LOCUS       sequence-362385-        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: I212L
FEATURES             Location/Qualifiers
     source          1..4633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(85..102)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(118..147)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(1521..2459)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     rep_origin      2541..3129
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3030..3049
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3204..3451)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     3417..3435
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(3517..3891)
                     /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(3910..3957)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(3964..4377)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      complement(join(4392..4633,1..5))
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
ORIGIN
        1 cttgattaga atctagcaag accggtcttc tcgtaagtgc ccaacttgaa ctgaggaaca
       61 gtcatgtcta aggctacaaa ctcaatgatg atgatgatga tggtcgacgg cgctattcag
      121 atcctcttct gagatgagtt tttgttctag aaagctggcg gccgcacttt caccagcctg
      181 cttttcctgt tcaatcgttt ccttagatgg caatggattc ttctcttggg tctcagtttt
      241 tttcaattta gatttatcaa acttctctat ctcagccatg tcgggcttat cagaggcaga
      301 acctccactt gatcctccag atcctcctct agaagcgccc tgaaacaata cttcaaggga
      361 tccaccacta cctccacttc caccaaagca cttacggtgg acgatacttg gtcctgattc
      421 atcgtattcc tgtttagaga tccacatttg ttggaatgta ctaagagagg ccaatataga
      481 accgcctatc caaacagaat acttacgctc aggtggtgcg atgattttaa ttttcatagt
      541 agatggagcc aaggccgtga tttccttttg cattctatct gcaatacctg ggtacatcgt
      601 agttcctcca ctcaggaccg tatttgcgta caagtccttt ctaatagctg cagcacactt
      661 catgatagaa ttgtaagtag tctcgtggat tccacatgcc tccattccca aaaatgatgg
      721 ctggaacagg gcctcaggac atcgaaatct ttcgttgccg atggtaataa cctgaccgtc
      781 tggcaattcg taagacttct caagtgaact actagatgca gctgtagcca tttcctgttc
      841 aaaatcaagg gcaacgtaac acaatttttc cttcaaatct ctgacaatct cacgctcagc
      901 agttgtggta aaggagtacc ctcgttctgt aagaattttc ataaggtaat cagtcaggtc
      961 acggccagcc aaatctagac gcaaaatagc atgaggtaaa gcataacctt cgtatattgg
     1021 tacagtatga gagaccccat ctcctgagtc taaaactatt cccgtggttc tgcctgaagc
     1081 atacaaggac aatacagctt ggatggcgac atacatggct ggagtgttga aagtctcgaa
     1141 cattatttgg gtcatttttt ctctattggc ttttggatta agaggagcct ctgtcaataa
     1201 gactggatgc tcttcaggtg ccactctcaa ttcgttgtag aatgtgtgat gccagatttt
     1261 ctccatgtca tcccagttag taactatacc atgctcaatt gggtacttca atgtcaatat
     1321 tcctctttta gattgagctt cgtctcccac gtaagagtct ttttgaccca tgccgaccat
     1381 tacgccttga tgacggggtc gacctacaat agaagggaaa acagctctag gggcgtcgtc
     1441 cccagcaaaa cctgctttac acatacctga accgttatcc acaaccaagg cggctacctc
     1501 ctcatcacac atgaattcct cgtttcgaat aattagttgt tttttgatct tctcaagttg
     1561 tcgttaaaag tcgttaaaat caaaagcttg tcaattggaa ccagtcgcaa ttatgaaagt
     1621 aagctaataa tgatgataaa aaaaaaggtt taagacaggg cagcttcctt ctgtttatat
     1681 attgctgtca agtaggggtt agaacagtta aattttgatc atgaacgtta ggctatcagc
     1741 agtattccca ccagaatctt ggaagcatac aatgtggaga caatgcataa tcatccaaaa
     1801 agcgggtgtt tccccatttg cgtttcggca caggtgcacc ggggttcaga agcgatagag
     1861 agactgcgct aagcattaat gagattattt ttgagcattc gtcaatcaat accaaacaag
     1921 acaaacggta tgccgacttt tggaagtttc tttttgacca actggccgtt agcatttcaa
     1981 cgaaccaaac ttagttcatc ttggatgaga tcacgctttt gtcatattag gttccaagac
     2041 agcgtttaaa ctgtcagttt tgggccattt ggggaacatg aaactatttg accccacact
     2101 cagaaagccc tcatctggag tgatgttcgg gtgtaatgcg gagcttgttg cattcggaaa
     2161 taaacaaaca tgaacctcgc caggggggcc aggatagaca ggctaataaa gtcatggtgt
     2221 tagtagccta atagaaggaa ttggaatgag cgagctccaa tcaagcccaa taactgggct
     2281 ggtttttcga tggcaaaagt gggtgttgag gagaagagga gtggaggtcc tgcgtttgca
     2341 acggtctgct gctagtgtat cccctcctgt tgcgtttggc acttatgtgt gagaatggac
     2401 ctgtggatgt cggatggcaa aaaggtttca ttcaaccttt cgtctttgga tgttagatct
     2461 gatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     2521 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2581 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2641 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     2701 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2761 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2821 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2881 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2941 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3001 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3061 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3121 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3181 gctcacatgt tggtctccag cttgcaaatt aaagccttcg agcgtcccaa aaccttctca
     3241 agcaaggttt tcagtataat gttacatgcg tacacgcgtc tgtacagaaa aaaaagaaaa
     3301 atttgaaata taaataacgt tcttaatact aacataacta taaaaaaata aatagggacc
     3361 tagacttcag gttgtctaac tccttccttt tcggttagag cggatgtggg gggagggcgt
     3421 gaatgtaagc gtgacataac taattacatg atatcgacaa aggaaaaggg ggacggatct
     3481 ccgaggcctc ggacccgtcg ggccgcgtcg gacgtgtcag tcctgctcct cggccacgaa
     3541 gtgcacgcag ttgccggccg ggtcgcgcag ggcgaactcc cgcccccacg gctgctcgcc
     3601 gatctcggtc atggccggcc cggaggcgtc ccggaagttc gtggacacga cctccgacca
     3661 ctcggcgtac agctcgtcca ggccgcgcac ccacacccag gccagggtgt tgtccggcac
     3721 cacctggtcc tggaccgcgc tgatgaacag ggtcacgtcg tcccggacca caccggcgaa
     3781 gtcgtcctcc acgaagtccc gggagaaccc gagccggtcg gtccagaact cgaccgctcc
     3841 ggcgacgtcg cgcgcggtga gcaccggaac ggcactggtc aacttggcca tggtttagtt
     3901 cctcaccttg tcgtattata ctatgccgat atactatgcc gatgattaat tgtcaacacc
     3961 gcccttagat tagattgcta tgctttcttt ctaatgagca agaagtaaaa aaagttgtaa
     4021 tagaacaaga aaaatgaaac tgaaacttga gaaattgaag accgtttatt aacttaaata
     4081 tcaatgggag gtcatcgaaa gagaaaaaaa tcaaaaaaaa aaattttcaa gaaaaagaaa
     4141 cgtgataaaa atttttattg cctttttcga cgaagaaaaa gaaacgaggc ggtctctttt
     4201 ttcttttcca aacctttagt acgggtaatt aacgacaccc tagaggaaga aagaggggaa
     4261 atttagtatg ctgtgcttgg gtgttttgaa gtggtacggc gatgcgcgga gtccgagaaa
     4321 atctggaaga gtaaaaaagg agtagaaaca ttttgaagct atggtgtgtg ggggatccgc
     4381 acaaacgaag gtctcactta atcttctgta ctctgaagag gagtgggaaa taccaagaaa
     4441 aacatcaaac tcgaatgatt ttcccaaacc cctaccacaa gatattcatc agctgcgaga
     4501 taggctgatc aggagcaagc tcgtacgaga agaaacaaaa tgacaaaaaa aatcctatac
     4561 tatataggtt acaaataaaa aagtatcaaa aatgaagcct gcatctctca ggcaaatggc
     4621 attctgacat cct
//
