LOCUS       sequence-362048-        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: I71L
FEATURES             Location/Qualifiers
     source          1..4633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(897..1835)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     rep_origin      1917..2505
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2406..2425
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2580..2827)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2793..2811
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(2893..3267)
                     /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"
     promoter        complement(3286..3333)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(3340..3753)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      complement(3768..4014)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(4094..4111)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4127..4156)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
ORIGIN
        1 tgcgtacaag tcctttctaa tagctgcagc acacttcatg atagaattgt aagtagtctc
       61 gtggattcca catgcctcca ttcccaaaaa tgatggctgg aacagggcct caggacatcg
      121 aaatctttcg ttgccgatgg taataacctg accgtctggc aattcgtaag acttctcaag
      181 tgaactacta gatgcagctg tagccatttc ctgttcaaaa tcaagggcaa cgtaacacaa
      241 tttttcctta atatctctga caatctcacg ctcagcagtt gtggtaaagg agtaccctcg
      301 ttctgtaaga attttcataa ggtaatcagt caggtcacgg ccagccaaat ctagacgcaa
      361 aatagcatga ggtaaagcat aaccttcgta tattggtaca gtatgagaga ccccatctcc
      421 tgagtctaaa actattcccg tggttctgcc tgaagcatac aaggacaata cagcttggat
      481 ggcgacatac atggctggag tgttgaaagt ctcgaacatt atttgggtca ttttttctct
      541 attggctttt ggattaagag gagcctctgt caataagact ggatgctctt caggtgccac
      601 tctcaattcg ttgtagaatg tgtgatgcca gattttctcc atgtcatccc agttagtaac
      661 tataccatgc tccaatgggt acttcaatgt caatattcct cttttagatt gagcttcgtc
      721 tcccacgtaa gagtcttttt gacccatgcc gaccattacg ccttgatgac ggggtcgacc
      781 tacaatagaa gggaaaacag ctctaggggc gtcgtcccca gcaaaacctg ctttacacat
      841 acctgaaccg ttatccacaa ccaaggcggc tacctcctca tcacacatga attcctcgtt
      901 tcgaataatt agttgttttt tgatcttctc aagttgtcgt taaaagtcgt taaaatcaaa
      961 agcttgtcaa ttggaaccag tcgcaattat gaaagtaagc taataatgat gataaaaaaa
     1021 aaggtttaag acagggcagc ttccttctgt ttatatattg ctgtcaagta ggggttagaa
     1081 cagttaaatt ttgatcatga acgttaggct atcagcagta ttcccaccag aatcttggaa
     1141 gcatacaatg tggagacaat gcataatcat ccaaaaagcg ggtgtttccc catttgcgtt
     1201 tcggcacagg tgcaccgggg ttcagaagcg atagagagac tgcgctaagc attaatgaga
     1261 ttatttttga gcattcgtca atcaatacca aacaagacaa acggtatgcc gacttttgga
     1321 agtttctttt tgaccaactg gccgttagca tttcaacgaa ccaaacttag ttcatcttgg
     1381 atgagatcac gcttttgtca tattaggttc caagacagcg tttaaactgt cagttttggg
     1441 ccatttgggg aacatgaaac tatttgaccc cacactcaga aagccctcat ctggagtgat
     1501 gttcgggtgt aatgcggagc ttgttgcatt cggaaataaa caaacatgaa cctcgccagg
     1561 ggggccagga tagacaggct aataaagtca tggtgttagt agcctaatag aaggaattgg
     1621 aatgagcgag ctccaatcaa gcccaataac tgggctggtt tttcgatggc aaaagtgggt
     1681 gttgaggaga agaggagtgg aggtcctgcg tttgcaacgg tctgctgcta gtgtatcccc
     1741 tcctgttgcg tttggcactt atgtgtgaga atggacctgt ggatgtcgga tggcaaaaag
     1801 gtttcattca acctttcgtc tttggatgtt agatctgatc tcatgaccaa aatcccttaa
     1861 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1921 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1981 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2041 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     2101 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2161 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2221 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2281 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2341 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2401 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2461 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2521 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttggt ctccagcttg
     2581 caaattaaag ccttcgagcg tcccaaaacc ttctcaagca aggttttcag tataatgtta
     2641 catgcgtaca cgcgtctgta cagaaaaaaa agaaaaattt gaaatataaa taacgttctt
     2701 aatactaaca taactataaa aaaataaata gggacctaga cttcaggttg tctaactcct
     2761 tccttttcgg ttagagcgga tgtgggggga gggcgtgaat gtaagcgtga cataactaat
     2821 tacatgatat cgacaaagga aaagggggac ggatctccga ggcctcggac ccgtcgggcc
     2881 gcgtcggacg tgtcagtcct gctcctcggc cacgaagtgc acgcagttgc cggccgggtc
     2941 gcgcagggcg aactcccgcc cccacggctg ctcgccgatc tcggtcatgg ccggcccgga
     3001 ggcgtcccgg aagttcgtgg acacgacctc cgaccactcg gcgtacagct cgtccaggcc
     3061 gcgcacccac acccaggcca gggtgttgtc cggcaccacc tggtcctgga ccgcgctgat
     3121 gaacagggtc acgtcgtccc ggaccacacc ggcgaagtcg tcctccacga agtcccggga
     3181 gaacccgagc cggtcggtcc agaactcgac cgctccggcg acgtcgcgcg cggtgagcac
     3241 cggaacggca ctggtcaact tggccatggt ttagttcctc accttgtcgt attatactat
     3301 gccgatatac tatgccgatg attaattgtc aacaccgccc ttagattaga ttgctatgct
     3361 ttctttctaa tgagcaagaa gtaaaaaaag ttgtaataga acaagaaaaa tgaaactgaa
     3421 acttgagaaa ttgaagaccg tttattaact taaatatcaa tgggaggtca tcgaaagaga
     3481 aaaaaatcaa aaaaaaaaat tttcaagaaa aagaaacgtg ataaaaattt ttattgcctt
     3541 tttcgacgaa gaaaaagaaa cgaggcggtc tcttttttct tttccaaacc tttagtacgg
     3601 gtaattaacg acaccctaga ggaagaaaga ggggaaattt agtatgctgt gcttgggtgt
     3661 tttgaagtgg tacggcgatg cgcggagtcc gagaaaatct ggaagagtaa aaaaggagta
     3721 gaaacatttt gaagctatgg tgtgtggggg atccgcacaa acgaaggtct cacttaatct
     3781 tctgtactct gaagaggagt gggaaatacc aagaaaaaca tcaaactcga atgattttcc
     3841 caaaccccta ccacaagata ttcatcagct gcgagatagg ctgatcagga gcaagctcgt
     3901 acgagaagaa acaaaatgac aaaaaaaatc ctatactata taggttacaa ataaaaaagt
     3961 atcaaaaatg aagcctgcat ctctcaggca aatggcattc tgacatcctc ttgattagaa
     4021 tctagcaaga ccggtcttct cgtaagtgcc caacttgaac tgaggaacag tcatgtctaa
     4081 ggctacaaac tcaatgatga tgatgatgat ggtcgacggc gctattcaga tcctcttctg
     4141 agatgagttt ttgttctaga aagctggcgg ccgcactttc accagcctgc ttttcctgtt
     4201 caatcgtttc cttagatggc aatggattct tctcttgggt ctcagttttt ttcaatttag
     4261 atttatcaaa cttctctatc tcagccatgt cgggcttatc agaggcagaa cctccacttg
     4321 atcctccaga tcctcctcta gaagcgccct gaaacaatac ttcaagggat ccaccactac
     4381 ctccacttcc accaaagcac ttacggtgga cgatacttgg tcctgattca tcgtattcct
     4441 gtttagagat ccacatttgt tggaatgtac taagagaggc caatatagaa ccgcctatcc
     4501 aaacagaata cttacgctca ggtggtgcga tgattttaat tttcatagta gatggagcca
     4561 aggccgtgat ttccttttgc attctatctg caatacctgg gtacatcgta gttcctccac
     4621 tcaggaccgt att
//
