LOCUS       sequence-255885-        6481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacteiral expression of Spastin M87 varient without exon 4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6481)
  AUTHORS   Han H, Schubert HL, McCullough J, Monroe N, Purdy MD, Yeager M, 
            Sundquist WI, Hill CP
  TITLE     Structure of spastin bound to a glutamate-rich peptide implies a 
            hand-over-hand mechanism of substrate translocation.
  JOURNAL   J Biol Chem. 2019 Nov 25. pii: AC119.009890. doi: 
            10.1074/jbc.AC119.009890.
  PUBMED    31767681
REFERENCE   2  (bases 1 to 6481)
  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: "J Biol 
            Chem. 2019 Nov 25. pii: AC119.009890. doi: 
            10.1074/jbc.AC119.009890."
COMMENT     Sequence Label: pGEX-PP-Spastin(87-616)DeltaExon4
FEATURES             Location/Qualifiers
     source          1..6481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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)."
     CDS             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..935
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             939..962
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2698..2716)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2784..2888
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2889..2957,2958..3749)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3107..3126)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3920..4508
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4409..4428
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4662..4679
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4752..4829
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4830..5912
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4849..4868)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5961..5966,5967..5984,5985..5991)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5999..6015
                     /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)."
     primer_bind     6023..6039
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6023..6039
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6035..6481,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6055..6072)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6055..6071)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6064..6086)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gttccgcgtg gatctccgct tgaagtcctc tttcagggac
      961 cccatatggc agccaagagg agctccgggg ccgcgccagc acctgcctcg gcctcggccc
     1021 cggcgccggt gccgggcggc gaggccgagc gcgtccgagt cttccacaaa caggccttcg
     1081 agtacatctc cattgccctg cgcatcgatg aggatgagaa agcaggacag aaggagcaag
     1141 ctgtggaatg gtataagaaa ggtattgaag aactggaaaa aggaatagct gttatagtta
     1201 caggacaagg tgaacagtgt gaaagagcta gacgccttca agctaaaatg atgactaatt
     1261 tggttatggc caaggaccgc ttacaacttc tagagagtgg agctgttcca aaaagaaaag
     1321 accccttaac acacactagt aattcactgc ctcgttcaaa aacagtcatg aaaactggat
     1381 ctgcaggcct ttcaggccac catagagcac ctagttacag tggtttatcc atggtttctg
     1441 gagtgaaaca gggatctggt cctgctccta ccactcataa gggtactccg aaaacaaata
     1501 ggacaaataa accttctacc cctacaactg ctactcgtaa gaaaaaagac ttgaagaatt
     1561 ttaggaatgt ggacagcaac cttgctaacc ttataatgaa tgaaattgtg gacaatggaa
     1621 cagctgttaa atttgatgat atagctggtc aagacttggc aaaacaagca ttgcaagaaa
     1681 ttgttattct tccttctctg aggcctgagt tgttcacagg gcttagagct cctgccagag
     1741 ggctgttact ctttggtcca cctgggaatg ggaagacaat gctggctaaa gcagtagctg
     1801 cagaatcgaa tgcaaccttc tttaatataa gtgctgcaag tttaacttca aaatacgtgg
     1861 gagaaggaga gaaattggtg agggctcttt ttgctgtggc tcgagaactt caaccttcta
     1921 taatttttat agatgaagtt gatagccttt tgtgtgaaag aagagaaggg gagcacgatg
     1981 ctagtagacg cctaaaaact gaatttctaa tagaatttga tggtgtacag tctgctggag
     2041 atgacagagt acttgtaatg ggtgcaacta ataggccaca agagcttgat gaggctgttc
     2101 tcaggcgttt catcaaacgg gtatatgtgt ctttaccaaa tgaggagaca agactacttt
     2161 tgcttaaaaa tctgttatgt aaacaaggaa gtccattgac ccaaaaagaa ctagcacaac
     2221 ttgctagaat gactgatgga tactcaggaa gtgacctaac agctttggca aaagatgcag
     2281 cactgggtcc tatccgagaa ctaaaaccag aacaggtgaa gaatatgtct gccagtgaga
     2341 tgagaaatat tcgattatct gacttcactg aatccttgaa aaaaataaaa cgcagcgtca
     2401 gccctcaaac tttagaagcg tacatacgtt ggaacaagga ctttggagat accactgttt
     2461 aaggatcctc gagaattcat cgtgactgac tgacgatctg cctcgcgcgt ttcggtgatg
     2521 acggtgaaaa cctctgacac atgcggctcc cggagacggt cacagcttgt ctgtaagcgg
     2581 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg
     2641 cagccatgac ccagtcacgt agcgatagcg gagtgtataa ttcttgaaga cgaaagggcc
     2701 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     2761 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2821 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2881 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     2941 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3001 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3061 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3121 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3181 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3241 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3301 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatggga gatcatgtaa
     3361 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3421 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3481 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3541 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3601 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3661 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3721 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     3781 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     3841 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3901 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3961 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4021 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     4081 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4141 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4201 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4261 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4321 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4381 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4441 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4501 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4561 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     4621 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     4681 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4741 gcataaattc cgacaccatc gaatggcgca aaacctttcg cggtatggca tgatagcgcc
     4801 cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag
     4861 agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt
     4921 ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc
     4981 gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc
     5041 tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg
     5101 gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg
     5161 tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc
     5221 aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct
     5281 ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg
     5341 tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt
     5401 ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc
     5461 agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc
     5521 aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc
     5581 tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag
     5641 tgggatacga cgataccgaa gacagctcat gttatatccc gccgttaacc accatcaaac
     5701 aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc
     5761 aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa accaccctgg
     5821 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     5881 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt aagttagctc
     5941 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     6001 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgg attcactggc
     6061 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     6121 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     6181 ccaacagttg cgcagcctga atggcgaatg gcgctttgcc tggtttccgg caccagaagc
     6241 ggtgccggaa agctggctgg agtgcgatct tcctgaggcc gatactgtcg tcgtcccctc
     6301 aaactggcag atgcacggtt acgatgcgcc catctacacc aacgtgacct atcccattac
     6361 ggtcaatccg ccgtttgttc ccacggagaa tccgacgggt tgttactcgc tcacatttaa
     6421 tgttgatgaa agctggctac aggaaggcca gacgcgaatt atttttgatg gcgttggaat
     6481 t
//
