LOCUS       sequence-222661-        6192 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 6192)
  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 6192)
  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-P90C
FEATURES             Location/Qualifiers
     source          1..6192
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      31..117
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     252..271
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        252..270
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    271..295
                     /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             310..332
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             343..360
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1270..1288)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1284..1331
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1391..1495
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1496..1564,1565..2356)
                     /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(1714..1733)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2398..2911)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2593..2614)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2805..2824
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      3022..3610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3511..3530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3764..3781
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3803..3822)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3963..3985
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3979..4170)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4727..5809)
                     /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     5771..5790
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5810..5887)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      6040..6126
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 taaactgcca ggaattaatt ccccaggcat caaataaaac gaaaggctca gtcgaaagac
       61 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
      121 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
      181 ccataaactg ccaggaattg gggatcggaa ttaattcccg gtttaaaccg gggatctcga
      241 tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct
      301 agaaataatt ttgtttaact ttaagaagga gatatacata tgcatcatca tcatcaccat
      361 agcggtgcaa ttagcggtga tagcctgatt agcctggcaa gcaccggtaa acgtgttagc
      421 attaaagatc tgctggatga gaaagatttt gagatttggg ccattaacga gcagaccatg
      481 aaactggaaa gcgcaaaagt tagccgtgtt ttttgcaccg gtaaaaaact ggtgtatatc
      541 ctgaaaaccc gtctgggtcg taccattaaa gcaaccgcaa atcatcgttt tctgaccatt
      601 gatggttgga aacgtctgga tgaactgagc ctgaaagaac atattgcact gcctcgtaaa
      661 ctggaaagta gcagcctgca actgagtccg gaaattgaaa aactgagcca gagcgatatc
      721 tattgggata gcattgttag cattaccgaa accggtgttg aagaagtttt tgatctgacc
      781 gttccgggtc cgcacaattt tgttgcaaat gatattattg tgcacaactg caccctggaa
      841 aaactgatga aagcatttga aagcctgaaa agctttcagc agcagcaaca acagcagcaa
      901 cagcaacagc agcagcagca gcaacaacag cagcagcaac agcaacaaca acaacaacag
      961 cagcaacaac agcaacagca gcagcaaccg cctccgcctc cgccacctcc tcctccacca
     1021 cagctgcctc agcctccgcc tcaggcacag cctctgctgc cacagcctca gccacctcca
     1081 ccgcctccac ctccacctcc gggtccggca gttgcagaag aaccgctgca tcgttgctaa
     1141 ctgcaggaag ggagaaccgc tgcatcgttg ctaactgcag gaagggagaa ccgctgcatc
     1201 gttgctaact gcaggaaggg gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     1261 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
     1321 ggggtttttt gctgaaagga ggaactatat ccggataact acgtcaggtg gcacttttcg
     1381 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1441 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1501 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1561 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     1621 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     1681 acgtttccca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     1741 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     1801 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     1861 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1921 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     1981 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     2041 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2101 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2161 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     2221 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2281 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2341 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttacc
     2401 ccggttgata atcagaaaag ccccaaaaac aggaagattg tataagcaaa tatttaaatt
     2461 gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2521 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagcc cgagataggg
     2581 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2641 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc acccaaatca
     2701 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2761 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2821 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     2881 gccgcgctta atgcgccgct acagggcgcg taaaaggatc taggtgaaga tcctttttga
     2941 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3001 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3061 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3121 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     3181 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3241 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3301 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3361 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3421 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3481 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3541 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3601 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3661 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3721 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3781 ggaagctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     3841 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     3901 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3961 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     4021 gcggtaaagc tcatcagcgt ggtcgtgcag cgattcacag atgtctgcct gttcatccgc
     4081 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     4141 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     4201 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     4261 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     4321 ggaccagaga aaaatcactc agggtcaatg ccagccgaac gccagcaaga cgtagcccag
     4381 cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
     4441 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
     4501 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
     4561 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
     4621 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
     4681 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     4741 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4801 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
     4861 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
     4921 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
     4981 cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa
     5041 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
     5101 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
     5161 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
     5221 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
     5281 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
     5341 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
     5401 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
     5461 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
     5521 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
     5581 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
     5641 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
     5701 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
     5761 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     5821 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
     5881 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt
     5941 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgccg gcatgccgcc
     6001 ctttcgtctt caagaattaa ttcccaattc cccaggcatc aaataaaacg aaaggctcag
     6061 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     6121 acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca
     6181 ggacgcccgc ca
//
