LOCUS       sequence-337051-        7514 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transfer plasmid for lentiviral vector production with miR for Mm 
            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: miR7 MmATP10B
FEATURES             Location/Qualifiers
     source          1..7514
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        756..1135
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1136..1339
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1290..1310
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3029..3436
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             3460..3858
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     ncRNA           3923..3937
                     /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    4030..4618
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4083..4103)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4501..4512)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(5326..5344)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5365..5381)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5365..5381)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5378..5400)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5389..5405
                     /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(5413..5419,5420..5437,5438..5443))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5458..5479
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5596..5613)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5767..6355)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5847..5866)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6526..7317,7318..7386))
                     /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     7149..7168
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7387..7491)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg agctccaccg
      661 cggtggcggc cgctctagaa ttccattgca tacgttgtat ccatatcata atatgtacat
      721 ttatattggc tcatgtccaa cattaccgcc atgttgacat tgattattga ctagttatta
      781 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      841 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      901 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      961 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     1021 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     1081 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     1141 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     1201 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     1261 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     1321 ggtctatata agcagagctc gtttagtgaa ccgcagtcgc tctgcggaga ggctggcaga
     1381 ttgagccctg ggaggttctc tccagcacta gcaggtagag cctgggtgtt ccctgctaga
     1441 ctctcaccag cacttggcca gtgctgggca gagtggctcc acgcttgctt gcttaaagac
     1501 ctcttcaata aagctgccat tttagaagta agccagtgtg tgttcccatc tctcctagtc
     1561 gccgcctggt caactcggta ctcggtaata agaagaccct ggtctgttag gaccctttct
     1621 gctttgagaa accgaagcag gaaaatccct agcagattgg cgcccgaaca gggacttgaa
     1681 ggagagtgag agactcctga gtacggctga gtgaaggcag taagggcggc aggaaccaac
     1741 cacgacggag tgctcctata aaggcgcggg tcggtaccag acggcgtgag gagcgggaga
     1801 ggaggaggcc tccggttgca ggtaagtgca acacaaaaaa gaaatagctg tcttgttatc
     1861 caggaaggga taataagata gagtgggaga tgggcgcgag aaactccgtc ttgtcaggga
     1921 agaaagcaga tgaattgaaa aaattattaa tcgcatgaat tttaaaagaa ggggaggaat
     1981 aggggatatg actccagcag aaagattaat taacatgatc actacagaac aagaaataca
     2041 atttcaacaa tcaaaaaact caaaatttaa aaattttcgg gtctattaca gagctcacgc
     2101 gtgattggag ttgggagatt ataaattagt agagatcact ccgattggct tggcccccac
     2161 agatgtgaag aggtacacta ctggtggcac ctcaagaaat aaaagagggg tctttgtgct
     2221 agggttcttg ggttttctcg caacggcagg ttctgcaatg ggcgcggcgt cgttgacgct
     2281 gaccgctcag tcccggactt tattggctgg gatagtgcag caacagcaac agctgttgga
     2341 cgtggtcaag agacaacaag aattgttgcg actgaccgtc tggggaacaa agaacctcca
     2401 gactagggtc actgccatcg agaagtactt aaaggaccag gcgcagctaa atgcttgggg
     2461 atgtgcgttt agacaagtct gccacactac tgtaccatgg ccaaatgcaa gtctaacacc
     2521 agactggaac aatgatactt ggcaagagtg ggagcgaaag gttgacttct tggaggaaaa
     2581 tataacagcc ctcctagaag aggcacaaat tcaacaagag aagaacatgt atgaattaca
     2641 aaagttgaat agctgggatg tgtttggcaa ttggtttgac cttgcttctt ggataaagta
     2701 tatacaatat ggaatttatg tagttgtagg agtaatactg ttaagaatag tgatctatat
     2761 agtacaaatg ctagctaagt taaggcaggg gtataggcca gtgttctctt ccccaccctc
     2821 ttatttccag tagactcata cccaacagga cccggcactg ccaaccagag aaggcaaaga
     2881 aggacggtgg agaaggcggt ggatctgcta gcactagtta ttaatcgtac gattaactgc
     2941 agccccgata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc
     3001 cacctgtagg tttggcaagc tagctgcagt aacgccattt tgcaaggcat ggaaaaatac
     3061 caaaccaaga atagagaagt tcagatcaag ggcgggtaca tgaaaatagc taacgttggg
     3121 ccaaacagga tatctgcggt gagcagtttc ggccccggcc cggggccaag aacagatggt
     3181 caccgcagtt tcggccccgg cccgaggcca agaacagatg gtccccagat atggcccaac
     3241 cctcagcagt ttcttaagac ccatcagatg tttccaggct cccccaagga cctgaaatga
     3301 ccctgcgcct tatttgaatt aaccaatcag cctgcttctc gcttctgttc gcgcgcttct
     3361 gcttcccgag ctctataaaa gagctcacaa cccctcactc ggcgcgccag tcctccgaca
     3421 gactgagtcg cccgggggac cgggagcaga tctaccggta tggccaagcc tttgtctcaa
     3481 gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa
     3541 gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat
     3601 gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct
     3661 gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg
     3721 agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata
     3781 gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt
     3841 tatgtgtggg agggctaact cgagatccaa gaaggtatat tgctgttgac agtgagcggc
     3901 ctaagacagt gcctatacat tagtgaagcc acagatgtaa tgtataggca ctgtcttagg
     3961 gtgcctactg cctcggactt caagggctac gatgtcgact tggatccatc gatggatcta
     4021 attcccgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     4081 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     4141 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     4201 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     4261 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     4321 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     4381 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     4441 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     4501 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     4561 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     4621 cgtcgagttt tataaaagaa aaggggggac tggaagggat ttattacagt gcaagaagac
     4681 atagaatctt agacatgtac ttagaaaagg aagaaggcat cataccagat tggcaggatt
     4741 acacctcagg accaggaatt agatacccaa agacatttgg ctggctatgg aaattagtcc
     4801 ctgtaaatgt atcagatgag gcacaggagg atgagaggca ttatttaatg cagccagctc
     4861 aaacttccaa gtgggatgac ccttggggag aggttctagc gtggaagttt gatccaactc
     4921 tagcctacac ttatgaggca tatgctagat acccagaaga gttggaagca agtcaggcct
     4981 gtcagaactg catttcgctc tgtattcagt cgctctgcgg agaggctggc agattgagcc
     5041 ctgggaggtt ctctccagca ctagcaggta gagcctgggt gttccctgct agactctcac
     5101 cagcacttgg ccagtgctgg gcagagtggc tccacgcttg cttgcttaaa gacctcttca
     5161 ataaagctgc cattttagaa gtaagccagt gtgtgttccc atctctccta gtcgccgcct
     5221 ggtcaactcg gtactcggta ataagaagac cctggtctgt taggaccctt tctgctttga
     5281 gaaaccgaag caggaaaatc cctagcatgg taccagcttt tgttcccttt agtgagggtt
     5341 aatttcgagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5401 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5461 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5521 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5581 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5641 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5701 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5761 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5821 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5881 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5941 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6001 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6061 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6121 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6181 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6241 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6301 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6361 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6421 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6481 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6541 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6601 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6661 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6721 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6781 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6841 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6901 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6961 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7021 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7081 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7141 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7201 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7261 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7321 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7381 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7441 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7501 ccgaaaagtg ccac
//
