LOCUS       sequence-224744-        6160 bp    DNA     circular SYN 15-APR-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 6160)
  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 6160)
  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-Htt2-90(A2C)-49Q-P70C
FEATURES             Location/Qualifiers
     source          1..6160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      113..199
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     334..353
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        334..352
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    353..377
                     /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             392..414
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             425..442
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1320..1338)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1334..1381
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1441..1545
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1546..1614,1615..2406)
                     /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(1764..1783)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2448..2961)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2643..2664)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2855..2874
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      3072..3660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3561..3580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3814..3831
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3853..3872)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4013..4035
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4029..4220)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4777..5859)
                     /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     5821..5840
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5860..5937)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      join(6090..6160,1..16)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg
       61 gtggcgggca ggacgcccgc cataaactgc caggaattaa ttccccaggc atcaaataaa
      121 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
      181 tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg
      241 agggtggcgg gcaggacgcc cgccataaac tgccaggaat tggggatcgg aattaattcc
      301 cggtttaaac cggggatctc gatcccgcga aattaatacg actcactata ggggaattgt
      361 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatataca
      421 tatgcatcat catcaccatc attcaggtgc aattagcggt gatagcctga ttagcctggc
      481 aagcaccggt aaacgtgtta gcattaaaga tctgctggat gagaaagatt ttgagatttg
      541 ggccattaac gagcagacca tgaaactgga aagcgcaaaa gttagccgtg ttttttgtac
      601 cggcaaaaaa ctggtgtata tcctgaaaac ccgtctgggt cgtaccatta aagcaaccgc
      661 aaatcatcgt tttctgacca ttgatggttg gaaacgtctg gatgaactga gcctgaaaga
      721 acatattgca ctgcctcgta aattagaaag cagcagcctg caactgagtc cggaaattga
      781 aaaactgtca cagagcgata tctattggga tagcattgtg agcattaccg aaaccggtgt
      841 tgaagaagtt tttgatctga ccgttccggg tccgcacaat tttgttgcaa atgatattat
      901 cgtgcacaat tgcaccctgg aaaagctgat gaaagcattt gaatcgctga aatcctttca
      961 gcagcaacaa caacagcaac agcaacaaca gcagcagcaa cagcagcaac aacagcagca
     1021 gcaacaacaa caacagcagc aacagcaaca gcaacaacaa caacaacagc aacagcagca
     1081 acaacagcaa caacagcaac aacaaccgcc tccgcctccg ccacctcctc ctccaccgca
     1141 gttacctcag cctcctccgc aggcacagcc gctgctgcct cagccacagc cttgtccacc
     1201 gccaccccca ccacctccag gtccggcagt tgcagaagaa ccgctgcatc gtccgtaact
     1261 gcaggaaggg gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     1321 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     1381 gctgaaagga ggaactatat ccggataact acgtcaggtg gcacttttcg gggaaatgtg
     1441 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     1501 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     1561 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     1621 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     1681 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgtttccca
     1741 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     1801 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     1861 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     1921 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     1981 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     2041 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     2101 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     2161 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     2221 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     2281 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     2341 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     2401 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttacc ccggttgata
     2461 atcagaaaag ccccaaaaac aggaagattg tataagcaaa tatttaaatt gtaaacgtta
     2521 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     2581 ccgaaatcgg caaaatccct tataaatcaa aagaatagcc cgagataggg ttgagtgttg
     2641 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     2701 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc acccaaatca agttttttgg
     2761 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     2821 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     2881 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     2941 atgcgccgct acagggcgcg taaaaggatc taggtgaaga tcctttttga taatctcatg
     3001 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     3061 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3121 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3181 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     3241 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3301 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3361 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3421 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3481 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3541 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3601 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3661 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3721 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     3781 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagctatg
     3841 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     3901 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     3961 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     4021 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     4081 tcatcagcgt ggtcgtgcag cgattcacag atgtctgcct gttcatccgc gtccagctcg
     4141 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
     4201 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta
     4261 atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc
     4321 cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga
     4381 aaaatcactc agggtcaatg ccagccgaac gccagcaaga cgtagcccag cgcgtcggcc
     4441 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     4501 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     4561 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     4621 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     4681 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     4741 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4801 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4861 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     4921 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     4981 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     5041 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     5101 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     5161 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     5221 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     5281 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     5341 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     5401 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     5461 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     5521 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     5581 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     5641 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     5701 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     5761 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     5821 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     5881 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     5941 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     6001 ggccgttgag caccgccgcc gcaaggaatg gtgcatgccg gcatgccgcc ctttcgtctt
     6061 caagaattaa ttcccaattc cccaggcatc aaataaaacg aaaggctcag tcgaaagact
     6121 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
//
