LOCUS       sequence-181063-        4740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of S245X spastin M1 and M87 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4740)
  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 4740)
  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 S245X with 5'UTR
FEATURES             Location/Qualifiers
     source          1..4740
                     /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    2651..2732
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2688..2707)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2743..2760)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2914..3502)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2994..3013)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3673..4464,4465..4533))
                     /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     4296..4315
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4534..4638)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4706..4724
                     /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 cggtacccga cccacgcgtc cgggcccgag
      361 ccaccgactg caggaggaga aggggttgtg ctcctggccg aggaaggaga aaggggcggg
      421 gccggcgggc agcgtgcggc agtgcggagc tcctgagacc ggcgggcaca cgggggtctg
      481 tggcccccgc cgtagcagtg gctgccgccg tcgcttggtt cccgtcggtc tgcgggaggc
      541 gggttatggc ggcggcggca gtgagagctg tgaatgaatt ctccgggtgg acgagggaag
      601 aagaaaggct ccggcggcgc cagcaacccg gtgcctccca ggcctccgcc cccttgcctg
      661 gcccccgccc ctcccgccgc cgggccggcc cctccgcccg agtcgccgca taagcggaac
      721 ctgtactatt tctcctaccc gctgtttgta ggcttcgcgc tgctgcgttt ggtcgccttc
      781 cacctggggc tcctcttcgt gtggctctgc cagcgcttct cccgcgccct catggcagcc
      841 aagaggagct ccggggccgc gccagcacct gcctcggcct cggccccggc gccggtgccg
      901 ggcggcgagg ccgagcgcgt ccgagtcttc cacaaacagg ccttcgagta catctccatt
      961 gccctgcgca tcgatgagga tgagaaagca ggacagaagg agcaagctgt ggaatggtat
     1021 aagaaaggta ttgaagaact ggaaaaagga atagctgtta tagttacagg acaaggtgaa
     1081 cagtgtgaaa gagctagacg ccttcaagct aaaatgatga ctaatttggt tatggccaag
     1141 gaccgcttac aacttctaga gaagatgcaa ccagttttgc cattttccaa gtcacaaacg
     1201 gacgtctata atgacagtac taacttggca tgccgcaatg gacatctcca gtcagaaagt
     1261 ggagctgttc caaaaagaaa agacccctta acacacacta gtaattgact gcctcgttca
     1321 aaaacagtta tgaaaactgg atctgcaggc ctttcaggcc accatagagc acctagttac
     1381 agtggtttat ccatggtttc tggagtgaaa cagggatctg gtcctgctcc taccactcat
     1441 aagggtactc cgaaaacaaa taggacaaat aaaccttcta cccctacaac tgctactcgt
     1501 aagaaaaaag acttgaagaa ttttaggaat gtggacagca accttgctaa ccttataatg
     1561 aatgaaattg tggacaatgg aacagctgtt aaatttgatg atatagctgg tcaagacttg
     1621 gcaaaacaag cattgcaaga aattgttatt cttccttctc tgaggcctga gttgttcaca
     1681 gggcttagag ctcctgccag agggctgtta ctctttggtc cacctgggaa tgggaagaca
     1741 atgctggcta aagcagtagc tgcagaatcg aatgcaacct tctttaatat aagtgctgca
     1801 agtttaactt caaaatacgt gggagaagga gagaaattgg tgagggctct ttttgctgtg
     1861 gctcgagaac ttcaaccttc tataattttt atagatgaag ttgatagcct tttgtgtgaa
     1921 agaagagaag gggagcacga tgctagtaga cgcctaaaaa ctgaatttct aatagaattt
     1981 gatggtgtac agtctgctgg agatgacaga gtacttgtaa tgggtgcaac taataggcca
     2041 caagagcttg atgaggctgt tctcaggcgt ttcatcaaac gggtatatgt gtctttacca
     2101 aatgaggaga caagactact tttgcttaaa aatctgttat gtaaacaagg aagtccattg
     2161 acccaaaaag aactagcaca acttgctaga atgactgatg gatactcagg aagtgaccta
     2221 acagctttgg caaaagatgc agcactgggt cctatccgag aactaaaacc agaacaggtg
     2281 aagaatatgt ctgccagtga gatgagaaat attcgattat ctgacttcac tgaatccttg
     2341 aaaaaaataa aacgcagcgt cagccctcaa actttagaag cgtacatacg ttggaacaag
     2401 gactttggag ataccactgt ttaaggaaat acctttgtaa acctgcagaa cattttactt
     2461 aaaagaggaa acacaagatc ctctagtcag ctgacgcgtg ctagcgcggc cgcatcgata
     2521 agcttgtcga cgatatctct agaggatcat aatcagccat accacatttg tagaggtttt
     2581 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     2641 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2701 aaatttcaca aataaagcat ttttttcact gcctcgagct tcctcgctca ctgactcgct
     2761 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2821 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2881 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2941 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3001 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3061 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3121 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3181 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3241 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3301 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3361 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3421 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3481 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3541 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3601 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3661 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3721 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3781 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3841 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3901 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3961 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4021 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4081 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4141 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4201 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4261 gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac
     4321 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4381 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4441 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4501 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4561 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4621 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     4681 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcactc
//
