LOCUS       sequence-200848-        6145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the human Huntingtin Exon1 protein containing 43Q in 
            E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6145)
  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 6145)
  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-Httex1-43Q
FEATURES             Location/Qualifiers
     source          1..6145
                     /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(1222..1240)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1236..1283
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1343..1447
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1448..1516,1517..2308)
                     /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(1666..1685)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2350..2863)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2545..2566)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2757..2776
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2974..3562
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3463..3482
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3716..3733
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3755..3774)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3915..3937
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3931..4122)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4679..5761)
                     /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     5723..5742
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5762..5839)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      5992..6078
                     /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 caacaacaac agcaacagca acaaccgcct ccgcctccgc
     1021 cacctcctcc tccaccacag ctgcctcagc ctccgcctca ggcacagcct ctgctgccac
     1081 agcctcagcc acctccaccg cctccacctc cacctccggg tccggcagtt gcagaagaac
     1141 cgctgcatcg tccgtaataa ctgcaggaag gggatccggc tgctaacaaa gcccgaaagg
     1201 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     1261 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggataa ctacgtcagg
     1321 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1381 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1441 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1501 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1561 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1621 tcgccccgaa gaacgtttcc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     1681 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1741 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     1801 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1861 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1921 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1981 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     2041 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2101 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2161 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2221 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2281 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2341 gattgattta ccccggttga taatcagaaa agccccaaaa acaggaagat tgtataagca
     2401 aatatttaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     2461 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     2521 cccgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     2581 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     2641 tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     2701 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     2761 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     2821 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtaaaagga tctaggtgaa
     2881 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     2941 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3001 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3061 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3121 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3181 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3241 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3301 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3361 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3421 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3481 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3541 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3601 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3661 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3721 gtcagtgagc gaggaagcta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     3781 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
     3841 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3901 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3961 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtgc agcgattcac agatgtctgc
     4021 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     4081 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg
     4141 gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg
     4201 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt
     4261 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagccga acgccagcaa
     4321 gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     4381 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     4441 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     4501 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     4561 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     4621 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     4681 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4741 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     4801 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     4861 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     4921 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc
     4981 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     5041 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     5101 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     5161 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     5221 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     5281 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     5341 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     5401 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     5461 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     5521 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     5581 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     5641 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     5701 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     5761 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     5821 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     5881 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
     5941 cggcatgccg ccctttcgtc ttcaagaatt aattcccaat tccccaggca tcaaataaaa
     6001 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     6061 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
     6121 gggtggcggg caggacgccc gccat
//
