LOCUS       sequence-222709-        6058 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 6058)
  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 6058)
  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)-15Q-P70C
FEATURES             Location/Qualifiers
     source          1..6058
                     /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(1220..1238)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1234..1281
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1341..1445
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1446..1514,1515..2306)
                     /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(1664..1683)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2348..2861)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2543..2564)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2755..2774
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2972..3560
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3461..3480
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3714..3731
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3753..3772)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3913..3935
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3929..4120)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4677..5759)
                     /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     5721..5740
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5760..5837)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      join(5990..6058,1..18)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
       61 gggtggcggg caggacgccc gccataaact gccaggaatt 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 atcatcacca tcattcaggt gcaattagcg gtgatagcct gattagcctg
      481 gcaagcaccg gtaaacgtgt tagcattaaa gatctgctgg atgagaaaga ttttgagatt
      541 tgggccatta acgagcagac catgaaactg gaaagcgcaa aagttagccg tgttttttgt
      601 accggcaaaa aactggtgta tatcctgaaa acccgtctgg gtcgtaccat taaagcaacc
      661 gcaaatcatc gttttctgac cattgatggt tggaaacgtc tggatgaact gagcctgaaa
      721 gaacatattg cactgcctcg taaattagaa agcagcagcc tgcaactgag tccggaaatt
      781 gaaaaactgt cacagagcga tatctattgg gatagcattg tgagcattac cgaaaccggt
      841 gttgaagaag tttttgatct gaccgttccg ggtccgcaca attttgttgc aaatgatatt
      901 atcgtgcaca attgcaccct ggaaaagctg atgaaagcat ttgaatcgct gaaatccttt
      961 cagcagcaac aacaacagca acagcaacaa cagcagcagc agcagcctcc tccgcctccg
     1021 cctccaccac ctcctccgca gttaccgcag cctccgccac aggcacagcc gttactgccg
     1081 cagccgcaac cgtgtcctcc tcctccgcca cccccaccag gtccggcagt tgcagaagaa
     1141 ccgctgcatc gtccgtaact gcaggaaggg gatccggctg ctaacaaagc ccgaaaggaa
     1201 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     1261 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggataact acgtcaggtg
     1321 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     1381 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     1441 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     1501 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     1561 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     1621 gccccgaaga acgtttccca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     1681 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1741 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     1801 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1861 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     1921 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     1981 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     2041 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     2101 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     2161 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     2221 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     2281 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     2341 ttgatttacc ccggttgata atcagaaaag ccccaaaaac aggaagattg tataagcaaa
     2401 tatttaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     2461 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagcc
     2521 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     2581 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     2641 acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     2701 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     2761 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     2821 caccacaccc gccgcgctta atgcgccgct acagggcgcg taaaaggatc taggtgaaga
     2881 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2941 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3001 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3061 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     3121 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3181 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3241 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3301 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3361 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3421 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3481 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3541 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3601 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3661 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3721 cagtgagcga ggaagctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3781 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     3841 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     3901 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     3961 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgcag cgattcacag atgtctgcct
     4021 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     4081 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
     4141 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
     4201 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
     4261 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagccgaac gccagcaaga
     4321 cgtagcccag cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt
     4381 tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa
     4441 gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga
     4501 gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga
     4561 cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     4621 tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac
     4681 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4741 cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg
     4801 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
     4861 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
     4921 gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg
     4981 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
     5041 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
     5101 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
     5161 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
     5221 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
     5281 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
     5341 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
     5401 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
     5461 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
     5521 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
     5581 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
     5641 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
     5701 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
     5761 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
     5821 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa
     5881 gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgccg
     5941 gcatgccgcc ctttcgtctt caagaattaa ttcccaattc cccaggcatc aaataaaacg
     6001 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgct
//
