LOCUS       sequence-222664-        6145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Htt fragment containing exon 1, with A2C 
            cysteine mutation and a 43 polyQ repeat.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6145)
  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 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: 
            "10.1021/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
FEATURES             Location/Qualifiers
     source          1..6145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     114..133
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        114..132
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    133..157
                     /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             172..194
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             205..222
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1085..1103)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1099..1146
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1206..1310
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1311..1379,1380..2171)
                     /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(1529..1548)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2213..2726)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2408..2429)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2620..2639
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2837..3425
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3326..3345
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3579..3596
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3618..3637)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3778..3800
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3794..3985)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4542..5624)
                     /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     5586..5605
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5625..5702)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      5855..5941
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6038..6124
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggaat
       61 tggggatcgg aattaattcc cggtttaaac cggggatctc gatcccgcga aattaatacg
      121 actcactata ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa
      181 ctttaagaag gagatataca tatgcatcat catcatcacc atagcggtgc aattagcggt
      241 gatagcctga ttagcctggc aagcaccggt aaacgtgtta gcattaaaga tctgctggat
      301 gagaaagatt ttgagatttg ggccattaac gagcagacca tgaaactgga aagcgcaaaa
      361 gttagccgtg ttttttgcac cggtaaaaaa ctggtgtata tcctgaaaac ccgtctgggt
      421 cgtaccatta aagcaaccgc aaatcatcgt tttctgacca ttgatggttg gaaacgtctg
      481 gatgaactga gcctgaaaga acatattgca ctgcctcgta aactggaaag tagcagcctg
      541 caactgagtc cggaaattga aaaactgagc cagagcgata tctattggga tagcattgtt
      601 agcattaccg aaaccggtgt tgaagaagtt tttgatctga ccgttccggg tccgcacaat
      661 tttgttgcaa atgatattat tgtgcacaac tgcaccctgg aaaaactgat gaaagcattt
      721 gaaagcctga aaagctttca gcagcagcaa caacagcagc aacagcaaca gcagcagcag
      781 cagcaacaac agcagcagca acagcaacaa caacaacaac agcagcaaca acagcaacag
      841 cagcaacaac aacagcaaca gcaacaaccg cctccgcctc cgccacctcc tcctccacca
      901 cagctgcctc agcctccgcc tcaggcacag cctctgctgc cacagcctca gccacctcca
      961 ccgcctccac ctccacctcc gggtccggca gttgcagaag aaccgctgca tcgtccgtaa
     1021 taactgcagg aaggggatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     1081 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1141 tttttgctga aaggaggaac tatatccgga taactacgtc aggtggcact tttcggggaa
     1201 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1261 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     1321 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     1381 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     1441 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     1501 tcccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     1561 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     1621 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     1681 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     1741 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     1801 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     1861 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     1921 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     1981 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2041 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2101 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2161 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaccccggt
     2221 tgataatcag aaaagcccca aaaacaggaa gattgtataa gcaaatattt aaattgtaaa
     2281 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     2341 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga tagggttgag
     2401 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     2461 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca aatcaagttt
     2521 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     2581 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     2641 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     2701 gcttaatgcg ccgctacagg gcgcgtaaaa ggatctaggt gaagatcctt tttgataatc
     2761 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     2821 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     2881 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     2941 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     3001 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3061 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3121 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3181 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3241 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3301 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3361 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3421 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3481 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     3541 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     3601 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     3661 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     3721 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     3781 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     3841 aaagctcatc agcgtggtcg tgcagcgatt cacagatgtc tgcctgttca tccgcgtcca
     3901 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     3961 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     4021 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     4081 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     4141 agagaaaaat cactcagggt caatgccagc cgaacgccag caagacgtag cccagcgcgt
     4201 cggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     4261 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     4321 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     4381 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     4441 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     4501 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     4561 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4621 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     4681 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     4741 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     4801 tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg
     4861 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     4921 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     4981 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     5041 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     5101 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     5161 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     5221 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     5281 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     5341 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     5401 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     5461 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     5521 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     5581 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     5641 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     5701 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
     5761 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgccggcatg ccgccctttc
     5821 gtcttcaaga attaattccc aattccccag gcatcaaata aaacgaaagg ctcagtcgaa
     5881 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
     5941 tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg
     6001 cccgccataa actgccagga attaattccc caggcatcaa ataaaacgaa aggctcagtc
     6061 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     6121 aaatccgccg ggagcggatt tgaac
//
