcacacag
     2641 tttgactggt cgtccgccca ggttccagag ccagatgcca tcaaggagtc cctgctgtac
     2701 ttgtacatgg acaggacctg gatgccctat atattttctc tgtccttgga ggctctgaaa
     2761 tgtttccgca tcaggaacaa tgagaagatg ctgagtgaca gccacggcgt ggagaccatc
     2821 cgggacatcc tgccagacac cagccttggg ggcccatcct tcttcaaaat catcacggcc
     2881 aaggctgtcc tgaagctgca ggccggaaac gccgaggaag ccgccctgtg gagggatctg
     2941 gtccgcaaag tcctggcatc ctacttggag acagccgagg aggcggtgac cctgggcggg
     3001 agcctggatg aaaactgtca ggaggtgctg aaatttgcca cccgggagaa tggcttcctg
     3061 ctgcagtacc tggtggctat ccccatggag aaaggccttg actcccaagg ctgcttctgc
     3121 gcaggctgct cccggcagat cggcttctcc tttgtacgac ccaagctctg tgccttctct
     3181 ggcctctatt actgtgacat ctgccaccaa gacgatgcct cagtgattcc ggccaggatc
     3241 atccacaact gggacctcac caagcgcccg atctgcaggc aggccctgaa gtttctgaca
     3301 cagatccggg cccagcccct catcaacctg cagatggtga acgcgtctct gtacgagcat
     3361 gtggagcgga tgcacctcat tgggaggaga cgggagcagc tgaagctcct gggggattac
     3421 ctgggcctgt gccggagtgg cgccctgaag gagctcagca agaggctcaa ccacaggaat
     3481 tatctcttgg aatctccgca taggttcagt gttgctgacc tccaacagat cgcagacggg
     3541 gtgtatgaag gattcctcaa ggccctgatt gaatttgcct cccagcatgt ctaccactgc
     3601 gacctgtgca cccagcgcgg cttcatctgc cagatctgcc agcaccacga catcatcttc
     3661 ccctttgagt ttgacaccac agtcaggtgt gccgagtgca agaccgtctt ccaccagagc
     3721 tgccaggctg tggtgaagaa gggctgcccc cgctgtgccc gccggcgcaa gtaccaggaa
     3781 cagaacattt tcgccaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatcca
     3841 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     3901 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     3961 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     4021 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     4081 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     4141 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     4201 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     4261 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     4321 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     4381 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     4441 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     4501 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     4561 ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     4621 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     4681 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4741 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4801 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     4861 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     4921 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     4981 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     5041 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     5101 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     5161 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     5221 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     5281 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5341 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5401 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     5461 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5521 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5581 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     5641 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     5701 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     5761 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     5821 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     5881 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     5941 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     6001 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     6061 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     6121 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     6181 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     6241 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     6301 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     6361 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     6421 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     6481 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     6541 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     6601 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     6661 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     6721 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     6781 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     6841 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     6901 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     6961 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     7021 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     7081 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     7141 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     7201 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     7261 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7321 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7381 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7441 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7501 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7561 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7621 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7681 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7741 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     7801 ctgattctgt ggataaccgt attaccgcca tgcat
//
