LOCUS       sequence-222795-        6034 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 6034)
  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 6034)
  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)-22Q-A60C
FEATURES             Location/Qualifiers
     source          1..6034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     143..162
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        143..161
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    162..186
                     /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             201..223
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             234..251
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1003..1021)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1017..1064
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1124..1228
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1229..1297,1298..2089)
                     /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(1447..1466)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2131..2644)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2326..2347)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2538..2557
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2755..3343
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3244..3263
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3497..3514
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3536..3555)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3696..3718
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3712..3903)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4460..5542)
                     /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     5504..5523
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5543..5620)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      5773..5859
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(5956..6034,1..8)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg
       61 caggacgccc gccataaact gccaggaatt ggggatcgga attaattccc ggtttaaacc
      121 ggggatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac
      181 aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacat atgcatcatc
      241 atcatcacca tagcggtgca attagcggtg atagcctgat tagcctggca agcaccggta
      301 aacgtgttag cattaaagat ctgctggatg agaaagattt tgagatttgg gccattaacg
      361 agcagaccat gaaactggaa agcgcaaaag ttagccgtgt tttttgcacc ggtaaaaaac
      421 tggtgtatat cctgaaaacc cgtctgggtc gtaccattaa agcaaccgca aatcatcgtt
      481 ttctgaccat tgatggttgg aaacgtctgg atgaactgag cctgaaagaa catattgcac
      541 tgcctcgtaa actggaaagt agcagcctgc aactgagtcc ggaaattgaa aaactgagcc
      601 agagcgatat ctattgggat agcattgtta gcattaccga aaccggtgtt gaagaagttt
      661 ttgatctgac cgttccgggt ccgcacaatt ttgttgcaaa tgatattatc gtgcataatt
      721 gccagcagca gcaacaacag cagcaacagc aacagcagca gcagcagcaa caacaacagc
      781 agcaacaacc gcctccgcct ccgccacctc ctcctccacc acagctgcct cagcctccgc
      841 ctcagtgtca gccgctgctg ccacagcctc agccacctcc accaccgcct ccacctccac
      901 ctggtccggc agttgcagaa gaaccgctgc atcgtccgta actgcaggaa ggggatccgg
      961 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
     1021 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     1081 tatccggata actacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     1141 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     1201 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     1261 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     1321 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     1381 cggtaagatc cttgagagtt ttcgccccga agaacgtttc ccaatgatga gcacttttaa
     1441 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     1501 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1561 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     1621 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     1681 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     1741 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     1801 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1861 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     1921 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     1981 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     2041 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     2101 agtttactca tatatacttt agattgattt accccggttg ataatcagaa aagccccaaa
     2161 aacaggaaga ttgtataagc aaatatttaa attgtaaacg ttaatatttt gttaaaattc
     2221 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     2281 ccttataaat caaaagaata gcccgagata gggttgagtg ttgttccagt ttggaacaag
     2341 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     2401 gatggcccac tacgtgaacc atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa
     2461 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     2521 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     2581 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     2641 gcgtaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2701 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2761 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2821 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2881 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     2941 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3001 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3061 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3121 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3181 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3241 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3301 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3361 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3421 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3481 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagct atggtgcact ctcagtacaa
     3541 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     3601 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     3661 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     3721 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     3781 cagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     3841 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     3901 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
     3961 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
     4021 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
     4081 atgccagccg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc cggcgataat
     4141 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     4201 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     4261 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     4321 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     4381 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     4441 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4501 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     4561 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     4621 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     4681 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     4741 tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     4801 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     4861 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     4921 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     4981 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     5041 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     5101 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     5161 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     5221 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     5281 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     5341 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     5401 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     5461 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     5521 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     5581 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
     5641 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
     5701 gccgcaagga atggtgcatg ccggcatgcc gccctttcgt cttcaagaat taattcccaa
     5761 ttccccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     5821 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     5881 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggaat
     5941 taattcccca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     6001 atctgttgtt tgtcggtgaa cgctctcctg agta
//
