LOCUS       sequence-200846-        6061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the human Huntingtin Exon1 protein containing 15Q in 
            E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6061)
  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 6061)
  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-Httex1-15Q
FEATURES             Location/Qualifiers
     source          1..6061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      43..129
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     264..283
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        264..282
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    283..307
                     /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             322..344
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             355..372
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1151..1169)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1165..1212
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1272..1376
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1377..1445,1446..2237)
                     /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(1595..1614)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2279..2792)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2474..2495)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2686..2705
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2903..3491
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3392..3411
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3645..3662
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3684..3703)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3844..3866
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3860..4051)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4608..5690)
                     /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     5652..5671
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5691..5768)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      5921..6007
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggacgcccgc cataaactgc caggaattaa ttccccaggc atcaaataaa acgaaaggct
       61 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      121 aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg
      181 gcaggacgcc cgccataaac tgccaggaat tggggatcgg aattaattcc cggtttaaac
      241 cggggatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
      301 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgcatcat
      361 catcatcacc atagcggtgc aattagcggt gatagcctga ttagcctggc aagcaccggt
      421 aaacgtgtta gcattaaaga tctgctggat gagaaagatt ttgagatttg ggccattaac
      481 gagcagacca tgaaactgga aagcgcaaaa gttagccgtg ttttttgcac cggtaaaaaa
      541 ctggtgtata tcctgaaaac ccgtctgggt cgtaccatta aagcaaccgc aaatcatcgt
      601 tttctgacca ttgatggttg gaaacgtctg gatgaactga gcctgaaaga acatattgca
      661 ctgcctcgta aactggaaag tagcagcctg caactgagtc cggaaattga aaaactgagc
      721 cagagcgata tctattggga tagcattgtt agcattaccg aaaccggtgt tgaagaagtt
      781 tttgatctga ccgttccggg tccgcacaat tttgttgcaa atgatattat tgtgcacaac
      841 gccaccctgg aaaaactgat gaaagcattt gaaagcctga aaagctttca gcagcagcaa
      901 caacagcagc aacagcaaca acaacaacag cagcctccgc ctccgcctcc tccaccacct
      961 cctccacagc tgcctcagcc tccaccgcag gcacagcctc tgctgccaca gcctcagcca
     1021 cctccaccgc caccgcctcc acctccgggt ccagcagttg cagaagaacc gctgcatcgt
     1081 ccgtaataac tgcaggaagg ggatccggct gctaacaaag cccgaaagga agctgagttg
     1141 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     1201 aggggttttt tgctgaaagg aggaactata tccggataac tacgtcaggt ggcacttttc
     1261 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1321 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     1381 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1441 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     1501 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     1561 aacgtttccc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     1621 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     1681 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     1741 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     1801 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     1861 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     1921 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     1981 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2041 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     2101 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     2161 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     2221 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttac
     2281 cccggttgat aatcagaaaa gccccaaaaa caggaagatt gtataagcaa atatttaaat
     2341 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     2401 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaatagc ccgagatagg
     2461 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     2521 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat cacccaaatc
     2581 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     2641 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     2701 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     2761 cgccgcgctt aatgcgccgc tacagggcgc gtaaaaggat ctaggtgaag atcctttttg
     2821 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2881 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2941 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3001 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     3061 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3121 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3181 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3241 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3301 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3361 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3421 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3481 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3541 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3601 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3661 aggaagctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     3721 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     3781 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3841 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     3901 tgcggtaaag ctcatcagcg tggtcgtgca gcgattcaca gatgtctgcc tgttcatccg
     3961 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     4021 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     4081 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     4141 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     4201 gggaccagag aaaaatcact cagggtcaat gccagccgaa cgccagcaag acgtagccca
     4261 gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     4321 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     4381 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     4441 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     4501 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
     4561 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     4621 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4681 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     4741 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     4801 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     4861 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
     4921 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     4981 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     5041 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     5101 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     5161 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     5221 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     5281 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     5341 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     5401 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     5461 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     5521 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     5581 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     5641 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     5701 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     5761 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     5821 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgcc ggcatgccgc
     5881 cctttcgtct tcaagaatta attcccaatt ccccaggcat caaataaaac gaaaggctca
     5941 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     6001 gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc
     6061 a
//
