LOCUS       sequence-222733-        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-P90C
FEATURES             Location/Qualifiers
     source          1..6034
                     /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(1111..1129)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1125..1172
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1232..1336
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1337..1405,1406..2197)
                     /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(1555..1574)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2239..2752)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2434..2455)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2646..2665
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2863..3451
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3352..3371
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3605..3622
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3644..3663)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3804..3826
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3820..4011)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4568..5650)
                     /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     5612..5631
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5651..5728)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      5881..5967
                     /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 atattatcgt gcataattgc cagcagcagc
      841 aacaacagca gcaacagcaa cagcagcagc agcagcaaca acaacagcag caacaaccgc
      901 ctccgcctcc gccacctcct cctccaccac agctgcctca gcctccgcct caggcacagc
      961 ctctgctgcc acagcctcag ccacctccac cgcctccacc tccacctccg ggtccggcag
     1021 ttgcagaaga accgctgcat cgttgttaac tgcaggaagg ggatccggct gctaacaaag
     1081 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     1141 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggataac
     1201 tacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1261 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1321 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1381 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     1441 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     1501 tgagagtttt cgccccgaag aacgtttccc aatgatgagc acttttaaag ttctgctatg
     1561 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     1621 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     1681 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     1741 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     1801 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     1861 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     1921 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     1981 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     2041 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     2101 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     2161 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2221 tatactttag attgatttac cccggttgat aatcagaaaa gccccaaaaa caggaagatt
     2281 gtataagcaa atatttaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt
     2341 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     2401 aaagaatagc ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     2461 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2521 cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2581 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     2641 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     2701 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtaaaaggat
     2761 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     2821 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2881 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2941 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3001 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3061 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3121 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3181 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3241 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3301 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3361 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3421 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3481 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3541 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3601 gcgcagcgag tcagtgagcg aggaagctat ggtgcactct cagtacaatc tgctctgatg
     3661 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     3721 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3781 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3841 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgca gcgattcaca
     3901 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     3961 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     4021 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     4081 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     4141 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagccgaa
     4201 cgccagcaag acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc
     4261 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     4321 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     4381 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     4441 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     4501 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     4561 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4621 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     4681 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     4741 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     4801 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa
     4861 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     4921 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     4981 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     5041 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     5101 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
     5161 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
     5221 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
     5281 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
     5341 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
     5401 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
     5461 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     5521 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     5581 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
     5641 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     5701 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
     5761 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     5821 ggtgcatgcc ggcatgccgc cctttcgtct tcaagaatta attcccaatt ccccaggcat
     5881 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     5941 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa
     6001 cggcccggag ggtggcgggc aggacgcccg ccat
//
