LOCUS       sequence-222659-        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-P80C
FEATURES             Location/Qualifiers
     source          1..6124
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      13..99
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     234..253
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        234..252
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    253..277
                     /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             292..314
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             325..342
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1184..1202)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1198..1245
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1305..1409
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1410..1478,1479..2270)
                     /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(1628..1647)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2312..2825)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2507..2528)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2719..2738
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2936..3524
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3425..3444
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3678..3695
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3717..3736)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3877..3899
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3893..4084)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4641..5723)
                     /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     5685..5704
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5724..5801)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      5954..6040
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttccccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
       61 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
      121 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggaat
      181 tggggatcgg aattaattcc cggtttaaac cggggatctc gatcccgcga aattaatacg
      241 actcactata ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa
      301 ctttaagaag gagatataca tatgcatcat catcatcacc atagcggtgc aattagcggt
      361 gatagcctga ttagcctggc aagcaccggt aaacgtgtta gcattaaaga tctgctggat
      421 gagaaagatt ttgagatttg ggccattaac gagcagacca tgaaactgga aagcgcaaaa
      481 gttagccgtg ttttttgcac cggtaaaaaa ctggtgtata tcctgaaaac ccgtctgggt
      541 cgtaccatta aagcaaccgc aaatcatcgt tttctgacca ttgatggttg gaaacgtctg
      601 gatgaactga gcctgaaaga acatattgca ctgcctcgta aactggaaag tagcagcctg
      661 caactgagtc cggaaattga aaaactgagc cagagcgata tctattggga tagcattgtt
      721 agcattaccg aaaccggtgt tgaagaagtt tttgatctga ccgttccggg tccgcacaat
      781 tttgttgcaa atgatattat tgtgcacaac tgcaccctgg aaaaactgat gaaagcattt
      841 gaaagcctga aaagctttca gcagcagcaa caacagcagc aacagcaaca gcagcagcag
      901 cagcaacaac agcagcagca acagcaacaa caacaacaac agcagcaaca acagcaacag
      961 cagcagcaac cgcctccgcc tccgccacct cctcctccac cacagctgcc tcagcctccg
     1021 cctcaggcac agcctctgct gccacagcct cagccacctc caccgcctcc acctccacct
     1081 ccgggttgcg cagttgcaga agaaccgctg catcgtccgt aactgcagga aggggatccg
     1141 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     1201 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
     1261 atatccggat aactacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1321 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     1381 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     1441 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     1501 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     1561 gcggtaagat ccttgagagt tttcgccccg aagaacgttt cccaatgatg agcactttta
     1621 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     1681 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     1741 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     1801 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     1861 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     1921 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     1981 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     2041 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     2101 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     2161 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     2221 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     2281 aagtttactc atatatactt tagattgatt taccccggtt gataatcaga aaagccccaa
     2341 aaacaggaag attgtataag caaatattta aattgtaaac gttaatattt tgttaaaatt
     2401 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     2461 cccttataaa tcaaaagaat agcccgagat agggttgagt gttgttccag tttggaacaa
     2521 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     2581 cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa
     2641 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     2701 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     2761 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     2821 cgcgtaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     2881 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2941 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3001 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3061 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     3121 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3181 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3241 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3301 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3361 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3421 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3481 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3541 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3601 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3661 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc tatggtgcac tctcagtaca
     3721 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     3781 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3841 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3901 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     3961 gcagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     4021 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     4081 tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac
     4141 gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt
     4201 gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc
     4261 aatgccagcc gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa
     4321 tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg
     4381 cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc
     4441 ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa
     4501 agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga
     4561 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
     4621 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4681 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg
     4741 gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga
     4801 gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg
     4861 gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag
     4921 atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc
     4981 tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt
     5041 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
     5101 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
     5161 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
     5221 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
     5281 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
     5341 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
     5401 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
     5461 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
     5521 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
     5581 atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg
     5641 ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca
     5701 tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
     5761 catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc
     5821 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
     5881 cgccgcaagg aatggtgcat gccggcatgc cgccctttcg tcttcaagaa ttaattccca
     5941 attccccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     6001 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa
     6061 cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggaa
     6121 ttaa
//
