LOCUS       sequence-337046-        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: miR2 HsATP10B
FEATURES             Location/Qualifiers
     source          1..7514
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(47..502)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(184..205)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     396..415
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     629..651
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     643..660
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     644..660
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     667..686
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        667..685
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        800..1179
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1180..1383
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1334..1354
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3073..3480
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             3504..3902
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     ncRNA           3967..3981
                     /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    4074..4662
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4127..4147)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4545..4556)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(5370..5388)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5409..5425)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5409..5425)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5422..5444)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5433..5449
                     /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(5457..5463,5464..5481,5482..5487))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5502..5523
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5640..5657)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5811..6399)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5891..5910)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6570..7361,7362..7430))
                     /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     7193..7212
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7431..7514,1..21))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
       61 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
      121 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
      181 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
      241 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
      301 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
      361 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
      421 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
      481 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
      541 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
      601 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
      661 gaattgtaat acgactcact atagggcgaa ttggagctcc accgcggtgg cggccgctct
      721 agaattccat tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt
      781 ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg
      841 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      901 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      961 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     1021 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     1081 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     1141 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     1201 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     1261 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     1321 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     1381 gctcgtttag tgaaccgcag tcgctctgcg gagaggctgg cagattgagc cctgggaggt
     1441 tctctccagc actagcaggt agagcctggg tgttccctgc tagactctca ccagcacttg
     1501 gccagtgctg ggcagagtgg ctccacgctt gcttgcttaa agacctcttc aataaagctg
     1561 ccattttaga agtaagccag tgtgtgttcc catctctcct agtcgccgcc tggtcaactc
     1621 ggtactcggt aataagaaga ccctggtctg ttaggaccct ttctgctttg agaaaccgaa
     1681 gcaggaaaat ccctagcaga ttggcgcccg aacagggact tgaaggagag tgagagactc
     1741 ctgagtacgg ctgagtgaag gcagtaaggg cggcaggaac caaccacgac ggagtgctcc
     1801 tataaaggcg cgggtcggta ccagacggcg tgaggagcgg gagaggagga ggcctccggt
     1861 tgcaggtaag tgcaacacaa aaaagaaata gctgtcttgt tatccaggaa gggataataa
     1921 gatagagtgg gagatgggcg cgagaaactc cgtcttgtca gggaagaaag cagatgaatt
     1981 gaaaaaatta ttaatcgcat gaattttaaa agaaggggag gaatagggga tatgactcca
     2041 gcagaaagat taattaacat gatcactaca gaacaagaaa tacaatttca acaatcaaaa
     2101 aactcaaaat ttaaaaattt tcgggtctat tacagagctc acgcgtgatt ggagttggga
     2161 gattataaat tagtagagat cactccgatt ggcttggccc ccacagatgt gaagaggtac
     2221 actactggtg gcacctcaag aaataaaaga ggggtctttg tgctagggtt cttgggtttt
     2281 ctcgcaacgg caggttctgc aatgggcgcg gcgtcgttga cgctgaccgc tcagtcccgg
     2341 actttattgg ctgggatagt gcagcaacag caacagctgt tggacgtggt caagagacaa
     2401 caagaattgt tgcgactgac cgtctgggga acaaagaacc tccagactag ggtcactgcc
     2461 atcgagaagt acttaaagga ccaggcgcag ctaaatgctt ggggatgtgc gtttagacaa
     2521 gtctgccaca ctactgtacc atggccaaat gcaagtctaa caccagactg gaacaatgat
     2581 acttggcaag agtgggagcg aaaggttgac ttcttggagg aaaatataac agccctccta
     2641 gaagaggcac aaattcaaca agagaagaac atgtatgaat tacaaaagtt gaatagctgg
     2701 gatgtgtttg gcaattggtt tgaccttgct tcttggataa agtatataca atatggaatt
     2761 tatgtagttg taggagtaat actgttaaga atagtgatct atatagtaca aatgctagct
     2821 aagttaaggc aggggtatag gccagtgttc tcttccccac cctcttattt ccagtagact
     2881 catacccaac aggacccggc actgccaacc agagaaggca aagaaggacg gtggagaagg
     2941 cggtggatct gctagcacta gttattaatc gtacgattaa ctgcagcccc gataaaataa
     3001 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     3061 aagctagctg cagtaacgcc attttgcaag gcatggaaaa ataccaaacc aagaatagag
     3121 aagttcagat caagggcggg tacatgaaaa tagctaacgt tgggccaaac aggatatctg
     3181 cggtgagcag tttcggcccc ggcccggggc caagaacaga tggtcaccgc agtttcggcc
     3241 ccggcccgag gccaagaaca gatggtcccc agatatggcc caaccctcag cagtttctta
     3301 agacccatca gatgtttcca ggctccccca aggacctgaa atgaccctgc gccttatttg
     3361 aattaaccaa tcagcctgct tctcgcttct gttcgcgcgc ttctgcttcc cgagctctat
     3421 aaaagagctc acaacccctc actcggcgcg ccagtcctcc gacagactga gtcgcccggg
     3481 ggaccgggag cagatctacc ggtatggcca agcctttgtc tcaagaagaa tccaccctca
     3541 ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca
     3601 gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg
     3661 ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc
     3721 tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt
     3781 gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg
     3841 gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct
     3901 aactcgagat ccaagaaggt atattgctgt tgacagtgag cgttgattca agctgctgat
     3961 attgtagtga agccacagat gtacaatatc agcagcttga atcattgcct actgcctcgg
     4021 acttcaaggg ctacgatgtc gacttggatc catcgatgga tctaattccc gataatcaac
     4081 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     4141 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     4201 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     4261 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     4321 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     4381 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     4441 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     4501 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     4561 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     4621 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgtcga gttttataaa
     4681 agaaaagggg ggactggaag ggatttatta cagtgcaaga agacatagaa tcttagacat
     4741 gtacttagaa aaggaagaag gcatcatacc agattggcag gattacacct caggaccagg
     4801 aattagatac ccaaagacat ttggctggct atggaaatta gtccctgtaa atgtatcaga
     4861 tgaggcacag gaggatgaga ggcattattt aatgcagcca gctcaaactt ccaagtggga
     4921 tgacccttgg ggagaggttc tagcgtggaa gtttgatcca actctagcct acacttatga
     4981 ggcatatgct agatacccag aagagttgga agcaagtcag gcctgtcaga actgcatttc
     5041 gctctgtatt cagtcgctct gcggagaggc tggcagattg agccctggga ggttctctcc
     5101 agcactagca ggtagagcct gggtgttccc tgctagactc tcaccagcac ttggccagtg
     5161 ctgggcagag tggctccacg cttgcttgct taaagacctc ttcaataaag ctgccatttt
     5221 agaagtaagc cagtgtgtgt tcccatctct cctagtcgcc gcctggtcaa ctcggtactc
     5281 ggtaataaga agaccctggt ctgttaggac cctttctgct ttgagaaacc gaagcaggaa
     5341 aatccctagc atggtaccag cttttgttcc ctttagtgag ggttaatttc gagcttggcg
     5401 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5461 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5521 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5581 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5641 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5701 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5761 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5821 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5881 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5941 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6001 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6061 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6121 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6181 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6241 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6301 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6361 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6421 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6481 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6541 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6601 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6661 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6721 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6781 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6841 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6901 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6961 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7021 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7081 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7141 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7201 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7261 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7321 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7381 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7441 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7501 tgtatttaga aaaa
//
