LOCUS       sequence-222814-        5977 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 5977)
  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 5977)
  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-A60C
FEATURES             Location/Qualifiers
     source          1..5977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      115..201
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     336..355
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        336..354
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    355..379
                     /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             394..416
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             427..444
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1322..1340)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1336..1383
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1443..1547
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1548..1616,1617..2408)
                     /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(1766..1785)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2450..2963)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2645..2666)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2857..2876
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      3074..3662
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3563..3582
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3816..3833
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3855..3874)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4015..4037
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4031..4222)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQVDICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4779..5861)
                     /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     5823..5842
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5862..5939)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
       61 tgccggcatg ccgccctttc gtcttcaaga attaattccc aattccccag gcatcaaata
      121 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
      181 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
      241 ggagggtggc gggcaggacg cccgccataa actgccagga attggggatc ggaattaatt
      301 cccggtttaa accggggatc tcgatcccgc gaaattaata cgactcacta taggggaatt
      361 gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata
      421 catatgcatc atcatcatca ccatagcggt gcaattagcg gtgatagcct gattagcctg
      481 gcaagcaccg gtaaacgtgt tagcattaaa gatctgctgg atgagaaaga ttttgagatt
      541 tgggccatta acgagcagac catgaaactg gaaagcgcaa aagttagccg tgttttttgc
      601 accggtaaaa aactggtgta tatcctgaaa acccgtctgg gtcgtaccat taaagcaacc
      661 gcaaatcatc gttttctgac cattgatggt tggaaacgtc tggatgaact gagcctgaaa
      721 gaacatattg cactgcctcg taaactggaa agtagcagcc tgcaactgag tccggaaatt
      781 gaaaaactga gccagagcga tatctattgg gatagcattg ttagcattac cgaaaccggt
      841 gttgaagaag tttttgatct gaccgttccg ggtccgcaca attttgttgc aaatgatatt
      901 attgtgcaca actgcaccct ggaaaaactg atgaaagcat ttgaaagcct gaaaagcttt
      961 cagcagcagc aacaacagca gcaacagcaa cagcagcagc agcagcaaca acagcagcag
     1021 caacagcaac aacaacaaca acagcagcaa caacagcaac agcagcaaca acaacagcaa
     1081 cagcagcagc agcaacaaca gcaacagcct ccgcctccgc caccgcctcc tccacctccg
     1141 cagctgccgc aacctccgcc tcagtgccag ccgctgctgc cgcagcctca acctcctcca
     1201 ccacctccac ctccaccgcc tggtccggca gttgcagaag aaccgctgca tcgtccgtaa
     1261 ctgcaggaag gggatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     1321 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1381 ttgctgaaag gaggaactat atccggataa ctacgtcagg tggcactttt cggggaaatg
     1441 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1501 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     1561 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1621 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     1681 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgtttcc
     1741 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     1801 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     1861 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     1921 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     1981 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     2041 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     2101 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     2161 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     2221 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     2281 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     2341 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     2401 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta ccccggttga
     2461 taatcagaaa agccccaaaa acaggaagat tgtataagca aatatttaaa ttgtaaacgt
     2521 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     2581 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag cccgagatag ggttgagtgt
     2641 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     2701 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat caagtttttt
     2761 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     2821 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     2881 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     2941 taatgcgccg ctacagggcg cgtaaaagga tctaggtgaa gatccttttt gataatctca
     3001 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3061 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3121 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3181 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     3241 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3301 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3361 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3421 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3481 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3541 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3601 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3661 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3721 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3781 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcta
     3841 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     3901 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     3961 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4021 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     4081 gctcatcagc gtggtcgtgc agcgattcac agatgtctac ctgttcatcc gcgtccagct
     4141 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     4201 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     4261 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     4321 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     4381 gaaaaatcac tcagggtcaa tgccagccga acgccagcaa gacgtagccc agcgcgtcgg
     4441 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     4501 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     4561 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
     4621 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     4681 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     4741 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4801 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4861 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     4921 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     4981 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     5041 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
     5101 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     5161 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     5221 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     5281 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     5341 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     5401 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     5461 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     5521 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     5581 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     5641 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     5701 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     5761 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     5821 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     5881 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc
     5941 gggatctcga cgctctccct tatgcgactc ctgcatt
//
