LOCUS       sequence-200847-        6037 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the human Huntingtin Exon1 protein containing 7Q in 
            E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6037)
  AUTHORS   Vieweg S, Ansaloni A, Wang ZM, Warner JB, Lashuel HA
  TITLE     An Intein-based Strategy for the Production of Tag-free Huntingtin 
            Exon 1 Proteins Enables New Insights into the Polyglutamine 
            Dependence of Httex1 Aggregation and Fibril Formation.
  JOURNAL   J Biol Chem. 2016 Jun 3;291(23):12074-86. doi: 
            10.1074/jbc.M116.713982. Epub 2016 Mar 21.
  PUBMED    27002149
REFERENCE   2  (bases 1 to 6037)
  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: 
            "10.1074/jbc.M116.713982"; journalName: "J Biol Chem"; date: 
            "2016-06-3- 3"; volume: "291"; issue: "23"; pages: "12074-86"
COMMENT     Sequence Label: pTWIN1-His6-Ssp-Httex1-7Q
FEATURES             Location/Qualifiers
     source          1..6037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      39..125
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     260..279
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        260..278
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    279..303
                     /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             318..340
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             351..368
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1123..1141)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1137..1184
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1244..1348
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1349..1417,1418..2209)
                     /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(1567..1586)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2251..2764)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2446..2467)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2658..2677
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2875..3463
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3364..3383
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3617..3634
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3656..3675)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3816..3838
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3832..4023)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4580..5662)
                     /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     5624..5643
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5663..5740)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      5893..5979
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcccgccata aactgccagg aattaattcc ccaggcatca aataaaacga aaggctcagt
       61 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
      121 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
      181 gacgcccgcc ataaactgcc aggaattggg gatcggaatt aattcccggt ttaaaccggg
      241 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
      301 tcccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg catcatcatc
      361 atcaccatag cggtgcaatt agcggtgata gcctgattag cctggcaagc accggtaaac
      421 gtgttagcat taaagatctg ctggatgaga aagattttga gatttgggcc attaacgagc
      481 agaccatgaa actggaaagc gcaaaagtta gccgtgtttt ttgcaccggt aaaaaactgg
      541 tgtatatcct gaaaacccgt ctgggtcgta ccattaaagc aaccgcaaat catcgttttc
      601 tgaccattga tggttggaaa cgtctggatg aactgagcct gaaagaacat attgcactgc
      661 ctcgtaaact ggaaagtagc agcctgcaac tgagtccgga aattgaaaaa ctgagccaga
      721 gcgatatcta ttgggatagc attgttagca ttaccgaaac cggtgttgaa gaagtttttg
      781 atctgaccgt tccgggtccg cacaattttg ttgcaaatga tattattgtg cacaacgcca
      841 ccctggaaaa actgatgaaa gcatttgaaa gcctgaaaag ctttcagcag cagcagcaac
      901 agcagcctcc tccgcctccg cctccaccac cccctccaca gctgcctcag cctccgccac
      961 aggcacagcc tctgctgcca cagccacaac cacccccacc accgccacca cctccgcctg
     1021 gtccggcagt tgcagaagaa ccgctgcatc gtccgtaata actgcaggaa ggggatccgg
     1081 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
     1141 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     1201 tatccggata actacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     1261 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     1321 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     1381 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     1441 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     1501 cggtaagatc cttgagagtt ttcgccccga agaacgtttc ccaatgatga gcacttttaa
     1561 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     1621 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1681 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     1741 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     1801 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     1861 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     1921 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1981 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     2041 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     2101 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     2161 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     2221 agtttactca tatatacttt agattgattt accccggttg ataatcagaa aagccccaaa
     2281 aacaggaaga ttgtataagc aaatatttaa attgtaaacg ttaatatttt gttaaaattc
     2341 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     2401 ccttataaat caaaagaata gcccgagata gggttgagtg ttgttccagt ttggaacaag
     2461 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     2521 gatggcccac tacgtgaacc atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa
     2581 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     2641 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     2701 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     2761 gcgtaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2821 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2881 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2941 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3001 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     3061 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3121 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3181 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3241 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3301 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3361 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3421 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3481 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3541 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3601 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagct atggtgcact ctcagtacaa
     3661 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     3721 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     3781 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     3841 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     3901 cagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     3961 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     4021 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
     4081 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
     4141 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
     4201 atgccagccg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc cggcgataat
     4261 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     4321 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     4381 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     4441 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     4501 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     4561 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4621 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     4681 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     4741 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     4801 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     4861 tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     4921 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     4981 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     5041 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     5101 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     5161 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     5221 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     5281 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     5341 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     5401 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     5461 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     5521 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     5581 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     5641 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     5701 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
     5761 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
     5821 gccgcaagga atggtgcatg ccggcatgcc gccctttcgt cttcaagaat taattcccaa
     5881 ttccccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     5941 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     6001 gttgcgaagc aacggcccgg agggtggcgg gcaggac
//
