LOCUS       Exported                4741 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Inducible expression of N184X spastin M1 and M87 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pTRE/SP N184X with 5'UTR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4741)
  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 4741)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4741
                     /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    2652..2733
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2689..2708)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2744..2761)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2915..3503)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2995..3014)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3674..4534)
                     /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     4297..4316
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4535..4639)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4707..4725
                     /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 cttaatttgg ttatggccaa
     1141 ggaccgctta caacttctag agaagatgca accagttttg ccattttcca agtcacaaac
     1201 ggacgtctat aatgacagta ctaacttggc atgccgcaat ggacatctcc agtcagaaag
     1261 tggagctgtt ccaaaaagaa aagacccctt aacacacact agtaattcac tgcctcgttc
     1321 aaaaacagtt atgaaaactg gatctgcagg cctttcaggc caccatagag cacctagtta
     1381 cagtggttta tccatggttt ctggagtgaa acagggatct ggtcctgctc ctaccactca
     1441 taagggtact ccgaaaacaa ataggacaaa taaaccttct acccctacaa ctgctactcg
     1501 taagaaaaaa gacttgaaga attttaggaa tgtggacagc aaccttgcta accttataat
     1561 gaatgaaatt gtggacaatg gaacagctgt taaatttgat gatatagctg gtcaagactt
     1621 ggcaaaacaa gcattgcaag aaattgttat tcttccttct ctgaggcctg agttgttcac
     1681 agggcttaga gctcctgcca gagggctgtt actctttggt ccacctggga atgggaagac
     1741 aatgctggct aaagcagtag ctgcagaatc gaatgcaacc ttctttaata taagtgctgc
     1801 aagtttaact tcaaaatacg tgggagaagg agagaaattg gtgagggctc tttttgctgt
     1861 ggctcgagaa cttcaacctt ctataatttt tatagatgaa gttgatagcc ttttgtgtga
     1921 aagaagagaa ggggagcacg atgctagtag acgcctaaaa actgaatttc taatagaatt
     1981 tgatggtgta cagtctgctg gagatgacag agtacttgta atgggtgcaa ctaataggcc
     2041 acaagagctt gatgaggctg ttctcaggcg tttcatcaaa cgggtatatg tgtctttacc
     2101 aaatgaggag acaagactac ttttgcttaa aaatctgtta tgtaaacaag gaagtccatt
     2161 gacccaaaaa gaactagcac aacttgctag aatgactgat ggatactcag gaagtgacct
     2221 aacagctttg gcaaaagatg cagcactggg tcctatccga gaactaaaac cagaacaggt
     2281 gaagaatatg tctgccagtg agatgagaaa tattcgatta tctgacttca ctgaatcctt
     2341 gaaaaaaata aaacgcagcg tcagccctca aactttagaa gcgtacatac gttggaacaa
     2401 ggactttgga gataccactg tttaaggaaa tacctttgta aacctgcaga acattttact
     2461 taaaagagga aacacaagat cctctagtca gctgacgcgt gctagcgcgg ccgcatcgat
     2521 aagcttgtcg acgatatctc tagaggatca taatcagcca taccacattt gtagaggttt
     2581 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     2641 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     2701 caaatttcac aaataaagca tttttttcac tgcctcgagc ttcctcgctc actgactcgc
     2761 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2821 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2881 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2941 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3001 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3061 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3121 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3181 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3241 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3301 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3361 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3421 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3481 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3541 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3601 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3661 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3721 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3781 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3841 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3901 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3961 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4021 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4081 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4141 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4201 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4261 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     4321 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4381 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4441 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4501 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4561 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4621 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     4681 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcact
     4741 c
//
