LOCUS       Exported                4533 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Inducible expression of M1C448Y spastin in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pTRE/SP M1C448Y
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4533)
  AUTHORS   Solowska JM, D'Rozario M, Jean DC, Davidson MW, Marenda DR, Baas PW
  TITLE     Pathogenic mutation of spastin has gain-of-function effects on 
            microtubule dynamics.
  JOURNAL   J Neurosci. 2014 Jan 29;34(5):1856-67. doi: 
            10.1523/JNEUROSCI.3309-13.2014.
  PUBMED    24478365
REFERENCE   2  (bases 1 to 4533)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4533
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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    2444..2525
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2481..2500)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2536..2553)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2707..3295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2787..2806)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3466..4326)
                     /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     4089..4108
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4327..4431)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4499..4517
                     /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 tgactaattt ggttatggcc aaggaccgct tacaacttct agagaagatg
      961 caaccagttt tgccattttc caagtcacaa acggacgtct ataatgacag tactaacttg
     1021 gcatgccgca atggacatct ccagtcagaa agtggagctg ttccaaaaag aaaagacccc
     1081 ttaacacaca ctagtaattc actgcctcgt tcaaaaacag ttatgaaaac tggatctgca
     1141 ggcctttcag gccaccatag agcacctagt tacagtggtt tatccatggt ttctggagtg
     1201 aaacagggat ctggtcctgc tcctaccact cataagggta ctccgaaaac aaataggaca
     1261 aataaacctt ctacccctac aactgctact cgtaagaaaa aagacttgaa gaattttagg
     1321 aatgtggaca gcaaccttgc taaccttata atgaatgaaa ttgtggacaa tggaacagct
     1381 gttaaatttg atgatatagc tggtcaagac ttggcaaaac aagcattgca agaaattgtt
     1441 attcttcctt ctctgaggcc tgagttgttc acagggctta gagctcctgc cagagggctg
     1501 ttactctttg gtccacctgg gaatgggaag acaatgctgg ctaaagcagt agctgcagaa
     1561 tcgaatgcaa ccttctttaa tataagtgct gcaagtttaa cttcaaaata cgtgggagaa
     1621 ggagagaaat tggtgagggc tctttttgct gtggctcgag aacttcaacc ttctataatt
     1681 tttatagatg aagttgatag ccttttgtat gaaagaagag aaggggagca cgatgctagt
     1741 agacgcctaa aaactgaatt tctaatagaa tttgatggtg tacagtctgc tggagatgac
     1801 agagtacttg taatgggtgc aactaatagg ccacaagagc ttgatgaggc tgttctcagg
     1861 cgtttcatca aacgggtata tgtgtcttta ccaaatgagg agacaagact acttttgctt
     1921 aaaaatctgt tatgtaaaca aggaagtcca ttgacccaaa aagaactagc acaacttgct
     1981 agaatgactg atggatactc aggaagtgac ctaacagctt tggcaaaaga tgcagcactg
     2041 ggtcctatcc gagaactaaa accagaacag gtgaagaata tgtctgccag tgagatgaga
     2101 aatattcgat tatctgactt cactgaatcc ttgaaaaaaa taaaacgcag cgtcagccct
     2161 caaactttag aagcgtacat acgttggaac aaggactttg gagataccac tgtttaagga
     2221 aatacctttg taaacctgca gaacatttta cttaaaagag gaaacacaag atcctctagt
     2281 cagctgacgc gtgctagcgc ggccgcatcg ataagcttgt cgacgatatc tctagaggat
     2341 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     2401 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     2461 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2521 actgcctcga gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2581 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2641 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2701 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2761 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2821 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2881 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2941 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3001 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3061 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3121 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3181 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3241 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3301 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3361 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3421 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3481 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3541 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3601 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3661 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3721 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3781 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3841 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3901 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3961 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4021 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4081 aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4141 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4201 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4261 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4321 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4381 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4441 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     4501 taggcgtatc acgaggccct ttcgtcttca ctc
//
