LOCUS       sequence-222655-        6124 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 6124)
  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 6124)
  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)-37Q-A60C
FEATURES             Location/Qualifiers
     source          1..6124
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     168..187
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        168..186
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    187..211
                     /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             226..248
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             259..276
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1118..1136)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1132..1179
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1239..1343
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1344..1412,1413..2204)
                     /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(1562..1581)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2246..2759)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2441..2462)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2653..2672
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2870..3458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3359..3378
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3612..3629
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3651..3670)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3811..3833
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3827..4018)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4575..5657)
                     /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     5619..5638
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5658..5735)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      5888..5974
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(6071..6124,1..33)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
       61 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gaattgggga
      121 tcggaattaa ttcccggttt aaaccgggga tctcgatccc gcgaaattaa tacgactcac
      181 tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt ttaactttaa
      241 gaaggagata tacatatgca tcatcatcat caccatagcg gtgcaattag cggtgatagc
      301 ctgattagcc tggcaagcac cggtaaacgt gttagcatta aagatctgct ggatgagaaa
      361 gattttgaga tttgggccat taacgagcag accatgaaac tggaaagcgc aaaagttagc
      421 cgtgtttttt gcaccggtaa aaaactggtg tatatcctga aaacccgtct gggtcgtacc
      481 attaaagcaa ccgcaaatca tcgttttctg accattgatg gttggaaacg tctggatgaa
      541 ctgagcctga aagaacatat tgcactgcct cgtaaactgg aaagtagcag cctgcaactg
      601 agtccggaaa ttgaaaaact gagccagagc gatatctatt gggatagcat tgttagcatt
      661 accgaaaccg gtgttgaaga agtttttgat ctgaccgttc cgggtccgca caattttgtt
      721 gcaaatgata ttattgtgca caactgcacc ctggaaaaac tgatgaaagc atttgaaagc
      781 ctgaaaagct ttcagcagca gcaacaacag cagcaacagc aacagcagca gcagcagcaa
      841 caacagcagc agcaacagca acaacaacaa caacagcagc aacaacagca acagcagcag
      901 caaccgcctc cgcctccgcc acctcctcct ccaccacagc tgcctcagcc tccgcctcag
      961 tgccagccgc tgctgccaca gcctcagcca cctccaccgc ctccacctcc acctccgggt
     1021 ccggcagttg cagaagaacc gctgcatcgt ccgtaactgc aggaagggga tccggctgct
     1081 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     1141 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     1201 ggataactac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1261 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1321 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     1381 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     1441 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     1501 agatccttga gagttttcgc cccgaagaac gtttcccaat gatgagcact tttaaagttc
     1561 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     1621 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     1681 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     1741 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     1801 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     1861 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     1921 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     1981 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     2041 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     2101 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     2161 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     2221 actcatatat actttagatt gatttacccc ggttgataat cagaaaagcc ccaaaaacag
     2281 gaagattgta taagcaaata tttaaattgt aaacgttaat attttgttaa aattcgcgtt
     2341 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     2401 taaatcaaaa gaatagcccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     2461 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     2521 cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     2581 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     2641 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     2701 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgta
     2761 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     2821 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2881 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2941 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3001 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3061 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3121 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3181 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3241 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3301 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3361 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3421 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3481 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3541 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3601 cgaccgagcg cagcgagtca gtgagcgagg aagctatggt gcactctcag tacaatctgc
     3661 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     3721 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3781 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3841 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgcagcg
     3901 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     3961 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     4021 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     4081 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     4141 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     4201 agccgaacgc cagcaagacg tagcccagcg cgtcggccgc catgccggcg ataatggcct
     4261 gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca
     4321 agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct
     4381 cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga
     4441 cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt
     4501 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     4561 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4621 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     4681 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     4741 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     4801 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc
     4861 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
     4921 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
     4981 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
     5041 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
     5101 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
     5161 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
     5221 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
     5281 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
     5341 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
     5401 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
     5461 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     5521 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     5581 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     5641 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     5701 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     5761 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
     5821 aaggaatggt gcatgccggc atgccgccct ttcgtcttca agaattaatt cccaattccc
     5881 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     5941 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc
     6001 gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca ggaattaatt
     6061 ccccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     6121 ttgt
//
