LOCUS       sequence-362303-        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: I165L
FEATURES             Location/Qualifiers
     source          1..4633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(908..1846)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     rep_origin      1928..2516
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2417..2436
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2591..2838)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2804..2822
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(2904..3278)
                     /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(3297..3344)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(3351..3764)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      complement(3779..4025)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(4105..4122)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4138..4167)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
ORIGIN
        1 aggaccgtat ttgcgtacaa gtcctttcta atagctgcag cacacttcat gatagaattg
       61 taagtagtct cgtggattcc acatgcctcc attcccaaaa atgatggctg gaacagggcc
      121 tcaggacatc gaaatctttc gttgccgatg gtaataacct gaccgtctgg caattcgtaa
      181 gacttctcaa gtgaactact agatgcagct gtagccattt cctgttcaaa atcaagggca
      241 acgtaacaca atttttcctt aatatctctg acaatctcac gctcagcagt tgtggtaaag
      301 gagtaccctc gttctgtaag aattttcata aggtaatcag tcaggtcacg gccagccaaa
      361 tctagacgca aaatagcatg aggtaaagca taaccttcgt acaatggtac agtatgagag
      421 accccatctc ctgagtctaa aactattccc gtggttctgc ctgaagcata caaggacaat
      481 acagcttgga tggcgacata catggctgga gtgttgaaag tctcgaacat tatttgggtc
      541 attttttctc tattggcttt tggattaaga ggagcctctg tcaataagac tggatgctct
      601 tcaggtgcca ctctcaattc gttgtagaat gtgtgatgcc agattttctc catgtcatcc
      661 cagttagtaa ctataccatg ctcaattggg tacttcaatg tcaatattcc tcttttagat
      721 tgagcttcgt ctcccacgta agagtctttt tgacccatgc cgaccattac gccttgatga
      781 cggggtcgac ctacaataga agggaaaaca gctctagggg cgtcgtcccc agcaaaacct
      841 gctttacaca tacctgaacc gttatccaca accaaggcgg ctacctcctc atcacacatg
      901 aattcctcgt ttcgaataat tagttgtttt ttgatcttct caagttgtcg ttaaaagtcg
      961 ttaaaatcaa aagcttgtca attggaacca gtcgcaatta tgaaagtaag ctaataatga
     1021 tgataaaaaa aaaggtttaa gacagggcag cttccttctg tttatatatt gctgtcaagt
     1081 aggggttaga acagttaaat tttgatcatg aacgttaggc tatcagcagt attcccacca
     1141 gaatcttgga agcatacaat gtggagacaa tgcataatca tccaaaaagc gggtgtttcc
     1201 ccatttgcgt ttcggcacag gtgcaccggg gttcagaagc gatagagaga ctgcgctaag
     1261 cattaatgag attatttttg agcattcgtc aatcaatacc aaacaagaca aacggtatgc
     1321 cgacttttgg aagtttcttt ttgaccaact ggccgttagc atttcaacga accaaactta
     1381 gttcatcttg gatgagatca cgcttttgtc atattaggtt ccaagacagc gtttaaactg
     1441 tcagttttgg gccatttggg gaacatgaaa ctatttgacc ccacactcag aaagccctca
     1501 tctggagtga tgttcgggtg taatgcggag cttgttgcat tcggaaataa acaaacatga
     1561 acctcgccag gggggccagg atagacaggc taataaagtc atggtgttag tagcctaata
     1621 gaaggaattg gaatgagcga gctccaatca agcccaataa ctgggctggt ttttcgatgg
     1681 caaaagtggg tgttgaggag aagaggagtg gaggtcctgc gtttgcaacg gtctgctgct
     1741 agtgtatccc ctcctgttgc gtttggcact tatgtgtgag aatggacctg tggatgtcgg
     1801 atggcaaaaa ggtttcattc aacctttcgt ctttggatgt tagatctgat ctcatgacca
     1861 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     1921 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     1981 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     2041 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     2101 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     2161 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     2221 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     2281 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     2341 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     2401 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     2461 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     2521 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttgg
     2581 tctccagctt gcaaattaaa gccttcgagc gtcccaaaac cttctcaagc aaggttttca
     2641 gtataatgtt acatgcgtac acgcgtctgt acagaaaaaa aagaaaaatt tgaaatataa
     2701 ataacgttct taatactaac ataactataa aaaaataaat agggacctag acttcaggtt
     2761 gtctaactcc ttccttttcg gttagagcgg atgtgggggg agggcgtgaa tgtaagcgtg
     2821 acataactaa ttacatgata tcgacaaagg aaaaggggga cggatctccg aggcctcgga
     2881 cccgtcgggc cgcgtcggac gtgtcagtcc tgctcctcgg ccacgaagtg cacgcagttg
     2941 ccggccgggt cgcgcagggc gaactcccgc ccccacggct gctcgccgat ctcggtcatg
     3001 gccggcccgg aggcgtcccg gaagttcgtg gacacgacct ccgaccactc ggcgtacagc
     3061 tcgtccaggc cgcgcaccca cacccaggcc agggtgttgt ccggcaccac ctggtcctgg
     3121 accgcgctga tgaacagggt cacgtcgtcc cggaccacac cggcgaagtc gtcctccacg
     3181 aagtcccggg agaacccgag ccggtcggtc cagaactcga ccgctccggc gacgtcgcgc
     3241 gcggtgagca ccggaacggc actggtcaac ttggccatgg tttagttcct caccttgtcg
     3301 tattatacta tgccgatata ctatgccgat gattaattgt caacaccgcc cttagattag
     3361 attgctatgc tttctttcta atgagcaaga agtaaaaaaa gttgtaatag aacaagaaaa
     3421 atgaaactga aacttgagaa attgaagacc gtttattaac ttaaatatca atgggaggtc
     3481 atcgaaagag aaaaaaatca aaaaaaaaaa ttttcaagaa aaagaaacgt gataaaaatt
     3541 tttattgcct ttttcgacga agaaaaagaa acgaggcggt ctcttttttc ttttccaaac
     3601 ctttagtacg ggtaattaac gacaccctag aggaagaaag aggggaaatt tagtatgctg
     3661 tgcttgggtg ttttgaagtg gtacggcgat gcgcggagtc cgagaaaatc tggaagagta
     3721 aaaaaggagt agaaacattt tgaagctatg gtgtgtgggg gatccgcaca aacgaaggtc
     3781 tcacttaatc ttctgtactc tgaagaggag tgggaaatac caagaaaaac atcaaactcg
     3841 aatgattttc ccaaacccct accacaagat attcatcagc tgcgagatag gctgatcagg
     3901 agcaagctcg tacgagaaga aacaaaatga caaaaaaaat cctatactat ataggttaca
     3961 aataaaaaag tatcaaaaat gaagcctgca tctctcaggc aaatggcatt ctgacatcct
     4021 cttgattaga atctagcaag accggtcttc tcgtaagtgc ccaacttgaa ctgaggaaca
     4081 gtcatgtcta aggctacaaa ctcaatgatg atgatgatga tggtcgacgg cgctattcag
     4141 atcctcttct gagatgagtt tttgttctag aaagctggcg gccgcacttt caccagcctg
     4201 cttttcctgt tcaatcgttt ccttagatgg caatggattc ttctcttggg tctcagtttt
     4261 tttcaattta gatttatcaa acttctctat ctcagccatg tcgggcttat cagaggcaga
     4321 acctccactt gatcctccag atcctcctct agaagcgccc tgaaacaata cttcaaggga
     4381 tccaccacta cctccacttc caccaaagca cttacggtgg acgatacttg gtcctgattc
     4441 atcgtattcc tgtttagaga tccacatttg ttggaatgta ctaagagagg ccaatataga
     4501 accgcctatc caaacagaat acttacgctc aggtggtgcg atgattttaa ttttcatagt
     4561 agatggagcc aaggccgtga tttccttttg cattctatct gcaatacctg ggtacatcgt
     4621 agttcctcca ctc
//
