LOCUS       sequence-362368-        5978 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To study dynamics of G-actin using NMR spectroscopy..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5978)
  AUTHORS   Ali R, Zahm JA, Rosen MK
  TITLE     Bound nucleotide can control the dynamic architecture of monomeric 
            actin.
  JOURNAL   Nat Struct Mol Biol. 2022 Apr;29(4):320-328. doi: 
            10.1038/s41594-022-00743-5. Epub 2022 Mar 24.
  PUBMED    35332323
REFERENCE   2  (bases 1 to 5978)
  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: 
            "10.1038/s41594-022-00743-5"; journalName: "Nat Struct Mol Biol"; 
            date: "2022-04"; volume: "29"; issue: "4"; pages: "320-328"
COMMENT     Sequence Label: I309L
FEATURES             Location/Qualifiers
     source          1..5978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             474..503
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             519..536
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      616..862
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        877..1290
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        1297..1344
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1363..1737
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      3064..3311
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3080..3098)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3386..3974)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3466..3485)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4056..4994
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             5012..5029
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5279..5299
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 aggaaatggc tacagctgca tctagtagtt cacttgagaa gtcttacgaa ttgccagacg
       61 gtcaggttat taccatcggc aacgaaagat ttcgatgtcc tgaggccctg ttccagccat
      121 catttttggg aatggaggca tgtggaatcc acgagactac ttacaattct atcatgaagt
      181 gtgctgcagc tattagaaag gacttgtacg caaatacggt cctgagtgga ggaactacga
      241 tgtacccagg tttggcagat agaatgcaaa aggaaatcac ggccttggct ccatctacta
      301 tgaaaattaa aatcatcgca ccacctgagc gtaagtattc tgtttggata ggcggttcta
      361 tattggcctc tcttagtaca ttccaacaaa tgtggatctc taaacaggaa tacgatgaat
      421 caggaccaag tatcgtccac cgtaagtgct tttaagcggc cgccagcttt ctagaacaaa
      481 aactcatctc agaagaggat ctgaatagcg ccgtcgacca tcatcatcat catcattgag
      541 tttgtagcct tagacatgac tgttcctcag ttcaagttgg gcacttacga gaagaccggt
      601 cttgctagat tctaatcaag aggatgtcag aatgccattt gcctgagaga tgcaggcttc
      661 atttttgata cttttttatt tgtaacctat atagtatagg attttttttg tcattttgtt
      721 tcttctcgta cgagcttgct cctgatcagc ctatctcgca gctgatgaat atcttgtggt
      781 aggggtttgg gaaaatcatt cgagtttgat gtttttcttg gtatttccca ctcctcttca
      841 gagtacagaa gattaagtga gaccttcgtt tgtgcggatc ccccacacac catagcttca
      901 aaatgtttct actccttttt tactcttcca gattttctcg gactccgcgc atcgccgtac
      961 cacttcaaaa cacccaagca cagcatacta aatttcccct ctttcttcct ctagggtgtc
     1021 gttaattacc cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt ttctttttct
     1081 tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt ttcttgaaaa tttttttttt
     1141 tgattttttt ctctttcgat gacctcccat tgatatttaa gttaataaac ggtcttcaat
     1201 ttctcaagtt tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgctcat
     1261 tagaaagaaa gcatagcaat ctaatctaag ggcggtgttg acaattaatc atcggcatag
     1321 tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt
     1381 gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg
     1441 ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg
     1501 accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg
     1561 tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc
     1621 cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc
     1681 gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac
     1741 gtccgacgcg gcccgacggg tccgatgatc ttacccagca atagtggaca cgcggctaag
     1801 tgagtaaact ctcagtcaga ggtgactcac atgacaaaaa cagtatcaac cagtaaaaaa
     1861 ccccgtaaac agcattcgcc tgaatttcgc agtgaagccc tgaagcttgc tgaacgcatc
     1921 ggtgttactg ccgcagcccg tgaactcagc ctgtatgaat cacagctcta caactggcgc
     1981 agtaaacagc aaaatcagca gacgtcttct gaacgtgaac tggagatgtc taccgagatt
     2041 gcacgtctca aacgccagct ggcagaacgg gatgaagagc tggctatcct ccaaaaggcc
     2101 gcgacatact tcgcgaagcg cctgaaatga agtatgtctt tattgaaaaa catcaggctg
     2161 agttcagcat caaagcaatg tgccgcgtgc tccgggtggc ccgcagcggc tggtatacgt
     2221 ggtgtcagcg gcggacaagg ataagcacgc gtcagcagtt ccgccaacac tgcgacagcg
     2281 ttgtcctcgc ggcttttacc cggtcaaaac agcgttacgg tgccccacgc ctgacggatg
     2341 aactgcgtgc tcagggttac ccctttaacg taaaaaccgt ggcggcaagc ctgcgccgtc
     2401 agggactgag ggcaaaggcc tcccggaagt tcagcccggt cagctaccgc gcacacggcc
     2461 tgcctgtgtc agaaaatctg ttggagcagg atttttacgc cagtggcccg aaccagaagt
     2521 gggcaggaga catcacgtac ttacgtacag atgaaggctg gctgtatctg gcagtggtca
     2581 ttgacctgtg gtcacgtgcc gttattggct ggtcaatgtc gccacgcatg acggcgcaac
     2641 tggcctgcga tgccctgcag atggcgctgt ggcggcgtaa gaggccccgg aacgttatcg
     2701 ttcacacgga tcgtggaggc cagtactgtt cagcagatta tcaggcgcaa ctgaagcggc
     2761 ataatctgcg tggaagtatg agcgcaaaag gttgctgcta cgataatgcc tgcgtggaaa
     2821 gcttctttca ttcgctgaaa gtggaatgta tccatggaga acactttatc agccgggaaa
     2881 taatgcgggc aacggtgttt aattatatcg aatgtgatta caatcggtgg cggcggcaca
     2941 gttggtgtgg cggcctcagt ccggaacaat ttgaaaacaa gaacctcgct taggcctgtg
     3001 tccatattac gtgggtagga tcacgaggcc tcggagatcc gtcccccttt tcctttgtcg
     3061 atatcatgta attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa
     3121 ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta
     3181 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg
     3241 tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg
     3301 ctttaatttg caagctggag accaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3361 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3421 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3481 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3541 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3601 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3661 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3721 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3781 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     3841 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3901 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3961 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4021 aaaactcacg ttaagggatt ttggtcatga gatcagatct aacatccaaa gacgaaaggt
     4081 tgaatgaaac ctttttgcca tccgacatcc acaggtccat tctcacacat aagtgccaaa
     4141 cgcaacagga ggggatacac tagcagcaga ccgttgcaaa cgcaggacct ccactcctct
     4201 tctcctcaac acccactttt gccatcgaaa aaccagccca gttattgggc ttgattggag
     4261 ctcgctcatt ccaattcctt ctattaggct actaacacca tgactttatt agcctgtcta
     4321 tcctggcccc cctggcgagg ttcatgtttg tttatttccg aatgcaacaa gctccgcatt
     4381 acacccgaac atcactccag atgagggctt tctgagtgtg gggtcaaata gtttcatgtt
     4441 ccccaaatgg cccaaaactg acagtttaaa cgctgtcttg gaacctaata tgacaaaagc
     4501 gtgatctcat ccaagatgaa ctaagtttgg ttcgttgaaa tgctaacggc cagttggtca
     4561 aaaagaaact tccaaaagtc ggcataccgt ttgtcttgtt tggtattgat tgacgaatgc
     4621 tcaaaaataa tctcattaat gcttagcgca gtctctctat cgcttctgaa ccccggtgca
     4681 cctgtgccga aacgcaaatg gggaaacacc cgctttttgg atgattatgc attgtctcca
     4741 cattgtatgc ttccaagatt ctggtgggaa tactgctgat agcctaacgt tcatgatcaa
     4801 aatttaactg ttctaacccc tacttgacag caatatataa acagaaggaa gctgccctgt
     4861 cttaaacctt tttttttatc atcattatta gcttactttc ataattgcga ctggttccaa
     4921 ttgacaagct tttgatttta acgactttta acgacaactt gagaagatca aaaaacaact
     4981 aattattcga aacgaggaat tcgccatggg tcaccatcat catcatcacg gcggctccgg
     5041 aggttctggt ggatctgcct ccgacaagcc cgacatggct gagattgaga agtttgacaa
     5101 atccaagtta aaaaaaacag agacccagga gaagaaccct ctacctagta aggagactat
     5161 tgaacaggaa aaacaagccg gtgaaagtgg tggatccggt ggctctggcg gttctggagg
     5221 aagtggtggt tcaggaggat caggtggatc tggcggatcc ggaggaagtg gtggatcaga
     5281 aaacttgtac tttcaaggcc atatgtgtga tgaggaggta gccgccttgg ttgtggataa
     5341 cggttcaggt atgtgtaaag caggttttgc tggggacgac gcccctagag ctgttttccc
     5401 ttctattgta ggtcgacccc gtcatcaagg cgtaatggtc ggcatgggtc aaaaagactc
     5461 ttacgtggga gacgaagctc aatctaaaag aggaatattg acattgaagt acccaattga
     5521 gcatggtata gttactaact gggatgacat ggagaaaatc tggcatcaca cattctacaa
     5581 cgaattgaga gtggcacctg aagagcatcc agtcttattg acagaggctc ctcttaatcc
     5641 aaaagccaat agagaaaaaa tgacccaaat aatgttcgag actttcaaca ctccagccat
     5701 gtatgtcgcc atccaagctg tattgtcctt gtatgcttca ggcagaacca cgggaatagt
     5761 tttagactca ggagatgggg tctctcatac tgtaccaata tacgaaggtt atgctttacc
     5821 tcatgctatt ttgcgtctag atttggctgg ccgtgacctg actgattacc ttatgaaaat
     5881 tcttacagaa cgagggtact cctttaccac aactgctgag cgtgagattg tcagagatat
     5941 taaggaaaaa ttgtgttacg ttgcccttga ttttgaac
//
