LOCUS Exported 6034 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Bacterial expression of Htt fragment containing amino acids 18-90, with Q18C. ACCESSION . VERSION . KEYWORDS pTWIN1-His6-Ssp-Htt18-90(Q18C)-22Q-A60C 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 May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6034 /organism="synthetic DNA construct" /mol_type="other DNA" 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 1229..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 5956..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 //