LOCUS       sequence-222736-        6094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Htt fragment containing amino acids 18-90, 
            with Q18C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6094)
  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 6094)
  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-Htt18-90(Q18C)-42Q-A60C
FEATURES             Location/Qualifiers
     source          1..6094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      44..130
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     265..284
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        265..283
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    284..308
                     /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             323..345
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             356..373
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1185..1203)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1199..1246
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1306..1410
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1411..1479,1480..2271)
                     /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(1629..1648)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2313..2826)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2508..2529)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2720..2739
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2937..3525
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3426..3445
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3679..3696
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3718..3737)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3878..3900
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3894..4085)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4642..5724)
                     /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     5686..5705
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5725..5802)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      5955..6041
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aggacgcccg ccataaactg ccaggaatta attccccagg catcaaataa aacgaaaggc
       61 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
      121 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg
      181 ggcaggacgc ccgccataaa ctgccaggaa ttggggatcg gaattaattc ccggtttaaa
      241 ccggggatct cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata
      301 acaattcccc tctagaaata attttgttta actttaagaa ggagatatac atatgcatca
      361 tcatcatcac catagcggtg caattagcgg tgatagcctg attagcctgg caagcaccgg
      421 taaacgtgtt agcattaaag atctgctgga tgagaaagat tttgagattt gggccattaa
      481 cgagcagacc atgaaactgg aaagcgcaaa agttagccgt gttttttgca ccggtaaaaa
      541 actggtgtat atcctgaaaa cccgtctggg tcgtaccatt aaagcaaccg caaatcatcg
      601 ttttctgacc attgatggtt ggaaacgtct ggatgaactg agcctgaaag aacatattgc
      661 actgcctcgt aaactggaaa gtagcagcct gcaactgagt ccggaaattg aaaaactgag
      721 ccagagcgat atctattggg atagcattgt tagcattacc gaaaccggtg ttgaagaagt
      781 ttttgatctg accgttccgg gtccgcacaa ttttgttgca aatgatatta tcgtgcataa
      841 ttgccagcag cagcaacaac agcagcaaca gcaacagcag cagcagcagc aacaacagca
      901 gcagcaacag caacaacaac aacaacagca gcaacaacag caacagcagc aacaacaaca
      961 gcaacagcag cctccgcctc cgccacctcc tcctccacca cctcagctgc ctcagcctcc
     1021 gcctcagtgc cagccgctgc tgccacagcc tcagccaccg cctccaccgc caccacctcc
     1081 gcctggtcca gcagttgcag aagaaccgct gcatcgtccg taactgcagg aaggggatcc
     1141 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     1201 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     1261 tatatccgga taactacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     1321 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1381 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     1441 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     1501 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     1561 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt tcccaatgat gagcactttt
     1621 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     1681 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1741 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     1801 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     1861 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     1921 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     1981 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     2041 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     2101 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     2161 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     2221 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     2281 caagtttact catatatact ttagattgat ttaccccggt tgataatcag aaaagcccca
     2341 aaaacaggaa gattgtataa gcaaatattt aaattgtaaa cgttaatatt ttgttaaaat
     2401 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     2461 tcccttataa atcaaaagaa tagcccgaga tagggttgag tgttgttcca gtttggaaca
     2521 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     2581 gcgatggccc actacgtgaa ccatcaccca aatcaagttt tttggggtcg aggtgccgta
     2641 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     2701 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     2761 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     2821 gcgcgtaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     2881 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2941 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3001 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3061 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     3121 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3181 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3241 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3301 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3361 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3421 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3481 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3541 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3601 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3661 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag ctatggtgca ctctcagtac
     3721 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     3781 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     3841 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     3901 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     3961 tgcagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     4021 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     4081 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     4141 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     4201 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     4261 caatgccagc cgaacgccag caagacgtag cccagcgcgt cggccgccat gccggcgata
     4321 atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg
     4381 gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag
     4441 cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata
     4501 aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg
     4561 actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa
     4621 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4681 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
     4741 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
     4801 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
     4861 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
     4921 gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
     4981 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
     5041 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
     5101 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
     5161 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
     5221 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
     5281 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
     5341 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
     5401 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
     5461 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
     5521 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
     5581 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
     5641 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
     5701 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
     5761 tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc
     5821 ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg
     5881 ccgccgcaag gaatggtgca tgccggcatg ccgccctttc gtcttcaaga attaattccc
     5941 aattccccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     6001 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     6061 acgttgcgaa gcaacggccc ggagggtggc gggc
//
