LOCUS       sequence-362156-        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: WT (DVD)
FEATURES             Location/Qualifiers
     source          1..5978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(897..917)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(1167..1184)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(1202..2140)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     rep_origin      2222..2810
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2711..2730
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2885..3132)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     3098..3116
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(4459..4833)
                     /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(4852..4899)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(4906..5319)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      complement(5334..5580)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(5660..5677)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(5693..5722)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
ORIGIN
        1 gtatttgcgt acaagtcctt tctaatagct gcagcacact tcatgataga attgtaagta
       61 gtctcgtgga ttccacatgc ctccattccc aaaaatgatg gctggaacag ggcctcagga
      121 catcgaaatc tttcgttgcc gatggtaata acctgaccgt ctggcaattc gtaagacttc
      181 tcaagtgaac tactagatgc agctgtagcc atttcctgtt caaaatcaag ggcaacgtaa
      241 cacaattttt ccttaatatc tctgacaatc tcacgctcag cagttgtggt aaaggagtac
      301 cctcgttctg taagaatttt cataaggtaa tcagtcaggt cacggccagc caaatctaga
      361 cgcaaaatag catgaggtaa agcataacct tcgtatattg gtacagtatg agagacccca
      421 tctcctgagt ctaaaactat tcccgtggtt ctgcctgaag catacaagga caatacagct
      481 tggatggcga catacatggc tggagtgttg aaagtctcga acattatttg ggtcattttt
      541 tctctattgg cttttggatt aagaggagcc tctgtcaata agactggatg ctcttcaggt
      601 gccactctca attcgttgta gaatgtgtga tgccagattt tctccatgtc atcccagtta
      661 gtaactatac catgctcaat tgggtacttc aatgtcaata ttcctctttt agattgagct
      721 tcgtctccca cgtaagagtc tttttgaccc atgccgacca ttacgccttg atgacggggt
      781 cgacctacaa tagaagggaa aacagctcta ggggcgtcgt ccccagcaaa acctgcttta
      841 cacatacctg aaccgttatc cacaaccaag gcggctacct cctcatcaca catatggcct
      901 tgaaagtaca agttttctga tccaccactt cctccggatc cgccagatcc acctgatcct
      961 cctgaaccac cacttcctcc agaaccgcca gagccaccgg atccaccact ttcaccggct
     1021 tgtttttcct gttcaatagt ctccttacta ggtagagggt tcttctcctg ggtctctgtt
     1081 ttttttaact tggatttgtc aaacttctca atctcagcca tgtcgggctt gtcggaggca
     1141 gatccaccag aacctccgga gccgccgtga tgatgatgat ggtgacccat ggcgaattcc
     1201 tcgtttcgaa taattagttg ttttttgatc ttctcaagtt gtcgttaaaa gtcgttaaaa
     1261 tcaaaagctt gtcaattgga accagtcgca attatgaaag taagctaata atgatgataa
     1321 aaaaaaaggt ttaagacagg gcagcttcct tctgtttata tattgctgtc aagtaggggt
     1381 tagaacagtt aaattttgat catgaacgtt aggctatcag cagtattccc accagaatct
     1441 tggaagcata caatgtggag acaatgcata atcatccaaa aagcgggtgt ttccccattt
     1501 gcgtttcggc acaggtgcac cggggttcag aagcgataga gagactgcgc taagcattaa
     1561 tgagattatt tttgagcatt cgtcaatcaa taccaaacaa gacaaacggt atgccgactt
     1621 ttggaagttt ctttttgacc aactggccgt tagcatttca acgaaccaaa cttagttcat
     1681 cttggatgag atcacgcttt tgtcatatta ggttccaaga cagcgtttaa actgtcagtt
     1741 ttgggccatt tggggaacat gaaactattt gaccccacac tcagaaagcc ctcatctgga
     1801 gtgatgttcg ggtgtaatgc ggagcttgtt gcattcggaa ataaacaaac atgaacctcg
     1861 ccaggggggc caggatagac aggctaataa agtcatggtg ttagtagcct aatagaagga
     1921 attggaatga gcgagctcca atcaagccca ataactgggc tggtttttcg atggcaaaag
     1981 tgggtgttga ggagaagagg agtggaggtc ctgcgtttgc aacggtctgc tgctagtgta
     2041 tcccctcctg ttgcgtttgg cacttatgtg tgagaatgga cctgtggatg tcggatggca
     2101 aaaaggtttc attcaacctt tcgtctttgg atgttagatc tgatctcatg accaaaatcc
     2161 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2221 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2281 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2341 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     2401 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2461 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2521 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2581 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2641 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2701 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2761 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2821 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttggtctcca
     2881 gcttgcaaat taaagccttc gagcgtccca aaaccttctc aagcaaggtt ttcagtataa
     2941 tgttacatgc gtacacgcgt ctgtacagaa aaaaaagaaa aatttgaaat ataaataacg
     3001 ttcttaatac taacataact ataaaaaaat aaatagggac ctagacttca ggttgtctaa
     3061 ctccttcctt ttcggttaga gcggatgtgg ggggagggcg tgaatgtaag cgtgacataa
     3121 ctaattacat gatatcgaca aaggaaaagg gggacggatc tccgaggcct cgtgatccta
     3181 cccacgtaat atggacacag gcctaagcga ggttcttgtt ttcaaattgt tccggactga
     3241 ggccgccaca ccaactgtgc cgccgccacc gattgtaatc acattcgata taattaaaca
     3301 ccgttgcccg cattatttcc cggctgataa agtgttctcc atggatacat tccactttca
     3361 gcgaatgaaa gaagctttcc acgcaggcat tatcgtagca gcaacctttt gcgctcatac
     3421 ttccacgcag attatgccgc ttcagttgcg cctgataatc tgctgaacag tactggcctc
     3481 cacggtccgt gtgaacgata acgttccggg gcctcttacg ccgccacagc gccatctgca
     3541 gggcatcgca ggccagttgc gccgtcatgc gtggcgacat tgaccagcca ataacggcac
     3601 gtgaccacag gtcaatgacc actgccagat acagccagcc ttcatctgta cgtaagtacg
     3661 tgatgtctcc tgcccacttc tggttcgggc cactggcgta aaaatcctgc tccaacagat
     3721 tttctgacac aggcaggccg tgtgcgcggt agctgaccgg gctgaacttc cgggaggcct
     3781 ttgccctcag tccctgacgg cgcaggcttg ccgccacggt ttttacgtta aaggggtaac
     3841 cctgagcacg cagttcatcc gtcaggcgtg gggcaccgta acgctgtttt gaccgggtaa
     3901 aagccgcgag gacaacgctg tcgcagtgtt ggcggaactg ctgacgcgtg cttatccttg
     3961 tccgccgctg acaccacgta taccagccgc tgcgggccac ccggagcacg cggcacattg
     4021 ctttgatgct gaactcagcc tgatgttttt caataaagac atacttcatt tcaggcgctt
     4081 cgcgaagtat gtcgcggcct tttggaggat agccagctct tcatcccgtt ctgccagctg
     4141 gcgtttgaga cgtgcaatct cggtagacat ctccagttca cgttcagaag acgtctgctg
     4201 attttgctgt ttactgcgcc agttgtagag ctgtgattca tacaggctga gttcacgggc
     4261 tgcggcagta acaccgatgc gttcagcaag cttcagggct tcactgcgaa attcaggcga
     4321 atgctgttta cggggttttt tactggttga tactgttttt gtcatgtgag tcacctctga
     4381 ctgagagttt actcacttag ccgcgtgtcc actattgctg ggtaagatca tcggacccgt
     4441 cgggccgcgt cggacgtgtc agtcctgctc ctcggccacg aagtgcacgc agttgccggc
     4501 cgggtcgcgc agggcgaact cccgccccca cggctgctcg ccgatctcgg tcatggccgg
     4561 cccggaggcg tcccggaagt tcgtggacac gacctccgac cactcggcgt acagctcgtc
     4621 caggccgcgc acccacaccc aggccagggt gttgtccggc accacctggt cctggaccgc
     4681 gctgatgaac agggtcacgt cgtcccggac cacaccggcg aagtcgtcct ccacgaagtc
     4741 ccgggagaac ccgagccggt cggtccagaa ctcgaccgct ccggcgacgt cgcgcgcggt
     4801 gagcaccgga acggcactgg tcaacttggc catggtttag ttcctcacct tgtcgtatta
     4861 tactatgccg atatactatg ccgatgatta attgtcaaca ccgcccttag attagattgc
     4921 tatgctttct ttctaatgag caagaagtaa aaaaagttgt aatagaacaa gaaaaatgaa
     4981 actgaaactt gagaaattga agaccgttta ttaacttaaa tatcaatggg aggtcatcga
     5041 aagagaaaaa aatcaaaaaa aaaaattttc aagaaaaaga aacgtgataa aaatttttat
     5101 tgcctttttc gacgaagaaa aagaaacgag gcggtctctt ttttcttttc caaaccttta
     5161 gtacgggtaa ttaacgacac cctagaggaa gaaagagggg aaatttagta tgctgtgctt
     5221 gggtgttttg aagtggtacg gcgatgcgcg gagtccgaga aaatctggaa gagtaaaaaa
     5281 ggagtagaaa cattttgaag ctatggtgtg tgggggatcc gcacaaacga aggtctcact
     5341 taatcttctg tactctgaag aggagtggga aataccaaga aaaacatcaa actcgaatga
     5401 ttttcccaaa cccctaccac aagatattca tcagctgcga gataggctga tcaggagcaa
     5461 gctcgtacga gaagaaacaa aatgacaaaa aaaatcctat actatatagg ttacaaataa
     5521 aaaagtatca aaaatgaagc ctgcatctct caggcaaatg gcattctgac atcctcttga
     5581 ttagaatcta gcaagaccgg tcttctcgta agtgcccaac ttgaactgag gaacagtcat
     5641 gtctaaggct acaaactcaa tgatgatgat gatgatggtc gacggcgcta ttcagatcct
     5701 cttctgagat gagtttttgt tctagaaagc tggcggccgc ttaaaagcac ttacggtgga
     5761 cgatacttgg tcctgattca tcgtattcct gtttagagat ccacatttgt tggaatgtac
     5821 taagagaggc caatatagaa ccgcctatcc aaacagaata cttacgctca ggtggtgcga
     5881 tgattttaat tttcatagta gatggagcca aggccgtgat ttccttttgc attctatctg
     5941 caatacctgg gtacatcgta gttcctccac tcaggacc
//
