LOCUS       sequence-222750-        6082 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 6082)
  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 6082)
  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)-23Q-P80C
FEATURES             Location/Qualifiers
     source          1..6082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      44..130
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     265..284
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        265..283
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    284..308
                     /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             323..345
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             356..373
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1173..1191)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1187..1234
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1294..1398
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1399..1467,1468..2259)
                     /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(1617..1636)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2301..2814)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2496..2517)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2708..2727
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2925..3513
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3414..3433
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3667..3684
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3706..3725)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3866..3888
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3882..4073)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4630..5712)
                     /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     5674..5693
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5713..5790)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      5943..6029
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aggacgcccg ccataaactg ccaggaatta attccccagg catcaaataa aacgaaaggc
       61 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
      121 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg
      181 ggcaggacgc ccgccataaa ctgccaggaa ttggggatcg gaattaattc ccggtttaaa
      241 ccggggatct cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata
      301 acaattcccc tctagaaata attttgttta actttaagaa ggagatatac atatgcatca
      361 tcatcatcac catagcggtg caattagcgg tgatagcctg attagcctgg caagcaccgg
      421 taaacgtgtt agcattaaag atctgctgga tgagaaagat tttgagattt gggccattaa
      481 cgagcagacc atgaaactgg aaagcgcaaa agttagccgt gttttttgca ccggtaaaaa
      541 actggtgtat atcctgaaaa cccgtctggg tcgtaccatt aaagcaaccg caaatcatcg
      601 ttttctgacc attgatggtt ggaaacgtct ggatgaactg agcctgaaag aacatattgc
      661 actgcctcgt aaactggaaa gtagcagcct gcaactgagt ccggaaattg aaaaactgag
      721 ccagagcgat atctattggg atagcattgt tagcattacc gaaaccggtg ttgaagaagt
      781 ttttgatctg accgttccgg gtccgcacaa ttttgttgca aatgatatta ttgtgcacaa
      841 ctgcaccctg gaaaaactga tgaaagcatt tgaaagcctg aaaagctttc agcaacaaca
      901 gcagcaacag caacagcagc agcagcagca acaacagcag cagcaacagc aacaacaacc
      961 gcctccgcct ccgccacctc ctcctccacc acagctgcct cagcctccgc ctcaggcaca
     1021 gcctctgctg ccacagcctc agccacctcc accgcctcca cctccacctc cgggttgcgc
     1081 agttgcagaa gaaccgctgc atcgtccgta actgcaggaa ggggatccgg ctgctaacaa
     1141 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     1201 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata
     1261 actacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     1321 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1381 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1441 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1501 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1561 cttgagagtt ttcgccccga agaacgtttc ccaatgatga gcacttttaa agttctgcta
     1621 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     1681 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1741 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1801 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     1861 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     1921 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     1981 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2041 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2101 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2161 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2221 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2281 tatatacttt agattgattt accccggttg ataatcagaa aagccccaaa aacaggaaga
     2341 ttgtataagc aaatatttaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt
     2401 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     2461 caaaagaata gcccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     2521 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     2581 tacgtgaacc atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     2641 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     2701 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     2761 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtaaaagg
     2821 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     2881 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2941 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3001 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3061 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3121 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3181 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3241 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3301 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3361 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3421 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3481 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3541 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3601 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3661 gagcgcagcg agtcagtgag cgaggaagct atggtgcact ctcagtacaa tctgctctga
     3721 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     3781 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     3841 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     3901 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg cagcgattca
     3961 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     4021 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     4081 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     4141 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     4201 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagccg
     4261 aacgccagca agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc
     4321 tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt
     4381 ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg
     4441 aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc
     4501 ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag
     4561 gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt
     4621 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4681 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
     4741 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
     4801 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
     4861 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc
     4921 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
     4981 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
     5041 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
     5101 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
     5161 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
     5221 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
     5281 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
     5341 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
     5401 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
     5461 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
     5521 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
     5581 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     5641 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     5701 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
     5761 gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact
     5821 cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga
     5881 atggtgcatg ccggcatgcc gccctttcgt cttcaagaat taattcccaa ttccccaggc
     5941 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     6001 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc
     6061 aacggcccgg agggtggcgg gc
//
