LOCUS       sequence-362348-        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: I274L
FEATURES             Location/Qualifiers
     source          1..4633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      457..1045
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     946..965
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1120..1367)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     1333..1351
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(1433..1807)
                     /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(1826..1873)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(1880..2293)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      complement(2308..2554)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(2634..2651)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2667..2696)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(join(4070..4633,1..375))
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
ORIGIN
        1 tatttgaccc cacactcaga aagccctcat ctggagtgat gttcgggtgt aatgcggagc
       61 ttgttgcatt cggaaataaa caaacatgaa cctcgccagg ggggccagga tagacaggct
      121 aataaagtca tggtgttagt agcctaatag aaggaattgg aatgagcgag ctccaatcaa
      181 gcccaataac tgggctggtt tttcgatggc aaaagtgggt gttgaggaga agaggagtgg
      241 aggtcctgcg tttgcaacgg tctgctgcta gtgtatcccc tcctgttgcg tttggcactt
      301 atgtgtgaga atggacctgt ggatgtcgga tggcaaaaag gtttcattca acctttcgtc
      361 tttggatgtt agatctgatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
      421 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
      481 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
      541 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
      601 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
      661 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
      721 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
      781 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
      841 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
      901 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
      961 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     1021 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     1081 cttttgctgg ccttttgctc acatgttggt ctccagcttg caaattaaag ccttcgagcg
     1141 tcccaaaacc ttctcaagca aggttttcag tataatgtta catgcgtaca cgcgtctgta
     1201 cagaaaaaaa agaaaaattt gaaatataaa taacgttctt aatactaaca taactataaa
     1261 aaaataaata gggacctaga cttcaggttg tctaactcct tccttttcgg ttagagcgga
     1321 tgtgggggga gggcgtgaat gtaagcgtga cataactaat tacatgatat cgacaaagga
     1381 aaagggggac ggatctccga ggcctcggac ccgtcgggcc gcgtcggacg tgtcagtcct
     1441 gctcctcggc cacgaagtgc acgcagttgc cggccgggtc gcgcagggcg aactcccgcc
     1501 cccacggctg ctcgccgatc tcggtcatgg ccggcccgga ggcgtcccgg aagttcgtgg
     1561 acacgacctc cgaccactcg gcgtacagct cgtccaggcc gcgcacccac acccaggcca
     1621 gggtgttgtc cggcaccacc tggtcctgga ccgcgctgat gaacagggtc acgtcgtccc
     1681 ggaccacacc ggcgaagtcg tcctccacga agtcccggga gaacccgagc cggtcggtcc
     1741 agaactcgac cgctccggcg acgtcgcgcg cggtgagcac cggaacggca ctggtcaact
     1801 tggccatggt ttagttcctc accttgtcgt attatactat gccgatatac tatgccgatg
     1861 attaattgtc aacaccgccc ttagattaga ttgctatgct ttctttctaa tgagcaagaa
     1921 gtaaaaaaag ttgtaataga acaagaaaaa tgaaactgaa acttgagaaa ttgaagaccg
     1981 tttattaact taaatatcaa tgggaggtca tcgaaagaga aaaaaatcaa aaaaaaaaat
     2041 tttcaagaaa aagaaacgtg ataaaaattt ttattgcctt tttcgacgaa gaaaaagaaa
     2101 cgaggcggtc tcttttttct tttccaaacc tttagtacgg gtaattaacg acaccctaga
     2161 ggaagaaaga ggggaaattt agtatgctgt gcttgggtgt tttgaagtgg tacggcgatg
     2221 cgcggagtcc gagaaaatct ggaagagtaa aaaaggagta gaaacatttt gaagctatgg
     2281 tgtgtggggg atccgcacaa acgaaggtct cacttaatct tctgtactct gaagaggagt
     2341 gggaaatacc aagaaaaaca tcaaactcga atgattttcc caaaccccta ccacaagata
     2401 ttcatcagct gcgagatagg ctgatcagga gcaagctcgt acgagaagaa acaaaatgac
     2461 aaaaaaaatc ctatactata taggttacaa ataaaaaagt atcaaaaatg aagcctgcat
     2521 ctctcaggca aatggcattc tgacatcctc ttgattagaa tctagcaaga ccggtcttct
     2581 cgtaagtgcc caacttgaac tgaggaacag tcatgtctaa ggctacaaac tcaatgatga
     2641 tgatgatgat ggtcgacggc gctattcaga tcctcttctg agatgagttt ttgttctaga
     2701 aagctggcgg ccgcactttc accagcctgc ttttcctgtt caatcgtttc cttagatggc
     2761 aatggattct tctcttgggt ctcagttttt ttcaatttag atttatcaaa cttctctatc
     2821 tcagccatgt cgggcttatc agaggcagaa cctccacttg atcctccaga tcctcctcta
     2881 gaagcgccct gaaacaatac ttcaagggat ccaccactac ctccacttcc accaaagcac
     2941 ttacggtgga cgatacttgg tcctgattca tcgtattcct gtttagagat ccacatttgt
     3001 tggaatgtac taagagaggc caatatagaa ccgcctatcc aaacagaata cttacgctca
     3061 ggtggtgcga tgattttaat tttcatagta gatggagcca aggccgtgat ttccttttgc
     3121 attctatctg caatacctgg gtacatcgta gttcctccac tcaggaccgt atttgcgtac
     3181 aagtcctttc taatagctgc agcacacttc atgatagaat tgtaagtagt ctcgtgcaat
     3241 ccacatgcct ccattcccaa aaatgatggc tggaacaggg cctcaggaca tcgaaatctt
     3301 tcgttgccga tggtaataac ctgaccgtct ggcaattcgt aagacttctc aagtgaacta
     3361 ctagatgcag ctgtagccat ttcctgttca aaatcaaggg caacgtaaca caatttttcc
     3421 ttaatatctc tgacaatctc acgctcagca gttgtggtaa aggagtaccc tcgttctgta
     3481 agaattttca taaggtaatc agtcaggtca cggccagcca aatctagacg caaaatagca
     3541 tgaggtaaag cataaccttc gtatattggt acagtatgag agaccccatc tcctgagtct
     3601 aaaactattc ccgtggttct gcctgaagca tacaaggaca atacagcttg gatggcgaca
     3661 tacatggctg gagtgttgaa agtctcgaac attatttggg tcattttttc tctattggct
     3721 tttggattaa gaggagcctc tgtcaataag actggatgct cttcaggtgc cactctcaat
     3781 tcgttgtaga atgtgtgatg ccagattttc tccatgtcat cccagttagt aactatacca
     3841 tgctcaattg ggtacttcaa tgtcaatatt cctcttttag attgagcttc gtctcccacg
     3901 taagagtctt tttgacccat gccgaccatt acgccttgat gacggggtcg acctacaata
     3961 gaagggaaaa cagctctagg ggcgtcgtcc ccagcaaaac ctgctttaca catacctgaa
     4021 ccgttatcca caaccaaggc ggctacctcc tcatcacaca tgaattcctc gtttcgaata
     4081 attagttgtt ttttgatctt ctcaagttgt cgttaaaagt cgttaaaatc aaaagcttgt
     4141 caattggaac cagtcgcaat tatgaaagta agctaataat gatgataaaa aaaaaggttt
     4201 aagacagggc agcttccttc tgtttatata ttgctgtcaa gtaggggtta gaacagttaa
     4261 attttgatca tgaacgttag gctatcagca gtattcccac cagaatcttg gaagcataca
     4321 atgtggagac aatgcataat catccaaaaa gcgggtgttt ccccatttgc gtttcggcac
     4381 aggtgcaccg gggttcagaa gcgatagaga gactgcgcta agcattaatg agattatttt
     4441 tgagcattcg tcaatcaata ccaaacaaga caaacggtat gccgactttt ggaagtttct
     4501 ttttgaccaa ctggccgtta gcatttcaac gaaccaaact tagttcatct tggatgagat
     4561 cacgcttttg tcatattagg ttccaagaca gcgtttaaac tgtcagtttt gggccatttg
     4621 gggaacatga aac
//
