LOCUS       sequence-362192-        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: I208L
FEATURES             Location/Qualifiers
     source          1..4633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             629..658
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             674..691
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      771..1017
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1032..1445
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        1452..1499
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1518..1892
                     /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      1958..2205
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1974..1992)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(2280..2868)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2360..2379)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2950..3888
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
ORIGIN
        1 tcaggttatt accatcggca acgaaagatt tcgatgtcct gaggccctgt tccagccatc
       61 atttttggga atggaggcat gtggaatcca cgagactact tacaattcta tcatgaagtg
      121 tgctgcagct attagaaagg acttgtacgc aaatacggtc ctgagtggag gaactacgat
      181 gtacccaggt attgcagata gaatgcaaaa ggaaatcacg gccttggctc catctactat
      241 gaaaattaaa atcatcgcac cacctgagcg taagtattct gtttggatag gcggttctat
      301 attggcctct cttagtacat tccaacaaat gtggatctct aaacaggaat acgatgaatc
      361 aggaccaagt atcgtccacc gtaagtgctt tggtggaagt ggaggtagtg gtggatccct
      421 tgaagtattg tttcagggcg cttctagagg aggatctgga ggatcaagtg gaggttctgc
      481 ctctgataag cccgacatgg ctgagataga gaagtttgat aaatctaaat tgaaaaaaac
      541 tgagacccaa gagaagaatc cattgccatc taaggaaacg attgaacagg aaaagcaggc
      601 tggtgaaagt gcggccgcca gctttctaga acaaaaactc atctcagaag aggatctgaa
      661 tagcgccgtc gaccatcatc atcatcatca ttgagtttgt agccttagac atgactgttc
      721 ctcagttcaa gttgggcact tacgagaaga ccggtcttgc tagattctaa tcaagaggat
      781 gtcagaatgc catttgcctg agagatgcag gcttcatttt tgatactttt ttatttgtaa
      841 cctatatagt ataggatttt ttttgtcatt ttgtttcttc tcgtacgagc ttgctcctga
      901 tcagcctatc tcgcagctga tgaatatctt gtggtagggg tttgggaaaa tcattcgagt
      961 ttgatgtttt tcttggtatt tcccactcct cttcagagta cagaagatta agtgagacct
     1021 tcgtttgtgc ggatccccca cacaccatag cttcaaaatg tttctactcc ttttttactc
     1081 ttccagattt tctcggactc cgcgcatcgc cgtaccactt caaaacaccc aagcacagca
     1141 tactaaattt cccctctttc ttcctctagg gtgtcgttaa ttacccgtac taaaggtttg
     1201 gaaaagaaaa aagagaccgc ctcgtttctt tttcttcgtc gaaaaaggca ataaaaattt
     1261 ttatcacgtt tctttttctt gaaaattttt ttttttgatt tttttctctt tcgatgacct
     1321 cccattgata tttaagttaa taaacggtct tcaatttctc aagtttcagt ttcatttttc
     1381 ttgttctatt acaacttttt ttacttcttg ctcattagaa agaaagcata gcaatctaat
     1441 ctaagggcgg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     1501 aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg
     1561 acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg
     1621 aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg
     1681 accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt
     1741 acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga
     1801 ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact
     1861 gcgtgcactt cgtggccgag gagcaggact gacacgtccg acgcggcccg acgggtccga
     1921 ggcctcggag atccgtcccc cttttccttt gtcgatatca tgtaattagt tatgtcacgc
     1981 ttacattcac gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct
     2041 gaagtctagg tccctattta tttttttata gttatgttag tattaagaac gttatttata
     2101 tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa
     2161 accttgcttg agaaggtttt gggacgctcg aaggctttaa tttgcaagct ggagaccaac
     2221 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2281 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2341 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2401 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2461 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2521 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2581 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2641 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2701 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2761 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2821 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2881 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2941 atgagatcag atctaacatc caaagacgaa aggttgaatg aaaccttttt gccatccgac
     3001 atccacaggt ccattctcac acataagtgc caaacgcaac aggaggggat acactagcag
     3061 cagaccgttg caaacgcagg acctccactc ctcttctcct caacacccac ttttgccatc
     3121 gaaaaaccag cccagttatt gggcttgatt ggagctcgct cattccaatt ccttctatta
     3181 ggctactaac accatgactt tattagcctg tctatcctgg cccccctggc gaggttcatg
     3241 tttgtttatt tccgaatgca acaagctccg cattacaccc gaacatcact ccagatgagg
     3301 gctttctgag tgtggggtca aatagtttca tgttccccaa atggcccaaa actgacagtt
     3361 taaacgctgt cttggaacct aatatgacaa aagcgtgatc tcatccaaga tgaactaagt
     3421 ttggttcgtt gaaatgctaa cggccagttg gtcaaaaaga aacttccaaa agtcggcata
     3481 ccgtttgtct tgtttggtat tgattgacga atgctcaaaa ataatctcat taatgcttag
     3541 cgcagtctct ctatcgcttc tgaaccccgg tgcacctgtg ccgaaacgca aatggggaaa
     3601 cacccgcttt ttggatgatt atgcattgtc tccacattgt atgcttccaa gattctggtg
     3661 ggaatactgc tgatagccta acgttcatga tcaaaattta actgttctaa cccctacttg
     3721 acagcaatat ataaacagaa ggaagctgcc ctgtcttaaa cctttttttt tatcatcatt
     3781 attagcttac tttcataatt gcgactggtt ccaattgaca agcttttgat tttaacgact
     3841 tttaacgaca acttgagaag atcaaaaaac aactaattat tcgaaacgag gaattcatgt
     3901 gtgatgagga ggtagccgcc ttggttgtgg ataacggttc aggtatgtgt aaagcaggtt
     3961 ttgctgggga cgacgcccct agagctgttt tcccttctat tgtaggtcga ccccgtcatc
     4021 aaggcgtaat ggtcggcatg ggtcaaaaag actcttacgt gggagacgaa gctcaatcta
     4081 aaagaggaat attgacattg aagtacccaa ttgagcatgg tatagttact aactgggatg
     4141 acatggagaa aatctggcat cacacattct acaacgaatt gagagtggca cctgaagagc
     4201 atccagtctt attgacagag gctcctctta atccaaaagc caatagagaa aaaatgaccc
     4261 aaataatgtt cgagactttc aacactccag ccatgtatgt cgccatccaa gctgtattgt
     4321 ccttgtatgc ttcaggcaga accacgggaa tagttttaga ctcaggagat ggggtctctc
     4381 atactgtacc aatatacgaa ggttatgctt tacctcatgc tattttgcgt ctagatttgg
     4441 ctggccgtga cctgactgat taccttatga aaattcttac agaacgaggg tactccttta
     4501 ccacaactgc tgagcgtgag ttggtcagag atattaagga aaaattgtgt tacgttgccc
     4561 ttgattttga acaggaaatg gctacagctg catctagtag ttcacttgag aagtcttacg
     4621 aattgccaga cgg
//
