LOCUS       sequence-222758-        5911 bp    DNA     circular SYN 15-APR-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 5911)
  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 5911)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-P70C
FEATURES             Location/Qualifiers
     source          1..5911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(734..752)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      748..795
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        855..959
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(960..1028,1029..1820)
                     /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(1178..1197)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(1862..2375)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2057..2078)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2269..2288
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2486..3074
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2975..2994
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3228..3245
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3267..3286)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3427..3449
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3443..3634)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4191..5273)
                     /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     5235..5254
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5274..5351)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      5504..5590
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5725..5744
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5725..5743
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5744..5768
                     /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             5783..5805
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5816..5833
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 gagaaagatt ttgagatttg ggccattaac gagcagacca tgaaactgga aagcgcaaaa
       61 gttagccgtg ttttttgcac cggtaaaaaa ctggtgtata tcctgaaaac ccgtctgggt
      121 cgtaccatta aagcaaccgc aaatcatcgt tttctgacca ttgatggttg gaaacgtctg
      181 gatgaactga gcctgaaaga acatattgca ctgcctcgta aactggaaag tagcagcctg
      241 caactgagtc cggaaattga aaaactgagc cagagcgata tctattggga tagcattgtt
      301 agcattaccg aaaccggtgt tgaagaagtt tttgatctga ccgttccggg tccgcacaat
      361 tttgttgcaa atgatattat cgtgcataat tgccagcagc agcaacaaca gcagcaacag
      421 caacagcagc agcagcagca acaacagcag cagcaacagc aacaacaaca acaacagcag
      481 caacaacagc aacagcagca acaacaacag caacagcagc ctccgcctcc gccacctcct
      541 cctccaccac ctcagctgcc tcagcctcct ccgcaggcac agcctctgct gccacagcct
      601 cagccatgcc ctccaccgcc accacctccg cctggtccag cagttgcaga agaaccgctg
      661 catcgtccgt aactgcagga aggggatccg gctgctaaca aagcccgaaa ggaagctgag
      721 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
      781 ttgaggggtt ttttgctgaa aggaggaact atatccggat aactacgtca ggtggcactt
      841 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
      901 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
      961 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     1021 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     1081 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     1141 aagaacgttt cccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     1201 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     1261 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     1321 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     1381 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     1441 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     1501 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     1561 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     1621 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     1681 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     1741 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     1801 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     1861 taccccggtt gataatcaga aaagccccaa aaacaggaag attgtataag caaatattta
     1921 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     1981 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agcccgagat
     2041 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2101 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccaa
     2161 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     2221 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2281 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     2341 acccgccgcg cttaatgcgc cgctacaggg cgcgtaaaag gatctaggtg aagatccttt
     2401 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2461 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2521 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2581 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     2641 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2701 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2761 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2821 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2881 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2941 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3001 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3061 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3121 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3181 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3241 gcgaggaagc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     3301 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
     3361 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     3421 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     3481 agctgcggta aagctcatca gcgtggtcgt gcagcgattc acagatgtct gcctgttcat
     3541 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     3601 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc
     3661 tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg
     3721 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc
     3781 ggcgggacca gagaaaaatc actcagggtc aatgccagcc gaacgccagc aagacgtagc
     3841 ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg
     3901 cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca
     3961 ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg
     4021 ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag
     4081 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     4141 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     4201 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4261 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     4321 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     4381 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
     4441 tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg
     4501 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     4561 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     4621 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     4681 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     4741 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
     4801 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     4861 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     4921 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     4981 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     5041 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     5101 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     5161 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     5221 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     5281 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     5341 tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc
     5401 cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gccggcatgc
     5461 cgccctttcg tcttcaagaa ttaattccca attccccagg catcaaataa aacgaaaggc
     5521 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     5581 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg
     5641 ggcaggacgc ccgccataaa ctgccaggaa ttggggatcg gaattaattc ccggtttaaa
     5701 ccggggatct cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata
     5761 acaattcccc tctagaaata attttgttta actttaagaa ggagatatac atatgcatca
     5821 tcatcatcac catagcggtg caattagcgg tgatagcctg attagcctgg caagcaccgg
     5881 taaacgtgtt agcattaaag atctgctgga t
//
