LOCUS       sequence-222729-        6142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Htt fragment containing exon 1, with A2C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6142)
  AUTHORS   Warner JB 4th, Ruff KM, Tan PS, Lemke EA, Pappu RV, Lashuel HA
  TITLE     Monomeric Huntingtin Exon 1 Has Similar Overall Structural Features 
            for Wild-Type and Pathological Polyglutamine Lengths.
  JOURNAL   J Am Chem Soc. 2017 Oct 18;139(41):14456-14469. doi: 
            10.1021/jacs.7b06659. Epub 2017 Oct 9.
  PUBMED    28937758
REFERENCE   2  (bases 1 to 6142)
  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: 
            "10.1021/jacs.7b06659"; journalName: "J Am Chem Soc"; date: 
            "2017-10-18- 18"; volume: "139"; issue: "41"; pages: "14456-14469"
COMMENT     Sequence Label: pTWIN1-His6-Ssp-Httex1-43Q-A2C-P90C
FEATURES             Location/Qualifiers
     source          1..6142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      36..122
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     257..276
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        257..275
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    276..300
                     /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             315..337
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             348..365
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1225..1243)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1239..1286
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1346..1450
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1451..1519,1520..2311)
                     /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(1669..1688)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2353..2866)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2548..2569)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2760..2779
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2977..3565
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3466..3485
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3719..3736
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3758..3777)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3918..3940
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3934..4125)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4682..5764)
                     /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     5726..5745
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5765..5842)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      5995..6081
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cgccataaac tgccaggaat taattcccca ggcatcaaat aaaacgaaag gctcagtcga
       61 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
      121 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
      181 gcccgccata aactgccagg aattggggat cggaattaat tcccggttta aaccggggat
      241 ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
      301 cctctagaaa taattttgtt taactttaag aaggagatat acatatgcat catcatcatc
      361 accatagcgg tgcaattagc ggtgatagcc tgattagcct ggcaagcacc ggtaaacgtg
      421 ttagcattaa agatctgctg gatgagaaag attttgagat ttgggccatt aacgagcaga
      481 ccatgaaact ggaaagcgca aaagttagcc gtgttttttg caccggtaaa aaactggtgt
      541 atatcctgaa aacccgtctg ggtcgtacca ttaaagcaac cgcaaatcat cgttttctga
      601 ccattgatgg ttggaaacgt ctggatgaac tgagcctgaa agaacatatt gcactgcctc
      661 gtaaactgga aagtagcagc ctgcaactga gtccggaaat tgaaaaactg agccagagcg
      721 atatctattg ggatagcatt gttagcatta ccgaaaccgg tgttgaagaa gtttttgatc
      781 tgaccgttcc gggtccgcac aattttgttg caaatgatat tattgtgcac aactgcaccc
      841 tggaaaaact gatgaaagca tttgaaagcc tgaaaagctt tcagcagcag caacaacagc
      901 agcaacagca acagcagcag cagcagcaac aacagcagca gcaacagcaa caacaacaac
      961 aacagcagca acaacagcaa cagcagcaac aacaacagca acagcaacaa ccgcctccgc
     1021 ctccgccacc tcctcctcca ccacagctgc ctcagcctcc gcctcaggca cagcctctgc
     1081 tgccacagcc tcagccacct ccaccgcctc cacctccacc tccgggtccg gcagttgcag
     1141 aagaaccgct gcatcgttgc taactgcagg aaggggatcc ggctgctaac aaagcccgaa
     1201 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     1261 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga taactacgtc
     1321 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1381 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1441 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     1501 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     1561 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     1621 ttttcgcccc gaagaacgtt tcccaatgat gagcactttt aaagttctgc tatgtggcgc
     1681 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     1741 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     1801 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     1861 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     1921 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     1981 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2041 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2101 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2161 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2221 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     2281 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     2341 ttagattgat ttaccccggt tgataatcag aaaagcccca aaaacaggaa gattgtataa
     2401 gcaaatattt aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     2461 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     2521 tagcccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     2581 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     2641 ccatcaccca aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     2701 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     2761 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     2821 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtaaaa ggatctaggt
     2881 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2941 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3001 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3061 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3121 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3181 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3241 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3301 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3361 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3421 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3481 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3541 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3601 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3661 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3721 cgagtcagtg agcgaggaag ctatggtgca ctctcagtac aatctgctct gatgccgcat
     3781 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
     3841 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     3901 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     3961 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgcagcgatt cacagatgtc
     4021 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     4081 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta
     4141 agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat
     4201 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
     4261 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc cgaacgccag
     4321 caagacgtag cccagcgcgt cggccgccat gccggcgata atggcctgct tctcgccgaa
     4381 acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac
     4441 cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac
     4501 ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc
     4561 ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
     4621 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     4681 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4741 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     4801 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     4861 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     4921 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca
     4981 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     5041 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     5101 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     5161 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     5221 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     5281 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     5341 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     5401 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     5461 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     5521 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     5581 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     5641 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     5701 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     5761 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     5821 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     5881 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     5941 tgccggcatg ccgccctttc gtcttcaaga attaattccc aattccccag gcatcaaata
     6001 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     6061 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
     6121 ggagggtggc gggcaggacg cc
//
