LOCUS       sequence-202356-        6082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the human Huntingtin Exon1 protein containing 23Q in 
            E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6082)
  AUTHORS   Vieweg S, Ansaloni A, Wang ZM, Warner JB, Lashuel HA
  TITLE     An Intein-based Strategy for the Production of Tag-free Huntingtin 
            Exon 1 Proteins Enables New Insights into the Polyglutamine 
            Dependence of Httex1 Aggregation and Fibril Formation.
  JOURNAL   J Biol Chem. 2016 Jun 3;291(23):12074-86. doi: 
            10.1074/jbc.M116.713982. Epub 2016 Mar 21.
  PUBMED    27002149
REFERENCE   2  (bases 1 to 6082)
  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"; doi: 
            "10.1074/jbc.M116.713982"; journalName: "J Biol Chem"; date: 
            "2016-06-3- 3"; volume: "291"; issue: "23"; pages: "12074-86"
COMMENT     Sequence Label: pTWIN1-His6-Ssp-Httex1-23Q
FEATURES             Location/Qualifiers
     source          1..6082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      38..124
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      221..307
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     442..461
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        442..460
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    461..485
                     /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)."
     RBS             500..522
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             533..550
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1350..1368)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1364..1411
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1471..1575
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1576..1644,1645..2436)
                     /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     complement(1794..1813)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2478..2991)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2673..2694)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2885..2904
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      3102..3690
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3591..3610
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3844..3861
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3883..3902)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4043..4065
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4059..4250)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4807..5889)
                     /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     5851..5870
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5890..5967)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ttcgtcttca agaattaatt cccaattccc caggcatcaa ataaaacgaa aggctcagtc
       61 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
      121 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg
      181 acgcccgcca taaactgcca ggaattaatt ccccaggcat caaataaaac gaaaggctca
      241 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
      301 gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc
      361 aggacgcccg ccataaactg ccaggaattg gggatcggaa ttaattcccg gtttaaaccg
      421 gggatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca
      481 attcccctct agaaataatt ttgtttaact ttaagaagga gatatacata tgcatcatca
      541 tcatcaccat agcggtgcaa ttagcggtga tagcctgatt agcctggcaa gcaccggtaa
      601 acgtgttagc attaaagatc tgctggatga gaaagatttt gagatttggg ccattaacga
      661 gcagaccatg aaactggaaa gcgcaaaagt tagccgtgtt ttttgcaccg gtaaaaaact
      721 ggtgtatatc ctgaaaaccc gtctgggtcg taccattaaa gcaaccgcaa atcatcgttt
      781 tctgaccatt gatggttgga aacgtctgga tgaactgagc ctgaaagaac atattgcact
      841 gcctcgtaaa ctggaaagta gcagcctgca actgagtccg gaaattgaaa aactgagcca
      901 gagcgatatc tattgggata gcattgttag cattaccgaa accggtgttg aagaagtttt
      961 tgatctgacc gttccgggtc cgcacaattt tgttgcaaat gatattatcg tgcataatgc
     1021 caccctggaa aaactgatga aagcatttga aagcctgaaa agctttcagc aacaacagca
     1081 gcaacagcaa cagcagcagc agcagcaaca acagcagcag caacagcaac aacaaccgcc
     1141 tccgcctccg ccacctcctc ctccaccaca gctgcctcag cctccgcctc aggcacagcc
     1201 tctgctgcca cagcctcagc cacctccacc gcctccacct ccacctccgg gtccggcagt
     1261 tgcagaagaa ccgctgcatc gtccgtaact gcaggaaggg gatccggctg ctaacaaagc
     1321 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     1381 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggataact
     1441 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1501 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     1561 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     1621 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     1681 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     1741 gagagttttc gccccgaaga acgtttccca atgatgagca cttttaaagt tctgctatgt
     1801 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     1861 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     1921 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     1981 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     2041 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     2101 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     2161 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     2221 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     2281 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     2341 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     2401 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     2461 atactttaga ttgatttacc ccggttgata atcagaaaag ccccaaaaac aggaagattg
     2521 tataagcaaa tatttaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt
     2581 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     2641 aagaatagcc cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2701 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2761 gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     2821 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     2881 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     2941 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg taaaaggatc
     3001 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3061 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3121 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3181 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3241 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3301 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3361 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3421 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3481 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3541 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3601 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3661 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3721 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3781 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3841 cgcagcgagt cagtgagcga ggaagctatg gtgcactctc agtacaatct gctctgatgc
     3901 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     3961 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4021 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4081 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgcag cgattcacag
     4141 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
     4201 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
     4261 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
     4321 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
     4381 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagccgaac
     4441 gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg
     4501 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
     4561 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
     4621 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
     4681 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     4741 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
     4801 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4861 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
     4921 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
     4981 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
     5041 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac
     5101 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
     5161 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
     5221 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
     5281 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
     5341 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
     5401 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
     5461 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
     5521 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
     5581 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
     5641 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
     5701 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
     5761 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
     5821 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
     5881 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
     5941 aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct
     6001 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
     6061 gtgcatgccg gcatgccgcc ct
//
