LOCUS       sequence-222716-        6409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Htt fragment containing amino acids 2-90, 
            with A2C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6409)
  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 6409)
  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-Htt2-90(A2C)-49Q-P90C
FEATURES             Location/Qualifiers
     source          1..6409
                     /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"
     terminator      198..284
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     419..438
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        419..437
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    438..462
                     /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             477..499
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             510..527
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1471..1489)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1485..1532
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1592..1696
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1697..1765,1766..2557)
                     /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(1915..1934)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2599..3112)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2794..2815)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3006..3025
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      3223..3811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3712..3731
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3965..3982
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4004..4023)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4164..4186
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4180..4371)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4928..6010)
                     /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     5972..5991
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6011..6088)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      6241..6327
                     /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 attaattccc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
      241 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
      301 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
      361 ggaattgggg atcggaatta attcccggtt taaaccgggg atctcgatcc cgcgaaatta
      421 atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga aataattttg
      481 tttaacttta agaaggagat atacatatgc atcatcatca tcaccatagc ggtgcaatta
      541 gcggtgatag cctgattagc ctggcaagca ccggtaaacg tgttagcatt aaagatctgc
      601 tggatgagaa agattttgag atttgggcca ttaacgagca gaccatgaaa ctggaaagcg
      661 caaaagttag ccgtgttttt tgcaccggta aaaaactggt gtatatcctg aaaacccgtc
      721 tgggtcgtac cattaaagca accgcaaatc atcgttttct gaccattgat ggttggaaac
      781 gtctggatga actgagcctg aaagaacata ttgcactgcc tcgtaaactg gaaagtagca
      841 gcctgcaact gagtccggaa attgaaaaac tgagccagag cgatatctat tgggatagca
      901 ttgttagcat taccgaaacc ggtgttgaag aagtttttga tctgaccgtt ccgggtccgc
      961 acaattttgt tgcaaatgat attattgtgc acaactgcac cctggaaaaa ctgatgaaag
     1021 catttgaaag cctgaaaagc tttcagcagc agcaacaaca gcagcaacag caacagcagc
     1081 agcagcagca acaacagcag cagcaacagc aacaacaaca acaacagcag caacaacagc
     1141 aacagcagca acaacaacag caacagcagc agcagcaaca acagcaacag cctccgcctc
     1201 cgccaccgcc tcctccacct ccgcagctgc cgcaacctcc tcctcaggca cagccgctgc
     1261 tgccgcagcc tcaacctcct ccaccacctc cacctccacc gcctggtccg gcagttgcag
     1321 aagaaccgct gcatcgttgc taactgcagg aagggaaccg ctgcatcgtt gctaactgca
     1381 ggaagggaga accgctgcat cgttgctaac tgcaggaagg ggatccggct gctaacaaag
     1441 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     1501 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggataac
     1561 tacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1621 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1681 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1741 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     1801 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     1861 tgagagtttt cgccccgaag aacgtttccc aatgatgagc acttttaaag ttctgctatg
     1921 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     1981 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2041 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     2101 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     2161 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     2221 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     2281 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     2341 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     2401 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     2461 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     2521 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2581 tatactttag attgatttac cccggttgat aatcagaaaa gccccaaaaa caggaagatt
     2641 gtataagcaa atatttaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt
     2701 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     2761 aaagaatagc ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     2821 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2881 cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2941 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3001 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3061 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtaaaaggat
     3121 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     3181 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3241 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3301 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3361 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3421 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3481 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3541 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3601 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3661 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3721 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3781 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3841 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3901 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3961 gcgcagcgag tcagtgagcg aggaagctat ggtgcactct cagtacaatc tgctctgatg
     4021 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     4081 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4141 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4201 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgca gcgattcaca
     4261 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     4321 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     4381 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     4441 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     4501 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagccgaa
     4561 cgccagcaag acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc
     4621 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     4681 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     4741 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     4801 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     4861 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     4921 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4981 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     5041 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     5101 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     5161 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa
     5221 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     5281 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     5341 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     5401 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     5461 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
     5521 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
     5581 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
     5641 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
     5701 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
     5761 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
     5821 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     5881 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     5941 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
     6001 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     6061 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
     6121 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     6181 ggtgcatgcc ggcatgccgc cctttcgtct tcaagaatta attcccaatt ccccaggcat
     6241 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     6301 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa
     6361 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggaatt
//
