LOCUS       sequence-362194-        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: I34L
FEATURES             Location/Qualifiers
     source          1..4633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(107..124)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(140..169)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(1543..2481)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     rep_origin      2563..3151
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3052..3071
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3226..3473)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     3439..3457
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(3539..3913)
                     /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(3932..3979)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(3986..4399)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      complement(join(4414..4633,1..27))
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
ORIGIN
        1 gcaaatggca ttctgacatc ctcttgatta gaatctagca agaccggtct tctcgtaagt
       61 gcccaacttg aactgaggaa cagtcatgtc taaggctaca aactcaatga tgatgatgat
      121 gatggtcgac ggcgctattc agatcctctt ctgagatgag tttttgttct agaaagctgg
      181 cggccgcact ttcaccagcc tgcttttcct gttcaatcgt ttccttagat ggcaatggat
      241 tcttctcttg ggtctcagtt tttttcaatt tagatttatc aaacttctct atctcagcca
      301 tgtcgggctt atcagaggca gaacctccac ttgatcctcc agatcctcct ctagaagcgc
      361 cctgaaacaa tacttcaagg gatccaccac tacctccact tccaccaaag cacttacggt
      421 ggacgatact tggtcctgat tcatcgtatt cctgtttaga gatccacatt tgttggaatg
      481 tactaagaga ggccaatata gaaccgccta tccaaacaga atacttacgc tcaggtggtg
      541 cgatgatttt aattttcata gtagatggag ccaaggccgt gatttccttt tgcattctat
      601 ctgcaatacc tgggtacatc gtagttcctc cactcaggac cgtatttgcg tacaagtcct
      661 ttctaatagc tgcagcacac ttcatgatag aattgtaagt agtctcgtgg attccacatg
      721 cctccattcc caaaaatgat ggctggaaca gggcctcagg acatcgaaat ctttcgttgc
      781 cgatggtaat aacctgaccg tctggcaatt cgtaagactt ctcaagtgaa ctactagatg
      841 cagctgtagc catttcctgt tcaaaatcaa gggcaacgta acacaatttt tccttaatat
      901 ctctgacaat ctcacgctca gcagttgtgg taaaggagta ccctcgttct gtaagaattt
      961 tcataaggta atcagtcagg tcacggccag ccaaatctag acgcaaaata gcatgaggta
     1021 aagcataacc ttcgtatatt ggtacagtat gagagacccc atctcctgag tctaaaacta
     1081 ttcccgtggt tctgcctgaa gcatacaagg acaatacagc ttggatggcg acatacatgg
     1141 ctggagtgtt gaaagtctcg aacattattt gggtcatttt ttctctattg gcttttggat
     1201 taagaggagc ctctgtcaat aagactggat gctcttcagg tgccactctc aattcgttgt
     1261 agaatgtgtg atgccagatt ttctccatgt catcccagtt agtaactata ccatgctcaa
     1321 ttgggtactt caatgtcaat attcctcttt tagattgagc ttcgtctccc acgtaagagt
     1381 ctttttgacc catgccgacc attacgcctt gatgacgggg tcgacctacc aaagaaggga
     1441 aaacagctct aggggcgtcg tccccagcaa aacctgcttt acacatacct gaaccgttat
     1501 ccacaaccaa ggcggctacc tcctcatcac acatgaattc ctcgtttcga ataattagtt
     1561 gttttttgat cttctcaagt tgtcgttaaa agtcgttaaa atcaaaagct tgtcaattgg
     1621 aaccagtcgc aattatgaaa gtaagctaat aatgatgata aaaaaaaagg tttaagacag
     1681 ggcagcttcc ttctgtttat atattgctgt caagtagggg ttagaacagt taaattttga
     1741 tcatgaacgt taggctatca gcagtattcc caccagaatc ttggaagcat acaatgtgga
     1801 gacaatgcat aatcatccaa aaagcgggtg tttccccatt tgcgtttcgg cacaggtgca
     1861 ccggggttca gaagcgatag agagactgcg ctaagcatta atgagattat ttttgagcat
     1921 tcgtcaatca ataccaaaca agacaaacgg tatgccgact tttggaagtt tctttttgac
     1981 caactggccg ttagcatttc aacgaaccaa acttagttca tcttggatga gatcacgctt
     2041 ttgtcatatt aggttccaag acagcgttta aactgtcagt tttgggccat ttggggaaca
     2101 tgaaactatt tgaccccaca ctcagaaagc cctcatctgg agtgatgttc gggtgtaatg
     2161 cggagcttgt tgcattcgga aataaacaaa catgaacctc gccagggggg ccaggataga
     2221 caggctaata aagtcatggt gttagtagcc taatagaagg aattggaatg agcgagctcc
     2281 aatcaagccc aataactggg ctggtttttc gatggcaaaa gtgggtgttg aggagaagag
     2341 gagtggaggt cctgcgtttg caacggtctg ctgctagtgt atcccctcct gttgcgtttg
     2401 gcacttatgt gtgagaatgg acctgtggat gtcggatggc aaaaaggttt cattcaacct
     2461 ttcgtctttg gatgttagat ctgatctcat gaccaaaatc ccttaacgtg agttttcgtt
     2521 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2581 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2641 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2701 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2761 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2821 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2881 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2941 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3001 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3061 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3121 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3181 cctggccttt tgctggcctt ttgctcacat gttggtctcc agcttgcaaa ttaaagcctt
     3241 cgagcgtccc aaaaccttct caagcaaggt tttcagtata atgttacatg cgtacacgcg
     3301 tctgtacaga aaaaaaagaa aaatttgaaa tataaataac gttcttaata ctaacataac
     3361 tataaaaaaa taaataggga cctagacttc aggttgtcta actccttcct tttcggttag
     3421 agcggatgtg gggggagggc gtgaatgtaa gcgtgacata actaattaca tgatatcgac
     3481 aaaggaaaag ggggacggat ctccgaggcc tcggacccgt cgggccgcgt cggacgtgtc
     3541 agtcctgctc ctcggccacg aagtgcacgc agttgccggc cgggtcgcgc agggcgaact
     3601 cccgccccca cggctgctcg ccgatctcgg tcatggccgg cccggaggcg tcccggaagt
     3661 tcgtggacac gacctccgac cactcggcgt acagctcgtc caggccgcgc acccacaccc
     3721 aggccagggt gttgtccggc accacctggt cctggaccgc gctgatgaac agggtcacgt
     3781 cgtcccggac cacaccggcg aagtcgtcct ccacgaagtc ccgggagaac ccgagccggt
     3841 cggtccagaa ctcgaccgct ccggcgacgt cgcgcgcggt gagcaccgga acggcactgg
     3901 tcaacttggc catggtttag ttcctcacct tgtcgtatta tactatgccg atatactatg
     3961 ccgatgatta attgtcaaca ccgcccttag attagattgc tatgctttct ttctaatgag
     4021 caagaagtaa aaaaagttgt aatagaacaa gaaaaatgaa actgaaactt gagaaattga
     4081 agaccgttta ttaacttaaa tatcaatggg aggtcatcga aagagaaaaa aatcaaaaaa
     4141 aaaaattttc aagaaaaaga aacgtgataa aaatttttat tgcctttttc gacgaagaaa
     4201 aagaaacgag gcggtctctt ttttcttttc caaaccttta gtacgggtaa ttaacgacac
     4261 cctagaggaa gaaagagggg aaatttagta tgctgtgctt gggtgttttg aagtggtacg
     4321 gcgatgcgcg gagtccgaga aaatctggaa gagtaaaaaa ggagtagaaa cattttgaag
     4381 ctatggtgtg tgggggatcc gcacaaacga aggtctcact taatcttctg tactctgaag
     4441 aggagtggga aataccaaga aaaacatcaa actcgaatga ttttcccaaa cccctaccac
     4501 aagatattca tcagctgcga gataggctga tcaggagcaa gctcgtacga gaagaaacaa
     4561 aatgacaaaa aaaatcctat actatatagg ttacaaataa aaaagtatca aaaatgaagc
     4621 ctgcatctct cag
//
