LOCUS       sequence-222719-        6145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Htt fragment containing exon 1, with A2C.
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-A60C
FEATURES             Location/Qualifiers
     source          1..6145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      38..124
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     259..278
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        259..277
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    278..302
                     /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             317..339
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             350..367
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1230..1248)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1244..1291
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1351..1455
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1456..1524,1525..2316)
                     /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(1674..1693)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2358..2871)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2553..2574)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2765..2784
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2982..3570
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3471..3490
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3724..3741
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3763..3782)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3923..3945
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3939..4130)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4687..5769)
                     /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     5731..5750
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5770..5847)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      6000..6086
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cccgccataa actgccagga attaattccc caggcatcaa ataaaacgaa aggctcagtc
       61 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
      121 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg
      181 acgcccgcca taaactgcca ggaattgggg atcggaatta attcccggtt taaaccgggg
      241 atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt
      301 cccctctaga aataattttg tttaacttta agaaggagat atacatatgc atcatcatca
      361 tcaccatagc ggtgcaatta gcggtgatag cctgattagc ctggcaagca ccggtaaacg
      421 tgttagcatt aaagatctgc tggatgagaa agattttgag atttgggcca ttaacgagca
      481 gaccatgaaa ctggaaagcg caaaagttag ccgtgttttt tgcaccggta aaaaactggt
      541 gtatatcctg aaaacccgtc tgggtcgtac cattaaagca accgcaaatc atcgttttct
      601 gaccattgat ggttggaaac gtctggatga actgagcctg aaagaacata ttgcactgcc
      661 tcgtaaactg gaaagtagca gcctgcaact gagtccggaa attgaaaaac tgagccagag
      721 cgatatctat tgggatagca ttgttagcat taccgaaacc ggtgttgaag aagtttttga
      781 tctgaccgtt ccgggtccgc acaattttgt tgcaaatgat attattgtgc acaactgcac
      841 cctggaaaaa ctgatgaaag catttgaaag cctgaaaagc tttcagcagc agcaacaaca
      901 gcagcaacag caacagcagc agcagcagca acaacagcag cagcaacagc aacaacaaca
      961 acaacagcag caacaacagc aacagcagca acaacaacag caacagcaac aaccgcctcc
     1021 gcctccgcca cctcctcctc caccacagct gcctcagcct ccgcctcagt gccagccgct
     1081 gctgccacag cctcagccac ctccaccgcc tccacctcca cctccgggtc cggcagttgc
     1141 agaagaaccg ctgcatcgtc cgtaataact gcaggaaggg gatccggctg ctaacaaagc
     1201 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     1261 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggataact
     1321 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1381 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     1441 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     1501 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     1561 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     1621 gagagttttc gccccgaaga acgtttccca atgatgagca cttttaaagt tctgctatgt
     1681 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     1741 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     1801 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     1861 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     1921 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     1981 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     2041 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     2101 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     2161 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     2221 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     2281 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     2341 atactttaga ttgatttacc ccggttgata atcagaaaag ccccaaaaac aggaagattg
     2401 tataagcaaa tatttaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt
     2461 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     2521 aagaatagcc cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2581 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2641 gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     2701 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     2761 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     2821 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg taaaaggatc
     2881 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2941 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3001 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3061 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3121 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3181 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3241 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3301 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3361 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3421 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3481 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3541 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3601 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3661 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3721 cgcagcgagt cagtgagcga ggaagctatg gtgcactctc agtacaatct gctctgatgc
     3781 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     3841 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     3901 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     3961 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgcag cgattcacag
     4021 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
     4081 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
     4141 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
     4201 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
     4261 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagccgaac
     4321 gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg
     4381 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
     4441 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
     4501 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
     4561 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     4621 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
     4681 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4741 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
     4801 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
     4861 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
     4921 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac
     4981 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
     5041 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
     5101 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
     5161 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
     5221 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
     5281 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
     5341 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
     5401 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
     5461 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
     5521 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
     5581 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
     5641 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
     5701 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
     5761 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
     5821 aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct
     5881 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
     5941 gtgcatgccg gcatgccgcc ctttcgtctt caagaattaa ttcccaattc cccaggcatc
     6001 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     6061 tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac
     6121 ggcccggagg gtggcgggca ggacg
//
