LOCUS       sequence-181059-        4534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of M1 N184X spastin in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4534)
  AUTHORS   Solowska JM, Rao AN, Baas PW
  TITLE     Studies on truncating mutations of SPAST associated with Hereditary 
            Spastic Paraplegia indicate greater accumulation and toxicity of the
            M1 isoform of spastin.
  JOURNAL   Mol Biol Cell. 2017 May 11. pii: mbc.E17-01-0047. doi: 
            10.1091/mbc.E17-01-0047.
  PUBMED    28495799
REFERENCE   2  (bases 1 to 4534)
  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"; journalName: "Mol Biol 
            Cell. 2017 May 11. pii: mbc.E17-01-0047. doi: 
            10.1091/mbc.E17-01-0047."
COMMENT     Sequence Label: pTRE/SP M1 N184X
FEATURES             Location/Qualifiers
     source          1..4534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..315
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    9..27
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    45..63
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    80..98
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    116..134
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    152..170
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    187..205
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    223..241
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     288..312
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     polyA_signal    2445..2526
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2482..2501)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2537..2554)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2708..3296)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2788..2807)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3467..4258,4259..4327))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4090..4109
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4328..4432)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4500..4518
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gagtttactc cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag
       61 agaacgatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag tttactccct
      121 atcagtgata gagaacgtat gtcgagttta ctccctatca gtgatagaga acgtatgtcg
      181 agtttatccc tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag
      241 aacgtatgtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg
      301 aaccgtcaga tcgcctggag aattcgagct cggtacccat cgcggcggcg gcagtgagag
      361 gccaccatga attctccggg tggacgaggg aagaagaaag gctccggcgg cgccagcaac
      421 ccggtgcctc ccaggcctcc gcccccttgc ctggcccccg cccctcccgc cgccgggccg
      481 gcccctccgc ccgagtcgcc gcataagcgg aacctgtact atttctccta cccgctgttt
      541 gtaggcttcg cgctgctgcg tttggtcgcc ttccacctgg ggctcctctt cgtgtggctc
      601 tgccagcgct tctcccgcgc cctcatggca gccaagagga gctccggggc cgcgccagca
      661 cctgcctcgg cctcggcccc ggcgccggtg ccgggcggcg aggccgagcg cgtccgagtc
      721 ttccacaaac aggccttcga gtacatctcc attgccctgc gcatcgatga ggatgagaaa
      781 gcaggacaga aggagcaagc tgtggaatgg tataagaaag gtattgaaga actggaaaaa
      841 ggaatagctg ttatagttac aggacaaggt gaacagtgtg aaagagctag acgccttcaa
      901 gctaaaatga tgacttaatt tggttatggc caaggaccgc ttacaacttc tagagaagat
      961 gcaaccagtt ttgccatttt ccaagtcaca aacggacgtc tataatgaca gtactaactt
     1021 ggcatgccgc aatggacatc tccagtcaga aagtggagct gttccaaaaa gaaaagaccc
     1081 cttaacacac actagtaatt cactgcctcg ttcaaaaaca gttatgaaaa ctggatctgc
     1141 aggcctttca ggccaccata gagcacctag ttacagtggt ttatccatgg tttctggagt
     1201 gaaacaggga tctggtcctg ctcctaccac tcataagggt actccgaaaa caaataggac
     1261 aaataaacct tctaccccta caactgctac tcgtaagaaa aaagacttga agaattttag
     1321 gaatgtggac agcaaccttg ctaaccttat aatgaatgaa attgtggaca atggaacagc
     1381 tgttaaattt gatgatatag ctggtcaaga cttggcaaaa caagcattgc aagaaattgt
     1441 tattcttcct tctctgaggc ctgagttgtt cacagggctt agagctcctg ccagagggct
     1501 gttactcttt ggtccacctg ggaatgggaa gacaatgctg gctaaagcag tagctgcaga
     1561 atcgaatgca accttcttta atataagtgc tgcaagttta acttcaaaat acgtgggaga
     1621 aggagagaaa ttggtgaggg ctctttttgc tgtggctcga gaacttcaac cttctataat
     1681 ttttatagat gaagttgata gccttttgtg tgaaagaaga gaaggggagc acgatgctag
     1741 tagacgccta aaaactgaat ttctaataga atttgatggt gtacagtctg ctggagatga
     1801 cagagtactt gtaatgggtg caactaatag gccacaagag cttgatgagg ctgttctcag
     1861 gcgtttcatc aaacgggtat atgtgtcttt accaaatgag gagacaagac tacttttgct
     1921 taaaaatctg ttatgtaaac aaggaagtcc attgacccaa aaagaactag cacaacttgc
     1981 tagaatgact gatggatact caggaagtga cctaacagct ttggcaaaag atgcagcact
     2041 gggtcctatc cgagaactaa aaccagaaca ggtgaagaat atgtctgcca gtgagatgag
     2101 aaatattcga ttatctgact tcactgaatc cttgaaaaaa ataaaacgca gcgtcagccc
     2161 tcaaacttta gaagcgtaca tacgttggaa caaggacttt ggagatacca ctgtttaagg
     2221 aaataccttt gtaaacctgc agaacatttt acttaaaaga ggaaacacaa gatcctctag
     2281 tcagctgacg cgtgctagcg cggccgcatc gataagcttg tcgacgatat ctctagagga
     2341 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     2401 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     2461 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2521 cactgcctcg agcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2581 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2641 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2701 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2761 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2821 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2881 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2941 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3001 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3061 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3121 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3181 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3241 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3301 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3361 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3421 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3481 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3541 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3601 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3661 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3721 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3781 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3841 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     3901 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     3961 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4021 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4081 caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4141 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4201 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4261 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4321 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4381 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4441 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     4501 ataggcgtat cacgaggccc tttcgtcttc actc
//
