LOCUS       sequence-337044-        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: miR3 Mm ATP13A2
FEATURES             Location/Qualifiers
     source          1..7514
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(37..492)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(174..195)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     386..405
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     619..641
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     633..650
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     634..650
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     657..676
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        657..675
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        790..1169
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1170..1373
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1324..1344
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3063..3470
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             3494..3892
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     ncRNA           3957..3971
                     /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    4064..4652
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4117..4137)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4535..4546)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(5360..5378)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5399..5415)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5399..5415)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5412..5434)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5423..5439
                     /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(5447..5453,5454..5471,5472..5477))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5492..5513
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5630..5647)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5801..6389)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5881..5900)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6560..7351,7352..7420))
                     /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     7183..7202
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7421..7514,1..11))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
       61 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
      121 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
      181 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
      241 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
      301 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
      361 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
      421 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
      481 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
      541 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
      601 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattgtaat
      661 acgactcact atagggcgaa ttggagctcc accgcggtgg cggccgctct agaattccat
      721 tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt ccaacattac
      781 cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      841 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      901 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      961 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     1021 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     1081 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1141 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     1201 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     1261 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     1321 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
     1381 tgaaccgcag tcgctctgcg gagaggctgg cagattgagc cctgggaggt tctctccagc
     1441 actagcaggt agagcctggg tgttccctgc tagactctca ccagcacttg gccagtgctg
     1501 ggcagagtgg ctccacgctt gcttgcttaa agacctcttc aataaagctg ccattttaga
     1561 agtaagccag tgtgtgttcc catctctcct agtcgccgcc tggtcaactc ggtactcggt
     1621 aataagaaga ccctggtctg ttaggaccct ttctgctttg agaaaccgaa gcaggaaaat
     1681 ccctagcaga ttggcgcccg aacagggact tgaaggagag tgagagactc ctgagtacgg
     1741 ctgagtgaag gcagtaaggg cggcaggaac caaccacgac ggagtgctcc tataaaggcg
     1801 cgggtcggta ccagacggcg tgaggagcgg gagaggagga ggcctccggt tgcaggtaag
     1861 tgcaacacaa aaaagaaata gctgtcttgt tatccaggaa gggataataa gatagagtgg
     1921 gagatgggcg cgagaaactc cgtcttgtca gggaagaaag cagatgaatt gaaaaaatta
     1981 ttaatcgcat gaattttaaa agaaggggag gaatagggga tatgactcca gcagaaagat
     2041 taattaacat gatcactaca gaacaagaaa tacaatttca acaatcaaaa aactcaaaat
     2101 ttaaaaattt tcgggtctat tacagagctc acgcgtgatt ggagttggga gattataaat
     2161 tagtagagat cactccgatt ggcttggccc ccacagatgt gaagaggtac actactggtg
     2221 gcacctcaag aaataaaaga ggggtctttg tgctagggtt cttgggtttt ctcgcaacgg
     2281 caggttctgc aatgggcgcg gcgtcgttga cgctgaccgc tcagtcccgg actttattgg
     2341 ctgggatagt gcagcaacag caacagctgt tggacgtggt caagagacaa caagaattgt
     2401 tgcgactgac cgtctgggga acaaagaacc tccagactag ggtcactgcc atcgagaagt
     2461 acttaaagga ccaggcgcag ctaaatgctt ggggatgtgc gtttagacaa gtctgccaca
     2521 ctactgtacc atggccaaat gcaagtctaa caccagactg gaacaatgat acttggcaag
     2581 agtgggagcg aaaggttgac ttcttggagg aaaatataac agccctccta gaagaggcac
     2641 aaattcaaca agagaagaac atgtatgaat tacaaaagtt gaatagctgg gatgtgtttg
     2701 gcaattggtt tgaccttgct tcttggataa agtatataca atatggaatt tatgtagttg
     2761 taggagtaat actgttaaga atagtgatct atatagtaca aatgctagct aagttaaggc
     2821 aggggtatag gccagtgttc tcttccccac cctcttattt ccagtagact catacccaac
     2881 aggacccggc actgccaacc agagaaggca aagaaggacg gtggagaagg cggtggatct
     2941 gctagcacta gttattaatc gtacgattaa ctgcagcccc gataaaataa aagattttat
     3001 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctg
     3061 cagtaacgcc attttgcaag gcatggaaaa ataccaaacc aagaatagag aagttcagat
     3121 caagggcggg tacatgaaaa tagctaacgt tgggccaaac aggatatctg cggtgagcag
     3181 tttcggcccc ggcccggggc caagaacaga tggtcaccgc agtttcggcc ccggcccgag
     3241 gccaagaaca gatggtcccc agatatggcc caaccctcag cagtttctta agacccatca
     3301 gatgtttcca ggctccccca aggacctgaa atgaccctgc gccttatttg aattaaccaa
     3361 tcagcctgct tctcgcttct gttcgcgcgc ttctgcttcc cgagctctat aaaagagctc
     3421 acaacccctc actcggcgcg ccagtcctcc gacagactga gtcgcccggg ggaccgggag
     3481 cagatctacc ggtatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc
     3541 aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct
     3601 ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg
     3661 tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat
     3721 cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt
     3781 gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac
     3841 ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct aactcgagat
     3901 ccaagaaggt atattgctgt tgacagtgag cggcacgccg aaacactcgt tatatagtga
     3961 agccacagat gtatataacg agtgtttcgg cgtggtgcct actgcctcgg acttcaaggg
     4021 ctacgatgtc gacttggatc catcgatgga tctaattccc gataatcaac ctctggatta
     4081 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     4141 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     4201 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     4261 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     4321 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     4381 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     4441 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     4501 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     4561 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     4621 gagtcggatc tccctttggg ccgcctcccc gcatcgtcga gttttataaa agaaaagggg
     4681 ggactggaag ggatttatta cagtgcaaga agacatagaa tcttagacat gtacttagaa
     4741 aaggaagaag gcatcatacc agattggcag gattacacct caggaccagg aattagatac
     4801 ccaaagacat ttggctggct atggaaatta gtccctgtaa atgtatcaga tgaggcacag
     4861 gaggatgaga ggcattattt aatgcagcca gctcaaactt ccaagtggga tgacccttgg
     4921 ggagaggttc tagcgtggaa gtttgatcca actctagcct acacttatga ggcatatgct
     4981 agatacccag aagagttgga agcaagtcag gcctgtcaga actgcatttc gctctgtatt
     5041 cagtcgctct gcggagaggc tggcagattg agccctggga ggttctctcc agcactagca
     5101 ggtagagcct gggtgttccc tgctagactc tcaccagcac ttggccagtg ctgggcagag
     5161 tggctccacg cttgcttgct taaagacctc ttcaataaag ctgccatttt agaagtaagc
     5221 cagtgtgtgt tcccatctct cctagtcgcc gcctggtcaa ctcggtactc ggtaataaga
     5281 agaccctggt ctgttaggac cctttctgct ttgagaaacc gaagcaggaa aatccctagc
     5341 atggtaccag cttttgttcc ctttagtgag ggttaatttc gagcttggcg taatcatggt
     5401 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5461 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5521 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5581 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5641 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5701 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5761 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5821 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5881 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5941 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6001 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6061 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6121 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6181 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6241 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6301 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6361 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6421 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6481 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6541 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6601 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6661 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6721 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6781 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6841 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6901 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6961 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7021 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7081 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7141 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7201 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7261 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7321 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7381 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7441 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7501 aaaataaaca aata
//
