LOCUS       sequence-224743-        6034 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 6034)
  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 6034)
  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)-22Q-P80C
FEATURES             Location/Qualifiers
     source          1..6034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      15..101
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     236..255
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        236..254
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    255..279
                     /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             294..316
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             327..344
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1096..1114)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1110..1157
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1217..1321
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1322..1390,1391..2182)
                     /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(1540..1559)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2224..2737)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2419..2440)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2631..2650
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2848..3436
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3337..3356
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3590..3607
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3629..3648)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3789..3811
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3805..3996)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4553..5635)
                     /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     5597..5616
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5636..5713)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      5866..5952
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aattccccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
       61 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
      121 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccagga
      181 attggggatc ggaattaatt cccggtttaa accggggatc tcgatcccgc gaaattaata
      241 cgactcacta taggggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt
      301 aactttaaga aggagatata catatgcatc atcatcatca ccatagcggt gcaattagcg
      361 gtgatagcct gattagcctg gcaagcaccg gtaaacgtgt tagcattaaa gatctgctgg
      421 atgagaaaga ttttgagatt tgggccatta acgagcagac catgaaactg gaaagcgcaa
      481 aagttagccg tgttttttgc accggtaaaa aactggtgta tatcctgaaa acccgtctgg
      541 gtcgtaccat taaagcaacc gcaaatcatc gttttctgac cattgatggt tggaaacgtc
      601 tggatgaact gagcctgaaa gaacatattg cactgcctcg taaactggaa agtagcagcc
      661 tgcaactgag tccggaaatt gaaaaactga gccagagcga tatctattgg gatagcattg
      721 ttagcattac cgaaaccggt gttgaagaag tttttgatct gaccgttccg ggtccgcaca
      781 attttgttgc aaatgatatt atcgtgcata attgccagca gcagcaacaa cagcagcaac
      841 agcaacagca gcagcagcag caacaacaac agcagcaaca accgcctccg cctccgccac
      901 ctcctcctcc accgcagtta ccgcagcctc cgcctcaggc acagccgtta ctgccgcagc
      961 ctcagccacc gcctccacct ccccctccgc ctccaggttg tgcagttgca gaagaaccgc
     1021 tgcatcgtcc gtaactgcag gaaggggatc cggctgctaa caaagcccga aaggaagctg
     1081 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     1141 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg ataactacgt caggtggcac
     1201 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     1261 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     1321 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     1381 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     1441 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     1501 cgaagaacgt ttcccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     1561 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     1621 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     1681 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     1741 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     1801 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     1861 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     1921 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     1981 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     2041 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     2101 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     2161 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     2221 tttaccccgg ttgataatca gaaaagcccc aaaaacagga agattgtata agcaaatatt
     2281 taaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     2341 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagcccgag
     2401 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     2461 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     2521 aaatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2581 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2641 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2701 acacccgccg cgcttaatgc gccgctacag ggcgcgtaaa aggatctagg tgaagatcct
     2761 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2821 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2881 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2941 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     3001 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3061 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3121 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3181 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3241 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3301 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3361 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3421 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3481 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3541 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3601 gagcgaggaa gctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3661 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     3721 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3781 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     3841 gcagctgcgg taaagctcat cagcgtggtc gtgcagcgat tcacagatgt ctgcctgttc
     3901 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     3961 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     4021 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     4081 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     4141 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag ccgaacgcca gcaagacgta
     4201 gcccagcgcg tcggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt
     4261 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
     4321 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
     4381 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
     4441 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     4501 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     4561 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4621 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     4681 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     4741 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     4801 tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact
     4861 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     4921 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     4981 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
     5041 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     5101 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
     5161 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     5221 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     5281 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     5341 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     5401 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     5461 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     5521 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     5581 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
     5641 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     5701 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag
     5761 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgccggcat
     5821 gccgcccttt cgtcttcaag aattaattcc caattcccca ggcatcaaat aaaacgaaag
     5881 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     5941 agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg
     6001 cgggcaggac gcccgccata aactgccagg aatt
//
