LOCUS       sequence-362188-        4633 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 4633)
  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 4633)
  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: I248L
FEATURES             Location/Qualifiers
     source          1..4633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             547..576
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             592..609
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      689..935
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        950..1363
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        1370..1417
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1436..1810
                     /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      1876..2123
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1892..1910)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(2198..2786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2278..2297)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2868..3806
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
ORIGIN
        1 ggaatccacg agactactta caattctatc atgaagtgtg ctgcagctat tagaaaggac
       61 ttgtacgcaa atacggtcct gagtggagga actacgatgt acccaggtat tgcagataga
      121 atgcaaaagg aaatcacggc cttggctcca tctactatga aaattaaaat catcgcacca
      181 cctgagcgta agtattctgt ttggataggc ggttctatat tggcctctct tagtacattc
      241 caacaaatgt ggatctctaa acaggaatac gatgaatcag gaccaagtat cgtccaccgt
      301 aagtgctttg gtggaagtgg aggtagtggt ggatcccttg aagtattgtt tcagggcgct
      361 tctagaggag gatctggagg atcaagtgga ggttctgcct ctgataagcc cgacatggct
      421 gagatagaga agtttgataa atctaaattg aaaaaaactg agacccaaga gaagaatcca
      481 ttgccatcta aggaaacgat tgaacaggaa aagcaggctg gtgaaagtgc ggccgccagc
      541 tttctagaac aaaaactcat ctcagaagag gatctgaata gcgccgtcga ccatcatcat
      601 catcatcatt gagtttgtag ccttagacat gactgttcct cagttcaagt tgggcactta
      661 cgagaagacc ggtcttgcta gattctaatc aagaggatgt cagaatgcca tttgcctgag
      721 agatgcaggc ttcatttttg atactttttt atttgtaacc tatatagtat aggatttttt
      781 ttgtcatttt gtttcttctc gtacgagctt gctcctgatc agcctatctc gcagctgatg
      841 aatatcttgt ggtaggggtt tgggaaaatc attcgagttt gatgtttttc ttggtatttc
      901 ccactcctct tcagagtaca gaagattaag tgagaccttc gtttgtgcgg atcccccaca
      961 caccatagct tcaaaatgtt tctactcctt ttttactctt ccagattttc tcggactccg
     1021 cgcatcgccg taccacttca aaacacccaa gcacagcata ctaaatttcc cctctttctt
     1081 cctctagggt gtcgttaatt acccgtacta aaggtttgga aaagaaaaaa gagaccgcct
     1141 cgtttctttt tcttcgtcga aaaaggcaat aaaaattttt atcacgtttc tttttcttga
     1201 aaattttttt ttttgatttt tttctctttc gatgacctcc cattgatatt taagttaata
     1261 aacggtcttc aatttctcaa gtttcagttt catttttctt gttctattac aacttttttt
     1321 acttcttgct cattagaaag aaagcatagc aatctaatct aagggcggtg ttgacaatta
     1381 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc
     1441 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt
     1501 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt
     1561 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac
     1621 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt
     1681 gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg
     1741 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga
     1801 gcaggactga cacgtccgac gcggcccgac gggtccgagg cctcggagat ccgtccccct
     1861 tttcctttgt cgatatcatg taattagtta tgtcacgctt acattcacgc cctcccccca
     1921 catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt
     1981 tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc
     2041 tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg
     2101 gacgctcgaa ggctttaatt tgcaagctgg agaccaacat gtgagcaaaa ggccagcaaa
     2161 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2221 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2281 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2341 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2401 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2461 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2521 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     2581 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     2641 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     2701 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     2761 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2821 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagatcagat ctaacatcca
     2881 aagacgaaag gttgaatgaa acctttttgc catccgacat ccacaggtcc attctcacac
     2941 ataagtgcca aacgcaacag gaggggatac actagcagca gaccgttgca aacgcaggac
     3001 ctccactcct cttctcctca acacccactt ttgccatcga aaaaccagcc cagttattgg
     3061 gcttgattgg agctcgctca ttccaattcc ttctattagg ctactaacac catgacttta
     3121 ttagcctgtc tatcctggcc cccctggcga ggttcatgtt tgtttatttc cgaatgcaac
     3181 aagctccgca ttacacccga acatcactcc agatgagggc tttctgagtg tggggtcaaa
     3241 tagtttcatg ttccccaaat ggcccaaaac tgacagttta aacgctgtct tggaacctaa
     3301 tatgacaaaa gcgtgatctc atccaagatg aactaagttt ggttcgttga aatgctaacg
     3361 gccagttggt caaaaagaaa cttccaaaag tcggcatacc gtttgtcttg tttggtattg
     3421 attgacgaat gctcaaaaat aatctcatta atgcttagcg cagtctctct atcgcttctg
     3481 aaccccggtg cacctgtgcc gaaacgcaaa tggggaaaca cccgcttttt ggatgattat
     3541 gcattgtctc cacattgtat gcttccaaga ttctggtggg aatactgctg atagcctaac
     3601 gttcatgatc aaaatttaac tgttctaacc cctacttgac agcaatatat aaacagaagg
     3661 aagctgccct gtcttaaacc ttttttttta tcatcattat tagcttactt tcataattgc
     3721 gactggttcc aattgacaag cttttgattt taacgacttt taacgacaac ttgagaagat
     3781 caaaaaacaa ctaattattc gaaacgagga attcatgtgt gatgaggagg tagccgcctt
     3841 ggttgtggat aacggttcag gtatgtgtaa agcaggtttt gctggggacg acgcccctag
     3901 agctgttttc ccttctattg taggtcgacc ccgtcatcaa ggcgtaatgg tcggcatggg
     3961 tcaaaaagac tcttacgtgg gagacgaagc tcaatctaaa agaggaatat tgacattgaa
     4021 gtacccaatt gagcatggta tagttactaa ctgggatgac atggagaaaa tctggcatca
     4081 cacattctac aacgaattga gagtggcacc tgaagagcat ccagtcttat tgacagaggc
     4141 tcctcttaat ccaaaagcca atagagaaaa aatgacccaa ataatgttcg agactttcaa
     4201 cactccagcc atgtatgtcg ccatccaagc tgtattgtcc ttgtatgctt caggcagaac
     4261 cacgggaata gttttagact caggagatgg ggtctctcat actgtaccaa tatacgaagg
     4321 ttatgcttta cctcatgcta ttttgcgtct agatttggct ggccgtgacc tgactgatta
     4381 ccttatgaaa attcttacag aacgagggta ctcctttacc acaactgctg agcgtgagat
     4441 tgtcagagat attaaggaaa aattgtgtta cgttgccctt gattttgaac aggaaatggc
     4501 tacagctgca tctagtagtt cacttgagaa gtcttacgaa ttgccagacg gtcaggtttt
     4561 gaccatcggc aacgaaagat ttcgatgtcc tgaggccctg ttccagccat catttttggg
     4621 aatggaggca tgt
//
