LOCUS       sequence-222739-        6145 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 6145)
  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 6145)
  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)-43Q-P70C
FEATURES             Location/Qualifiers
     source          1..6145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     192..211
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        192..210
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    211..235
                     /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             250..272
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             283..300
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1163..1181)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1177..1224
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1284..1388
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1389..1457,1458..2249)
                     /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(1607..1626)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2291..2804)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2486..2507)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2698..2717
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      2915..3503
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3404..3423
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3657..3674
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3696..3715)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3856..3878
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3872..4063)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4620..5702)
                     /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     5664..5683
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5703..5780)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      5933..6019
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(6116..6145,1..57)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
       61 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
      121 ccataaactg ccaggaattg gggatcggaa ttaattcccg gtttaaaccg gggatctcga
      181 tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct
      241 agaaataatt ttgtttaact ttaagaagga gatatacata tgcatcatca tcatcaccat
      301 agcggtgcaa ttagcggtga tagcctgatt agcctggcaa gcaccggtaa acgtgttagc
      361 attaaagatc tgctggatga gaaagatttt gagatttggg ccattaacga gcagaccatg
      421 aaactggaaa gcgcaaaagt tagccgtgtt ttttgcaccg gtaaaaaact ggtgtatatc
      481 ctgaaaaccc gtctgggtcg taccattaaa gcaaccgcaa atcatcgttt tctgaccatt
      541 gatggttgga aacgtctgga tgaactgagc ctgaaagaac atattgcact gcctcgtaaa
      601 ctggaaagta gcagcctgca actgagtccg gaaattgaaa aactgagcca gagcgatatc
      661 tattgggata gcattgttag cattaccgaa accggtgttg aagaagtttt tgatctgacc
      721 gttccgggtc cgcacaattt tgttgcaaat gatattattg tgcacaactg caccctggaa
      781 aaactgatga aagcatttga aagcctgaaa agctttcagc agcagcaaca acagcagcaa
      841 cagcaacagc agcagcagca gcaacaacag cagcagcaac agcaacaaca acaacaacag
      901 cagcaacaac agcaacagca gcaacaacaa cagcaacagc aacaaccgcc tccgcctccg
      961 ccacctcctc ctccaccaca gctgcctcag cctccgcctc aggcacagcc tctgctgcca
     1021 cagcctcagc catgcccacc gcctccacct ccacctccgg gtccggcagt tgcagaagaa
     1081 ccgctgcatc gtccgtaata actgcaggaa ggggatccgg ctgctaacaa agcccgaaag
     1141 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     1201 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata actacgtcag
     1261 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1321 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     1381 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     1441 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     1501 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     1561 ttcgccccga agaacgtttc ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     1621 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     1681 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     1741 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     1801 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     1861 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     1921 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     1981 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     2041 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     2101 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     2161 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     2221 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     2281 agattgattt accccggttg ataatcagaa aagccccaaa aacaggaaga ttgtataagc
     2341 aaatatttaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     2401 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     2461 gcccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     2521 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     2581 atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     2641 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     2701 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     2761 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtaaaagg atctaggtga
     2821 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     2881 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2941 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3001 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3061 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3121 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3181 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3241 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3301 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3361 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3421 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3481 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3541 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     3601 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3661 agtcagtgag cgaggaagct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     3721 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
     3781 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     3841 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     3901 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg cagcgattca cagatgtctg
     3961 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     4021 taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag
     4081 ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac
     4141 gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg
     4201 tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagccg aacgccagca
     4261 agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac
     4321 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg
     4381 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc
     4441 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
     4501 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     4561 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     4621 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4681 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     4741 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     4801 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     4861 catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc
     4921 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     4981 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     5041 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     5101 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     5161 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     5221 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     5281 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     5341 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     5401 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     5461 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     5521 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     5581 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     5641 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     5701 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     5761 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     5821 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg
     5881 ccggcatgcc gccctttcgt cttcaagaat taattcccaa ttccccaggc atcaaataaa
     5941 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     6001 tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg
     6061 agggtggcgg gcaggacgcc cgccataaac tgccaggaat taattcccca ggcatcaaat
     6121 aaaacgaaag gctcagtcga aagac
//
