LOCUS       sequence-222711-        6274 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 6274)
  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 6274)
  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-P90C
FEATURES             Location/Qualifiers
     source          1..6274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      42..128
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     263..282
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        263..281
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    282..306
                     /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             321..343
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             354..371
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1180..1198)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1194..1241
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1301..1405
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1406..1474,1475..2266)
                     /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(1624..1643)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2308..2821)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2503..2524)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2715..2734
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2932..3520
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3421..3440
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3674..3691
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3713..3732)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3873..3895
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3889..4080)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4637..5719)
                     /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     5681..5700
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5720..5797)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      5950..6036
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6133..6219
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gacgcccgcc ataaactgcc aggaattaat tccccaggca tcaaataaaa cgaaaggctc
       61 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
      121 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg
      181 caggacgccc gccataaact gccaggaatt ggggatcgga attaattccc ggtttaaacc
      241 ggggatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac
      301 aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacat atgcatcatc
      361 atcatcacca tagcggtgca attagcggtg atagcctgat tagcctggca agcaccggta
      421 aacgtgttag cattaaagat ctgctggatg agaaagattt tgagatttgg gccattaacg
      481 agcagaccat gaaactggaa agcgcaaaag ttagccgtgt tttttgcacc ggtaaaaaac
      541 tggtgtatat cctgaaaacc cgtctgggtc gtaccattaa agcaaccgca aatcatcgtt
      601 ttctgaccat tgatggttgg aaacgtctgg atgaactgag cctgaaagaa catattgcac
      661 tgcctcgtaa actggaaagt agcagcctgc aactgagtcc ggaaattgaa aaactgagcc
      721 agagcgatat ctattgggat agcattgtta gcattaccga aaccggtgtt gaagaagttt
      781 ttgatctgac cgttccgggt ccgcacaatt ttgttgcaaa tgatattatt gttcacaact
      841 gcaccctgga aaaactgatg aaagcatttg aaagcctgaa aagctttcag cagcagcaac
      901 aacagcagca acagcaacaa caacaacagc agcctccgcc tccgcctcct ccaccacctc
      961 ctccacagct gcctcagcct ccaccgcagg cacagcctct gctgccacag cctcagccac
     1021 ctccaccgcc accgcctcca cctccgggtc cagcagttgc agaagaaccg ctgcatcgtt
     1081 gctaactgca ggaaggggaa ccgctgcatc gttgctaact gcaggaaggg gatccggctg
     1141 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     1201 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     1261 ccggataact acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     1321 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1381 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     1441 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     1501 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     1561 taagatcctt gagagttttc gccccgaaga acgtttccca atgatgagca cttttaaagt
     1621 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     1681 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     1741 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     1801 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     1861 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     1921 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     1981 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2041 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2101 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     2161 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     2221 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     2281 ttactcatat atactttaga ttgatttacc ccggttgata atcagaaaag ccccaaaaac
     2341 aggaagattg tataagcaaa tatttaaatt gtaaacgtta atattttgtt aaaattcgcg
     2401 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     2461 tataaatcaa aagaatagcc cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     2521 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2581 ggcccactac gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca
     2641 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2701 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2761 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2821 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     2881 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2941 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3001 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3061 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3121 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3181 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3241 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3301 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3361 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3421 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3481 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3541 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     3601 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     3661 aacgaccgag cgcagcgagt cagtgagcga ggaagctatg gtgcactctc agtacaatct
     3721 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     3781 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3841 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3901 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgcag
     3961 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     4021 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     4081 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
     4141 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
     4201 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
     4261 ccagccgaac gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg cgataatggc
     4321 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
     4381 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
     4441 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
     4501 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     4561 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     4621 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4681 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     4741 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     4801 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     4861 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat
     4921 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     4981 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     5041 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     5101 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     5161 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     5221 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     5281 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     5341 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     5401 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     5461 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     5521 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     5581 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     5641 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     5701 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     5761 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
     5821 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
     5881 gcaaggaatg gtgcatgccg gcatgccgcc ctttcgtctt caagaattaa ttcccaattc
     5941 cccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     6001 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     6061 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggaattaa
     6121 ttccccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     6181 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     6241 gttgcgaagc aacggcccgg agggtggcgg gcag
//
