LOCUS       sequence-225647-        6220 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Htt fragment containing amino acids 18-90, 
            with Q18C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6220)
  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 6220)
  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-Htt18-90(Q18C)-42Q-P90C
FEATURES             Location/Qualifiers
     source          1..6220
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     141..160
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        141..159
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    160..184
                     /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             199..221
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             232..249
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1187..1205)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1201..1248
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1308..1412
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1413..1481,1482..2273)
                     /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(1631..1650)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2315..2828)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2510..2531)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2722..2741
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2939..3527
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3428..3447
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3681..3698
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3720..3739)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3880..3902
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3896..4087)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4644..5726)
                     /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     5688..5707
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5727..5804)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      5957..6043
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(6140..6220,1..6)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca
       61 ggacgcccgc cataaactgc caggaattgg ggatcggaat taattcccgg tttaaaccgg
      121 ggatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
      181 ttcccctcta gaaataattt tgtttaactt taagaaggag atatacatat gcatcatcat
      241 catcaccata gcggtgcaat tagcggtgat agcctgatta gcctggcaag caccggtaaa
      301 cgtgttagca ttaaagatct gctggatgag aaagattttg agatttgggc cattaacgag
      361 cagaccatga aactggaaag cgcaaaagtt agccgtgttt tttgcaccgg taaaaaactg
      421 gtgtatatcc tgaaaacccg tctgggtcgt accattaaag caaccgcaaa tcatcgtttt
      481 ctgaccattg atggttggaa acgtctggat gaactgagcc tgaaagaaca tattgcactg
      541 cctcgtaaac tggaaagtag cagcctgcaa ctgagtccgg aaattgaaaa actgagccag
      601 agcgatatct attgggatag cattgttagc attaccgaaa ccggtgttga agaagttttt
      661 gatctgaccg ttccgggtcc gcacaatttt gttgcaaatg atattatcgt gcataattgc
      721 cagcagcagc aacaacagca gcaacagcaa cagcagcagc agcagcaaca acagcagcag
      781 caacagcaac aacaacaaca acagcagcaa caacagcaac agcagcaaca acaacagcaa
      841 cagcagcctc cgcctccgcc acctcctcct ccaccacctc agctgcctca gcctcctccg
      901 caggcacagc ctctgctgcc acagcctcag ccaccgcctc caccgccacc acctccgcct
      961 ggtccagcag ttgcagaaga accgctgcat cgttgctaac tgcaggaagg gggaaccgct
     1021 gcatcgttgc taactgcagg aagggaaccg ctgcatcgtt gctaactgca ggagaaccgc
     1081 tgcatcgttg ctaactgcag gaaagaaccg ctgcatcgtt gctaactgca ggaaggggat
     1141 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     1201 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     1261 actatatccg gataactacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1321 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1381 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1441 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1501 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1561 acagcggtaa gatccttgag agttttcgcc ccgaagaacg tttcccaatg atgagcactt
     1621 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     1681 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     1741 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     1801 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     1861 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     1921 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     1981 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2041 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2101 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2161 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2221 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2281 accaagttta ctcatatata ctttagattg atttaccccg gttgataatc agaaaagccc
     2341 caaaaacagg aagattgtat aagcaaatat ttaaattgta aacgttaata ttttgttaaa
     2401 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     2461 aatcccttat aaatcaaaag aatagcccga gatagggttg agtgttgttc cagtttggaa
     2521 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     2581 gggcgatggc ccactacgtg aaccatcacc caaatcaagt tttttggggt cgaggtgccg
     2641 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     2701 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     2761 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     2821 gggcgcgtaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     2881 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2941 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3001 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3061 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     3121 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3181 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3241 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3301 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3361 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3421 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3481 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3541 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3601 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3661 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agctatggtg cactctcagt
     3721 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     3781 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3841 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3901 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     3961 cgtgcagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     4021 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     4081 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     4141 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     4201 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
     4261 gtcaatgcca gccgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga
     4321 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     4381 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     4441 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     4501 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     4561 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     4621 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4681 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     4741 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     4801 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     4861 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     4921 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     4981 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     5041 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     5101 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     5161 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     5221 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     5281 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     5341 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     5401 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     5461 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     5521 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     5581 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     5641 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     5701 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     5761 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     5821 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     5881 cgccgccgca aggaatggtg catgccggca tgccgccctt tcgtcttcaa gaattaattc
     5941 ccaattcccc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     6001 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     6061 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     6121 gaattaattc cccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     6181 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
//
