LOCUS       sequence-337052-        7514 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transfer plasmid for lentiviral vector production with miR for Hs 
            ATP10B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7514)
  AUTHORS   Martin S, Smolders S, Van den Haute C, Heeman B, van Veen S, 
            Crosiers D, Beletchi I, Verstraeten A, Gossye H, Gelders G, Pals P, 
            Hamouda NN, Engelborghs S, Martin JJ, Eggermont J, De Deyn PP, Cras 
            P, Baekelandt V, Vangheluwe P, Van Broeckhoven C
  TITLE     Mutated ATP10B increases Parkinson's disease risk by compromising 
            lysosomal glucosylceramide export.
  JOURNAL   Acta Neuropathol. 2020 Jun;139(6):1001-1024. doi: 
            10.1007/s00401-020-02145-7. Epub 2020 Mar 14.
  PUBMED    32172343
REFERENCE   2  (bases 1 to 7514)
  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"; doi: 
            "10.1007/s00401-020-02145-7"; journalName: "Acta Neuropathol"; date:
            "2020-06"; volume: "139"; issue: "6"; pages: "1001-1024"
COMMENT     Sequence Label: miR3 HsATP10B
FEATURES             Location/Qualifiers
     source          1..7514
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(529..1320,1321..1389))
                     /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     1152..1171
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1390..1494)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1520..1975)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1657..1678)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1869..1888
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2102..2124
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2116..2133
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2117..2133
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2140..2159
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2140..2158
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        2273..2652
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2653..2856
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2807..2827
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        4546..4953
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             4977..5375
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     ncRNA           5440..5454
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     misc_feature    5547..6135
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5600..5620)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6018..6029)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(6843..6861)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6882..6898)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6882..6898)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6895..6917)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6906..6922
                     /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(6930..6936,6937..6954,6955..6960))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6975..6996
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7113..7130)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(7284..7514,1..358))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7364..7383)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
       61 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
      121 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
      181 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
      241 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
      301 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
      361 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
      421 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
      481 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
      541 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
      601 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
      661 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
      721 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
      781 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
      841 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
      901 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
      961 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     1021 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     1081 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     1141 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     1201 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     1261 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     1321 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     1381 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     1441 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     1501 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     1561 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     1621 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     1681 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     1741 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     1801 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     1861 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     1921 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     1981 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     2041 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     2101 tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc
     2161 gaattggagc tccaccgcgg tggcggccgc tctagaattc cattgcatac gttgtatcca
     2221 tatcataata tgtacattta tattggctca tgtccaacat taccgccatg ttgacattga
     2281 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     2341 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     2401 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     2461 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     2521 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     2581 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     2641 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     2701 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     2761 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     2821 aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg cagtcgctct
     2881 gcggagaggc tggcagattg agccctggga ggttctctcc agcactagca ggtagagcct
     2941 gggtgttccc tgctagactc tcaccagcac ttggccagtg ctgggcagag tggctccacg
     3001 cttgcttgct taaagacctc ttcaataaag ctgccatttt agaagtaagc cagtgtgtgt
     3061 tcccatctct cctagtcgcc gcctggtcaa ctcggtactc ggtaataaga agaccctggt
     3121 ctgttaggac cctttctgct ttgagaaacc gaagcaggaa aatccctagc agattggcgc
     3181 ccgaacaggg acttgaagga gagtgagaga ctcctgagta cggctgagtg aaggcagtaa
     3241 gggcggcagg aaccaaccac gacggagtgc tcctataaag gcgcgggtcg gtaccagacg
     3301 gcgtgaggag cgggagagga ggaggcctcc ggttgcaggt aagtgcaaca caaaaaagaa
     3361 atagctgtct tgttatccag gaagggataa taagatagag tgggagatgg gcgcgagaaa
     3421 ctccgtcttg tcagggaaga aagcagatga attgaaaaaa ttattaatcg catgaatttt
     3481 aaaagaaggg gaggaatagg ggatatgact ccagcagaaa gattaattaa catgatcact
     3541 acagaacaag aaatacaatt tcaacaatca aaaaactcaa aatttaaaaa ttttcgggtc
     3601 tattacagag ctcacgcgtg attggagttg ggagattata aattagtaga gatcactccg
     3661 attggcttgg cccccacaga tgtgaagagg tacactactg gtggcacctc aagaaataaa
     3721 agaggggtct ttgtgctagg gttcttgggt tttctcgcaa cggcaggttc tgcaatgggc
     3781 gcggcgtcgt tgacgctgac cgctcagtcc cggactttat tggctgggat agtgcagcaa
     3841 cagcaacagc tgttggacgt ggtcaagaga caacaagaat tgttgcgact gaccgtctgg
     3901 ggaacaaaga acctccagac tagggtcact gccatcgaga agtacttaaa ggaccaggcg
     3961 cagctaaatg cttggggatg tgcgtttaga caagtctgcc acactactgt accatggcca
     4021 aatgcaagtc taacaccaga ctggaacaat gatacttggc aagagtggga gcgaaaggtt
     4081 gacttcttgg aggaaaatat aacagccctc ctagaagagg cacaaattca acaagagaag
     4141 aacatgtatg aattacaaaa gttgaatagc tgggatgtgt ttggcaattg gtttgacctt
     4201 gcttcttgga taaagtatat acaatatgga atttatgtag ttgtaggagt aatactgtta
     4261 agaatagtga tctatatagt acaaatgcta gctaagttaa ggcaggggta taggccagtg
     4321 ttctcttccc caccctctta tttccagtag actcataccc aacaggaccc ggcactgcca
     4381 accagagaag gcaaagaagg acggtggaga aggcggtgga tctgctagca ctagttatta
     4441 atcgtacgat taactgcagc cccgataaaa taaaagattt tatttagtct ccagaaaaag
     4501 gggggaatga aagaccccac ctgtaggttt ggcaagctag ctgcagtaac gccattttgc
     4561 aaggcatgga aaaataccaa accaagaata gagaagttca gatcaagggc gggtacatga
     4621 aaatagctaa cgttgggcca aacaggatat ctgcggtgag cagtttcggc cccggcccgg
     4681 ggccaagaac agatggtcac cgcagtttcg gccccggccc gaggccaaga acagatggtc
     4741 cccagatatg gcccaaccct cagcagtttc ttaagaccca tcagatgttt ccaggctccc
     4801 ccaaggacct gaaatgaccc tgcgccttat ttgaattaac caatcagcct gcttctcgct
     4861 tctgttcgcg cgcttctgct tcccgagctc tataaaagag ctcacaaccc ctcactcggc
     4921 gcgccagtcc tccgacagac tgagtcgccc gggggaccgg gagcagatct accggtatgg
     4981 ccaagccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca
     5041 gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca
     5101 tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc
     5161 tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg
     5221 agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc
     5281 ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg
     5341 aattgctgcc ctctggttat gtgtgggagg gctaactcga gatccaagaa ggtatattgc
     5401 tgttgacagt gagcgtcttt gccatcaccc gctttaatag tgaagccaca gatgtattaa
     5461 agcgggtgat ggcaaagttg cctactgcct cggacttcaa gggctacgat gtcgacttgg
     5521 atccatcgat ggatctaatt cccgataatc aacctctgga ttacaaaatt tgtgaaagat
     5581 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     5641 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     5701 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     5761 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     5821 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     5881 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     5941 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     6001 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     6061 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     6121 gggccgcctc cccgcatcgt cgagttttat aaaagaaaag gggggactgg aagggattta
     6181 ttacagtgca agaagacata gaatcttaga catgtactta gaaaaggaag aaggcatcat
     6241 accagattgg caggattaca cctcaggacc aggaattaga tacccaaaga catttggctg
     6301 gctatggaaa ttagtccctg taaatgtatc agatgaggca caggaggatg agaggcatta
     6361 tttaatgcag ccagctcaaa cttccaagtg ggatgaccct tggggagagg ttctagcgtg
     6421 gaagtttgat ccaactctag cctacactta tgaggcatat gctagatacc cagaagagtt
     6481 ggaagcaagt caggcctgtc agaactgcat ttcgctctgt attcagtcgc tctgcggaga
     6541 ggctggcaga ttgagccctg ggaggttctc tccagcacta gcaggtagag cctgggtgtt
     6601 ccctgctaga ctctcaccag cacttggcca gtgctgggca gagtggctcc acgcttgctt
     6661 gcttaaagac ctcttcaata aagctgccat tttagaagta agccagtgtg tgttcccatc
     6721 tctcctagtc gccgcctggt caactcggta ctcggtaata agaagaccct ggtctgttag
     6781 gaccctttct gctttgagaa accgaagcag gaaaatccct agcatggtac cagcttttgt
     6841 tccctttagt gagggttaat ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg
     6901 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6961 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     7021 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     7081 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     7141 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     7201 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     7261 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     7321 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7381 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7441 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     7501 agttcggtgt aggt
//
