LOCUS       sequence-334782-        4969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of yeast Ash1 truncation (encodes amino acids 
            420-500).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4969)
  AUTHORS   Martin EW, Holehouse AS, Grace CR, Hughes A, Pappu RV, Mittag T
  TITLE     Sequence Determinants of the Conformational Properties of an 
            Intrinsically Disordered Protein Prior to and upon Multisite 
            Phosphorylation.
  JOURNAL   J Am Chem Soc. 2016 Nov 30;138(47):15323-15335. doi: 
            10.1021/jacs.6b10272. Epub 2016 Nov 17.
  PUBMED    27807972
REFERENCE   2  (bases 1 to 4969)
  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.6b10272"; journalName: "J Am Chem Soc"; date: 
            "2016-11-30- 30"; volume: "138"; issue: "47"; pages: "15323-15335"
COMMENT     Sequence Label: Ash1
FEATURES             Location/Qualifiers
     source          1..4969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             93..110
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    120..144
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             147..167
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(418..442)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(495..513)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      509..556
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(922..927,928..944,945..950))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(977..995)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1063..1167
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1168..1236,1237..2028)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1386..1405)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2199..2787
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2688..2707
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2941..2958
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2973..3112
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3038..3057)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3198..3220
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3214..3405)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4867..4886
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atgtcgtact accatcacca tcaccatcac ctcgaatcaa
      121 caagtttgta caaaaaagca ggcttcgaaa acctgtattt tcagggcgcc tctgcatctt
      181 catctccatc tccctccaca ccgacgaaaa gtggcaagat gagatcaaga tcatcctcac
      241 ctgtgcgtcc caaggcatac actccaagcc caaggtcacc caattatcat agatttgctc
      301 tcgactctcc ccctcaaagc cctcgaagat catcaaattc aagcataacg aaaaagggaa
      361 gtagaaggtc cagtggctca tcacccactc gtcacaccac aagagtgtgc gtgtgagacc
      421 cagctttctt gtacaaagtg gttgattcga ggctgctaac aaagcccgaa aggaagctga
      481 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
      541 cttgaggggt tttttgctga aaggaggaac tatatccgga tatccacagg acgggtgtgg
      601 tcgccatgat cgcgtagtcg atagtggctc caagtagcga agcgagcagg actgggcggc
      661 ggccaaagcg gtcggacagt gctccgagaa cgggtgcgca tagaaattgc atcaacgcat
      721 atagcgctag cagcacgcca tagtgactgg cgatgctgtc ggaatggacg atatcccgca
      781 agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg gtgacggtgc
      841 cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg cgttagcaat
      901 ttaactgtga taaactaccg cattaaagct tatcgatgat aagctgtcaa acatgagaat
      961 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     1021 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     1081 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1141 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     1201 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     1261 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     1321 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     1381 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     1441 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     1501 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     1561 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     1621 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     1681 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     1741 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     1801 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     1861 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     1921 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     1981 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     2041 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     2101 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2161 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2221 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2281 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2341 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2401 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2461 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2521 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2581 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2641 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2701 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2761 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2821 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2881 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     2941 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     3001 ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat
     3061 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
     3121 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     3181 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     3241 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     3301 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     3361 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta
     3421 agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat
     3481 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
     3541 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa
     3601 tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat
     3661 aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac
     3721 cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc
     3781 gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct
     3841 caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc ccgagatgcg
     3901 ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag ggttggtttg
     3961 cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg tgaatccgtt
     4021 agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc accgcgacgc
     4081 aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa cccgttccat
     4141 gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccaatgat cgaagttagg
     4201 ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg
     4261 gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag aatcataatg
     4321 gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc
     4381 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     4441 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     4501 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     4561 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     4621 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgatc gacgctctcc
     4681 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
     4741 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     4801 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     4861 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     4921 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaat
//
