LOCUS       sequence-337049-        7514 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transfer plasmid for lentiviral vector production with miR for Mm 
            ATP13A2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7514)
  AUTHORS   van Veen S, Martin S, Van den Haute C, Benoy V, Lyons J, Vanhoutte 
            R, Kahler JP, Decuypere JP, Gelders G, Lambie E, Zielich J, Swinnen 
            JV, Annaert W, Agostinis P, Ghesquiere B, Verhelst S, Baekelandt V, 
            Eggermont J, Vangheluwe P
  TITLE     ATP13A2 deficiency disrupts lysosomal polyamine export.
  JOURNAL   Nature. 2020 Feb;578(7795):419-424. doi: 10.1038/s41586-020-1968-7. 
            Epub 2020 Jan 29.
  PUBMED    31996848
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: 
            "10.1038/s41586-020-1968-7"; journalName: "Nature"; date: "2020-02";
            volume: "578"; issue: "7795"; pages: "419-424"
COMMENT     Sequence Label: miR5 Mm ATP13A2
FEATURES             Location/Qualifiers
     source          1..7514
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        73..452
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        453..656
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     607..627
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2346..2753
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             2777..3175
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     ncRNA           3240..3254
                     /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    3347..3935
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3400..3420)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3818..3829)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(4643..4661)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4682..4698)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4682..4698)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4695..4717)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4706..4722
                     /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(4730..4736,4737..4754,4755..4760))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4775..4796
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4913..4930)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5084..5672)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5164..5183)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5843..6634,6635..6703))
                     /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     6466..6485
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6704..6808)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6834..7289)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6971..6992)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7183..7202
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7416..7438
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7430..7447
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7431..7447
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7454..7473
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7454..7472
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cattgcatac gttgtatcca tatcataata tgtacattta tattggctca tgtccaacat
       61 taccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
      121 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      181 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      241 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      301 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      361 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      421 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
      481 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      541 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      601 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt
      661 tagtgaaccg cagtcgctct gcggagaggc tggcagattg agccctggga ggttctctcc
      721 agcactagca ggtagagcct gggtgttccc tgctagactc tcaccagcac ttggccagtg
      781 ctgggcagag tggctccacg cttgcttgct taaagacctc ttcaataaag ctgccatttt
      841 agaagtaagc cagtgtgtgt tcccatctct cctagtcgcc gcctggtcaa ctcggtactc
      901 ggtaataaga agaccctggt ctgttaggac cctttctgct ttgagaaacc gaagcaggaa
      961 aatccctagc agattggcgc ccgaacaggg acttgaagga gagtgagaga ctcctgagta
     1021 cggctgagtg aaggcagtaa gggcggcagg aaccaaccac gacggagtgc tcctataaag
     1081 gcgcgggtcg gtaccagacg gcgtgaggag cgggagagga ggaggcctcc ggttgcaggt
     1141 aagtgcaaca caaaaaagaa atagctgtct tgttatccag gaagggataa taagatagag
     1201 tgggagatgg gcgcgagaaa ctccgtcttg tcagggaaga aagcagatga attgaaaaaa
     1261 ttattaatcg catgaatttt aaaagaaggg gaggaatagg ggatatgact ccagcagaaa
     1321 gattaattaa catgatcact acagaacaag aaatacaatt tcaacaatca aaaaactcaa
     1381 aatttaaaaa ttttcgggtc tattacagag ctcacgcgtg attggagttg ggagattata
     1441 aattagtaga gatcactccg attggcttgg cccccacaga tgtgaagagg tacactactg
     1501 gtggcacctc aagaaataaa agaggggtct ttgtgctagg gttcttgggt tttctcgcaa
     1561 cggcaggttc tgcaatgggc gcggcgtcgt tgacgctgac cgctcagtcc cggactttat
     1621 tggctgggat agtgcagcaa cagcaacagc tgttggacgt ggtcaagaga caacaagaat
     1681 tgttgcgact gaccgtctgg ggaacaaaga acctccagac tagggtcact gccatcgaga
     1741 agtacttaaa ggaccaggcg cagctaaatg cttggggatg tgcgtttaga caagtctgcc
     1801 acactactgt accatggcca aatgcaagtc taacaccaga ctggaacaat gatacttggc
     1861 aagagtggga gcgaaaggtt gacttcttgg aggaaaatat aacagccctc ctagaagagg
     1921 cacaaattca acaagagaag aacatgtatg aattacaaaa gttgaatagc tgggatgtgt
     1981 ttggcaattg gtttgacctt gcttcttgga taaagtatat acaatatgga atttatgtag
     2041 ttgtaggagt aatactgtta agaatagtga tctatatagt acaaatgcta gctaagttaa
     2101 ggcaggggta taggccagtg ttctcttccc caccctctta tttccagtag actcataccc
     2161 aacaggaccc ggcactgcca accagagaag gcaaagaagg acggtggaga aggcggtgga
     2221 tctgctagca ctagttatta atcgtacgat taactgcagc cccgataaaa taaaagattt
     2281 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     2341 ctgcagtaac gccattttgc aaggcatgga aaaataccaa accaagaata gagaagttca
     2401 gatcaagggc gggtacatga aaatagctaa cgttgggcca aacaggatat ctgcggtgag
     2461 cagtttcggc cccggcccgg ggccaagaac agatggtcac cgcagtttcg gccccggccc
     2521 gaggccaaga acagatggtc cccagatatg gcccaaccct cagcagtttc ttaagaccca
     2581 tcagatgttt ccaggctccc ccaaggacct gaaatgaccc tgcgccttat ttgaattaac
     2641 caatcagcct gcttctcgct tctgttcgcg cgcttctgct tcccgagctc tataaaagag
     2701 ctcacaaccc ctcactcggc gcgccagtcc tccgacagac tgagtcgccc gggggaccgg
     2761 gagcagatct accggtatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag
     2821 agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc
     2881 tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc
     2941 ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg
     3001 tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca
     3061 ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc
     3121 gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctaactcga
     3181 gatccaagaa ggtatattgc tgttgacagt gagcgggccg aaacactcgt tatagaatag
     3241 tgaagccaca gatgtattct ataacgagtg tttcggcgtg cctactgcct cggacttcaa
     3301 gggctacgat gtcgacttgg atccatcgat ggatctaatt cccgataatc aacctctgga
     3361 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3421 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3481 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3541 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3601 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3661 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3721 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     3781 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3841 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3901 gacgagtcgg atctcccttt gggccgcctc cccgcatcgt cgagttttat aaaagaaaag
     3961 gggggactgg aagggattta ttacagtgca agaagacata gaatcttaga catgtactta
     4021 gaaaaggaag aaggcatcat accagattgg caggattaca cctcaggacc aggaattaga
     4081 tacccaaaga catttggctg gctatggaaa ttagtccctg taaatgtatc agatgaggca
     4141 caggaggatg agaggcatta tttaatgcag ccagctcaaa cttccaagtg ggatgaccct
     4201 tggggagagg ttctagcgtg gaagtttgat ccaactctag cctacactta tgaggcatat
     4261 gctagatacc cagaagagtt ggaagcaagt caggcctgtc agaactgcat ttcgctctgt
     4321 attcagtcgc tctgcggaga ggctggcaga ttgagccctg ggaggttctc tccagcacta
     4381 gcaggtagag cctgggtgtt ccctgctaga ctctcaccag cacttggcca gtgctgggca
     4441 gagtggctcc acgcttgctt gcttaaagac ctcttcaata aagctgccat tttagaagta
     4501 agccagtgtg tgttcccatc tctcctagtc gccgcctggt caactcggta ctcggtaata
     4561 agaagaccct ggtctgttag gaccctttct gctttgagaa accgaagcag gaaaatccct
     4621 agcatggtac cagcttttgt tccctttagt gagggttaat ttcgagcttg gcgtaatcat
     4681 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4741 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4801 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4861 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4921 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4981 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5041 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5101 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5161 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5221 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5281 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5341 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5401 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5461 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5521 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5581 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5641 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5701 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5761 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5821 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5881 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5941 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6001 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6061 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6121 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6181 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6241 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6301 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6361 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6421 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6481 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6541 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6601 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6661 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6721 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6781 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
     6841 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     6901 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     6961 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     7021 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     7081 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     7141 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     7201 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     7261 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
     7321 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     7381 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     7441 ggccagtgaa ttgtaatacg actcactata gggcgaattg gagctccacc gcggtggcgg
     7501 ccgctctaga attc
//
