LOCUS       sequence-337047-        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: miR5 MmATP10B
FEATURES             Location/Qualifiers
     source          1..7514
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(118..139)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     330..349
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     563..585
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     577..594
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     578..594
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     601..620
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        601..619
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        734..1113
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1114..1317
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1268..1288
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3007..3414
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             3438..3836
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     ncRNA           3901..3915
                     /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    4008..4596
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4061..4081)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4479..4490)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(5304..5322)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5343..5359)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5343..5359)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5356..5378)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5367..5383
                     /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(5391..5397,5398..5415,5416..5421))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5436..5457
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5574..5591)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5745..6333)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5825..5844)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6504..7295,7296..7364))
                     /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     7127..7146
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7365..7469)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(7495..7514,1..436))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
       61 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
      121 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
      181 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
      241 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
      301 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
      361 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
      421 ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg
      481 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
      541 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg
      601 taatacgact cactataggg cgaattggag ctccaccgcg gtggcggccg ctctagaatt
      661 ccattgcata cgttgtatcc atatcataat atgtacattt atattggctc atgtccaaca
      721 ttaccgccat gttgacattg attattgact agttattaat agtaatcaat tacggggtca
      781 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      841 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      901 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      961 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     1021 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     1081 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     1141 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     1201 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     1261 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
     1321 ttagtgaacc gcagtcgctc tgcggagagg ctggcagatt gagccctggg aggttctctc
     1381 cagcactagc aggtagagcc tgggtgttcc ctgctagact ctcaccagca cttggccagt
     1441 gctgggcaga gtggctccac gcttgcttgc ttaaagacct cttcaataaa gctgccattt
     1501 tagaagtaag ccagtgtgtg ttcccatctc tcctagtcgc cgcctggtca actcggtact
     1561 cggtaataag aagaccctgg tctgttagga ccctttctgc tttgagaaac cgaagcagga
     1621 aaatccctag cagattggcg cccgaacagg gacttgaagg agagtgagag actcctgagt
     1681 acggctgagt gaaggcagta agggcggcag gaaccaacca cgacggagtg ctcctataaa
     1741 ggcgcgggtc ggtaccagac ggcgtgagga gcgggagagg aggaggcctc cggttgcagg
     1801 taagtgcaac acaaaaaaga aatagctgtc ttgttatcca ggaagggata ataagataga
     1861 gtgggagatg ggcgcgagaa actccgtctt gtcagggaag aaagcagatg aattgaaaaa
     1921 attattaatc gcatgaattt taaaagaagg ggaggaatag gggatatgac tccagcagaa
     1981 agattaatta acatgatcac tacagaacaa gaaatacaat ttcaacaatc aaaaaactca
     2041 aaatttaaaa attttcgggt ctattacaga gctcacgcgt gattggagtt gggagattat
     2101 aaattagtag agatcactcc gattggcttg gcccccacag atgtgaagag gtacactact
     2161 ggtggcacct caagaaataa aagaggggtc tttgtgctag ggttcttggg ttttctcgca
     2221 acggcaggtt ctgcaatggg cgcggcgtcg ttgacgctga ccgctcagtc ccggacttta
     2281 ttggctggga tagtgcagca acagcaacag ctgttggacg tggtcaagag acaacaagaa
     2341 ttgttgcgac tgaccgtctg gggaacaaag aacctccaga ctagggtcac tgccatcgag
     2401 aagtacttaa aggaccaggc gcagctaaat gcttggggat gtgcgtttag acaagtctgc
     2461 cacactactg taccatggcc aaatgcaagt ctaacaccag actggaacaa tgatacttgg
     2521 caagagtggg agcgaaaggt tgacttcttg gaggaaaata taacagccct cctagaagag
     2581 gcacaaattc aacaagagaa gaacatgtat gaattacaaa agttgaatag ctgggatgtg
     2641 tttggcaatt ggtttgacct tgcttcttgg ataaagtata tacaatatgg aatttatgta
     2701 gttgtaggag taatactgtt aagaatagtg atctatatag tacaaatgct agctaagtta
     2761 aggcaggggt ataggccagt gttctcttcc ccaccctctt atttccagta gactcatacc
     2821 caacaggacc cggcactgcc aaccagagaa ggcaaagaag gacggtggag aaggcggtgg
     2881 atctgctagc actagttatt aatcgtacga ttaactgcag ccccgataaa ataaaagatt
     2941 ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta
     3001 gctgcagtaa cgccattttg caaggcatgg aaaaatacca aaccaagaat agagaagttc
     3061 agatcaaggg cgggtacatg aaaatagcta acgttgggcc aaacaggata tctgcggtga
     3121 gcagtttcgg ccccggcccg gggccaagaa cagatggtca ccgcagtttc ggccccggcc
     3181 cgaggccaag aacagatggt ccccagatat ggcccaaccc tcagcagttt cttaagaccc
     3241 atcagatgtt tccaggctcc cccaaggacc tgaaatgacc ctgcgcctta tttgaattaa
     3301 ccaatcagcc tgcttctcgc ttctgttcgc gcgcttctgc ttcccgagct ctataaaaga
     3361 gctcacaacc cctcactcgg cgcgccagtc ctccgacaga ctgagtcgcc cgggggaccg
     3421 ggagcagatc taccggtatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa
     3481 gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag
     3541 ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac
     3601 cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt
     3661 gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac
     3721 aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc
     3781 cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaactcg
     3841 agatccaaga aggtatattg ctgttgacag tgagcgacct ggtgattctg aactggatta
     3901 gtgaagccac agatgtaatc cagttcagaa tcaccaggat gcctactgcc tcggacttca
     3961 agggctacga tgtcgacttg gatccatcga tggatctaat tcccgataat caacctctgg
     4021 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     4081 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     4141 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     4201 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     4261 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     4321 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     4381 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     4441 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     4501 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     4561 agacgagtcg gatctccctt tgggccgcct ccccgcatcg tcgagtttta taaaagaaaa
     4621 ggggggactg gaagggattt attacagtgc aagaagacat agaatcttag acatgtactt
     4681 agaaaaggaa gaaggcatca taccagattg gcaggattac acctcaggac caggaattag
     4741 atacccaaag acatttggct ggctatggaa attagtccct gtaaatgtat cagatgaggc
     4801 acaggaggat gagaggcatt atttaatgca gccagctcaa acttccaagt gggatgaccc
     4861 ttggggagag gttctagcgt ggaagtttga tccaactcta gcctacactt atgaggcata
     4921 tgctagatac ccagaagagt tggaagcaag tcaggcctgt cagaactgca tttcgctctg
     4981 tattcagtcg ctctgcggag aggctggcag attgagccct gggaggttct ctccagcact
     5041 agcaggtaga gcctgggtgt tccctgctag actctcacca gcacttggcc agtgctgggc
     5101 agagtggctc cacgcttgct tgcttaaaga cctcttcaat aaagctgcca ttttagaagt
     5161 aagccagtgt gtgttcccat ctctcctagt cgccgcctgg tcaactcggt actcggtaat
     5221 aagaagaccc tggtctgtta ggaccctttc tgctttgaga aaccgaagca ggaaaatccc
     5281 tagcatggta ccagcttttg ttccctttag tgagggttaa tttcgagctt ggcgtaatca
     5341 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5401 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5461 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5521 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5581 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5641 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5701 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5761 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5821 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5881 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5941 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6001 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6061 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6121 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6181 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6241 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6301 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6361 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6421 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6481 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6541 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6601 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6661 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6721 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6781 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6841 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6901 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6961 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7021 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7081 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7141 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7201 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7261 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7321 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7381 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7441 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
     7501 taagcgttaa tatt
//
