LOCUS       sequence-200849-        6124 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the human Huntingtin Exon1 protein containing 37Q in 
            E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6124)
  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 6124)
  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-Httex1-37Q
FEATURES             Location/Qualifiers
     source          1..6124
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      30..116
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     251..270
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        251..269
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    270..294
                     /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             309..331
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             342..359
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1201..1219)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1215..1262
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1322..1426
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1427..1495,1496..2287)
                     /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(1645..1664)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2329..2842)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2524..2545)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2736..2755
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2953..3541
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3442..3461
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3695..3712
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3734..3753)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3894..3916
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3910..4101)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4658..5740)
                     /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     5702..5721
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5741..5818)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      5971..6057
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aaactgccag gaattaattc cccaggcatc aaataaaacg aaaggctcag tcgaaagact
       61 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
      121 cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc
      181 cataaactgc caggaattgg ggatcggaat taattcccgg tttaaaccgg ggatctcgat
      241 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta
      301 gaaataattt tgtttaactt taagaaggag atatacatat gcatcatcat catcaccata
      361 gcggtgcaat tagcggtgat agcctgatta gcctggcaag caccggtaaa cgtgttagca
      421 ttaaagatct gctggatgag aaagattttg agatttgggc cattaacgag cagaccatga
      481 aactggaaag cgcaaaagtt agccgtgttt tttgcaccgg taaaaaactg gtgtatatcc
      541 tgaaaacccg tctgggtcgt accattaaag caaccgcaaa tcatcgtttt ctgaccattg
      601 atggttggaa acgtctggat gaactgagcc tgaaagaaca tattgcactg cctcgtaaac
      661 tggaaagtag cagcctgcaa ctgagtccgg aaattgaaaa actgagccag agcgatatct
      721 attgggatag cattgttagc attaccgaaa ccggtgttga agaagttttt gatctgaccg
      781 ttccgggtcc gcacaatttt gttgcaaatg atattattgt gcacaacgcc accctggaaa
      841 aactgatgaa agcatttgaa agcctgaaaa gctttcagca gcagcaacaa cagcagcaac
      901 agcaacagca gcagcagcag caacaacagc agcagcaaca gcaacaacaa caacaacagc
      961 agcaacaaca gcaacagcag cagcaaccgc ctccgcctcc gccacctcct cctccaccac
     1021 agctgcctca gcctccgcct caggcacagc ctctgctgcc acagcctcag ccacctccac
     1081 cgcctccacc tccacctccg ggtccggcag ttgcagaaga accgctgcat cgtccgtaac
     1141 tgcaggaagg ggatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     1201 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     1261 tgctgaaagg aggaactata tccggataac tacgtcaggt ggcacttttc ggggaaatgt
     1321 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1381 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     1441 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     1501 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     1561 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgtttccc
     1621 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     1681 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     1741 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     1801 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1861 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1921 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1981 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2041 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2101 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     2161 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     2221 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2281 ttggtaactg tcagaccaag tttactcata tatactttag attgatttac cccggttgat
     2341 aatcagaaaa gccccaaaaa caggaagatt gtataagcaa atatttaaat tgtaaacgtt
     2401 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2461 gccgaaatcg gcaaaatccc ttataaatca aaagaatagc ccgagatagg gttgagtgtt
     2521 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2581 aaaaccgtct atcagggcga tggcccacta cgtgaaccat cacccaaatc aagttttttg
     2641 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2701 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2761 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2821 aatgcgccgc tacagggcgc gtaaaaggat ctaggtgaag atcctttttg ataatctcat
     2881 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2941 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3001 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3061 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     3121 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3181 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3241 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3301 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3361 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3421 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3481 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3541 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3601 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     3661 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagctat
     3721 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     3781 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     3841 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3901 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     3961 ctcatcagcg tggtcgtgca gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     4021 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     4081 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     4141 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     4201 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     4261 aaaaatcact cagggtcaat gccagccgaa cgccagcaag acgtagccca gcgcgtcggc
     4321 cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac
     4381 gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt
     4441 cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc
     4501 tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc
     4561 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc
     4621 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4681 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4741 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct
     4801 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc
     4861 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt
     4921 cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca
     4981 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat
     5041 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg
     5101 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg
     5161 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca
     5221 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg
     5281 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa
     5341 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa
     5401 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca
     5461 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt
     5521 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt
     5581 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg
     5641 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac
     5701 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac
     5761 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg
     5821 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg
     5881 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgcc ggcatgccgc cctttcgtct
     5941 tcaagaatta attcccaatt ccccaggcat caaataaaac gaaaggctca gtcgaaagac
     6001 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     6061 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
     6121 ccat
//
