LOCUS       sequence-362351-        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: I151L
FEATURES             Location/Qualifiers
     source          1..4633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(906..1844)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     rep_origin      1926..2514
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2415..2434
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2589..2836)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2802..2820
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(2902..3276)
                     /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(3295..3342)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(3349..3762)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      complement(3777..4023)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(4103..4120)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4136..4165)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
ORIGIN
        1 gaccgtattt gcgtacaagt cctttctaat agctgcagca cacttcatga tagaattgta
       61 agtagtctcg tggattccac atgcctccat tcccaaaaat gatggctgga acagggcctc
      121 aggacatcga aatctttcgt tgccgatggt aataacctga ccgtctggca attcgtaaga
      181 cttctcaagt gaactactag atgcagctgt agccatttcc tgttcaaaat caagggcaac
      241 gtaacacaat ttttccttaa tatctctgac aatctcacgc tcagcagttg tggtaaagga
      301 gtaccctcgt tctgtaagaa ttttcataag gtaatcagtc aggtcacggc cagccaaatc
      361 tagacgcaaa atagcatgag gtaaagcata accttcgtat attggtacag tatgagagac
      421 cccatctcct gagtctaaaa ccaatcccgt ggttctgcct gaagcataca aggacaatac
      481 agcttggatg gcgacataca tggctggagt gttgaaagtc tcgaacatta tttgggtcat
      541 tttttctcta ttggcttttg gattaagagg agcctctgtc aataagactg gatgctcttc
      601 aggtgccact ctcaattcgt tgtagaatgt gtgatgccag attttctcca tgtcatccca
      661 gttagtaact ataccatgct caattgggta cttcaatgtc aatattcctc ttttagattg
      721 agcttcgtct cccacgtaag agtctttttg acccatgccg accattacgc cttgatgacg
      781 gggtcgacct acaatagaag ggaaaacagc tctaggggcg tcgtccccag caaaacctgc
      841 tttacacata cctgaaccgt tatccacaac caaggcggct acctcctcat cacacatgaa
      901 ttcctcgttt cgaataatta gttgtttttt gatcttctca agttgtcgtt aaaagtcgtt
      961 aaaatcaaaa gcttgtcaat tggaaccagt cgcaattatg aaagtaagct aataatgatg
     1021 ataaaaaaaa aggtttaaga cagggcagct tccttctgtt tatatattgc tgtcaagtag
     1081 gggttagaac agttaaattt tgatcatgaa cgttaggcta tcagcagtat tcccaccaga
     1141 atcttggaag catacaatgt ggagacaatg cataatcatc caaaaagcgg gtgtttcccc
     1201 atttgcgttt cggcacaggt gcaccggggt tcagaagcga tagagagact gcgctaagca
     1261 ttaatgagat tatttttgag cattcgtcaa tcaataccaa acaagacaaa cggtatgccg
     1321 acttttggaa gtttcttttt gaccaactgg ccgttagcat ttcaacgaac caaacttagt
     1381 tcatcttgga tgagatcacg cttttgtcat attaggttcc aagacagcgt ttaaactgtc
     1441 agttttgggc catttgggga acatgaaact atttgacccc acactcagaa agccctcatc
     1501 tggagtgatg ttcgggtgta atgcggagct tgttgcattc ggaaataaac aaacatgaac
     1561 ctcgccaggg gggccaggat agacaggcta ataaagtcat ggtgttagta gcctaataga
     1621 aggaattgga atgagcgagc tccaatcaag cccaataact gggctggttt ttcgatggca
     1681 aaagtgggtg ttgaggagaa gaggagtgga ggtcctgcgt ttgcaacggt ctgctgctag
     1741 tgtatcccct cctgttgcgt ttggcactta tgtgtgagaa tggacctgtg gatgtcggat
     1801 ggcaaaaagg tttcattcaa cctttcgtct ttggatgtta gatctgatct catgaccaaa
     1861 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1921 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1981 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2041 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2101 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2161 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2221 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2281 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2341 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2401 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2461 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2521 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttggtc
     2581 tccagcttgc aaattaaagc cttcgagcgt cccaaaacct tctcaagcaa ggttttcagt
     2641 ataatgttac atgcgtacac gcgtctgtac agaaaaaaaa gaaaaatttg aaatataaat
     2701 aacgttctta atactaacat aactataaaa aaataaatag ggacctagac ttcaggttgt
     2761 ctaactcctt ccttttcggt tagagcggat gtggggggag ggcgtgaatg taagcgtgac
     2821 ataactaatt acatgatatc gacaaaggaa aagggggacg gatctccgag gcctcggacc
     2881 cgtcgggccg cgtcggacgt gtcagtcctg ctcctcggcc acgaagtgca cgcagttgcc
     2941 ggccgggtcg cgcagggcga actcccgccc ccacggctgc tcgccgatct cggtcatggc
     3001 cggcccggag gcgtcccgga agttcgtgga cacgacctcc gaccactcgg cgtacagctc
     3061 gtccaggccg cgcacccaca cccaggccag ggtgttgtcc ggcaccacct ggtcctggac
     3121 cgcgctgatg aacagggtca cgtcgtcccg gaccacaccg gcgaagtcgt cctccacgaa
     3181 gtcccgggag aacccgagcc ggtcggtcca gaactcgacc gctccggcga cgtcgcgcgc
     3241 ggtgagcacc ggaacggcac tggtcaactt ggccatggtt tagttcctca ccttgtcgta
     3301 ttatactatg ccgatatact atgccgatga ttaattgtca acaccgccct tagattagat
     3361 tgctatgctt tctttctaat gagcaagaag taaaaaaagt tgtaatagaa caagaaaaat
     3421 gaaactgaaa cttgagaaat tgaagaccgt ttattaactt aaatatcaat gggaggtcat
     3481 cgaaagagaa aaaaatcaaa aaaaaaaatt ttcaagaaaa agaaacgtga taaaaatttt
     3541 tattgccttt ttcgacgaag aaaaagaaac gaggcggtct cttttttctt ttccaaacct
     3601 ttagtacggg taattaacga caccctagag gaagaaagag gggaaattta gtatgctgtg
     3661 cttgggtgtt ttgaagtggt acggcgatgc gcggagtccg agaaaatctg gaagagtaaa
     3721 aaaggagtag aaacattttg aagctatggt gtgtggggga tccgcacaaa cgaaggtctc
     3781 acttaatctt ctgtactctg aagaggagtg ggaaatacca agaaaaacat caaactcgaa
     3841 tgattttccc aaacccctac cacaagatat tcatcagctg cgagataggc tgatcaggag
     3901 caagctcgta cgagaagaaa caaaatgaca aaaaaaatcc tatactatat aggttacaaa
     3961 taaaaaagta tcaaaaatga agcctgcatc tctcaggcaa atggcattct gacatcctct
     4021 tgattagaat ctagcaagac cggtcttctc gtaagtgccc aacttgaact gaggaacagt
     4081 catgtctaag gctacaaact caatgatgat gatgatgatg gtcgacggcg ctattcagat
     4141 cctcttctga gatgagtttt tgttctagaa agctggcggc cgcactttca ccagcctgct
     4201 tttcctgttc aatcgtttcc ttagatggca atggattctt ctcttgggtc tcagtttttt
     4261 tcaatttaga tttatcaaac ttctctatct cagccatgtc gggcttatca gaggcagaac
     4321 ctccacttga tcctccagat cctcctctag aagcgccctg aaacaatact tcaagggatc
     4381 caccactacc tccacttcca ccaaagcact tacggtggac gatacttggt cctgattcat
     4441 cgtattcctg tttagagatc cacatttgtt ggaatgtact aagagaggcc aatatagaac
     4501 cgcctatcca aacagaatac ttacgctcag gtggtgcgat gattttaatt ttcatagtag
     4561 atggagccaa ggccgtgatt tccttttgca ttctatctgc aatacctggg tacatcgtag
     4621 ttcctccact cag
//
