LOCUS       sequence-362346-        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: I122L
FEATURES             Location/Qualifiers
     source          1..4633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             546..575
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             591..608
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      688..934
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        949..1362
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        1369..1416
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1435..1809
                     /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"
     terminator      1875..2122
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1891..1909)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(2197..2785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2277..2296)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2867..3805
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
ORIGIN
        1 gaatccacga gactacttac aattctatca tgaagtgtgc tgcagctatt agaaaggact
       61 tgtacgcaaa tacggtcctg agtggaggaa ctacgatgta cccaggtatt gcagatagaa
      121 tgcaaaagga aatcacggcc ttggctccat ctactatgaa aattaaaatc atcgcaccac
      181 ctgagcgtaa gtattctgtt tggataggcg gttctatatt ggcctctctt agtacattcc
      241 aacaaatgtg gatctctaaa caggaatacg atgaatcagg accaagtatc gtccaccgta
      301 agtgctttgg tggaagtgga ggtagtggtg gatcccttga agtattgttt cagggcgctt
      361 ctagaggagg atctggagga tcaagtggag gttctgcctc tgataagccc gacatggctg
      421 agatagagaa gtttgataaa tctaaattga aaaaaactga gacccaagag aagaatccat
      481 tgccatctaa ggaaacgatt gaacaggaaa agcaggctgg tgaaagtgcg gccgccagct
      541 ttctagaaca aaaactcatc tcagaagagg atctgaatag cgccgtcgac catcatcatc
      601 atcatcattg agtttgtagc cttagacatg actgttcctc agttcaagtt gggcacttac
      661 gagaagaccg gtcttgctag attctaatca agaggatgtc agaatgccat ttgcctgaga
      721 gatgcaggct tcatttttga tactttttta tttgtaacct atatagtata ggattttttt
      781 tgtcattttg tttcttctcg tacgagcttg ctcctgatca gcctatctcg cagctgatga
      841 atatcttgtg gtaggggttt gggaaaatca ttcgagtttg atgtttttct tggtatttcc
      901 cactcctctt cagagtacag aagattaagt gagaccttcg tttgtgcgga tcccccacac
      961 accatagctt caaaatgttt ctactccttt tttactcttc cagattttct cggactccgc
     1021 gcatcgccgt accacttcaa aacacccaag cacagcatac taaatttccc ctctttcttc
     1081 ctctagggtg tcgttaatta cccgtactaa aggtttggaa aagaaaaaag agaccgcctc
     1141 gtttcttttt cttcgtcgaa aaaggcaata aaaattttta tcacgtttct ttttcttgaa
     1201 aatttttttt tttgattttt ttctctttcg atgacctccc attgatattt aagttaataa
     1261 acggtcttca atttctcaag tttcagtttc atttttcttg ttctattaca acttttttta
     1321 cttcttgctc attagaaaga aagcatagca atctaatcta agggcggtgt tgacaattaa
     1381 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc
     1441 aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc
     1501 tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc
     1561 cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc
     1621 ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg
     1681 tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg
     1741 gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag
     1801 caggactgac acgtccgacg cggcccgacg ggtccgaggc ctcggagatc cgtccccctt
     1861 ttcctttgtc gatatcatgt aattagttat gtcacgctta cattcacgcc ctccccccac
     1921 atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt
     1981 ttttatagtt atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct
     2041 gtacagacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg
     2101 acgctcgaag gctttaattt gcaagctgga gaccaacatg tgagcaaaag gccagcaaaa
     2161 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2221 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2281 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2341 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2401 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2461 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2521 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2581 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2641 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2701 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2761 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2821 tcagtggaac gaaaactcac gttaagggat tttggtcatg agatcagatc taacatccaa
     2881 agacgaaagg ttgaatgaaa cctttttgcc atccgacatc cacaggtcca ttctcacaca
     2941 taagtgccaa acgcaacagg aggggataca ctagcagcag accgttgcaa acgcaggacc
     3001 tccactcctc ttctcctcaa cacccacttt tgccatcgaa aaaccagccc agttattggg
     3061 cttgattgga gctcgctcat tccaattcct tctattaggc tactaacacc atgactttat
     3121 tagcctgtct atcctggccc ccctggcgag gttcatgttt gtttatttcc gaatgcaaca
     3181 agctccgcat tacacccgaa catcactcca gatgagggct ttctgagtgt ggggtcaaat
     3241 agtttcatgt tccccaaatg gcccaaaact gacagtttaa acgctgtctt ggaacctaat
     3301 atgacaaaag cgtgatctca tccaagatga actaagtttg gttcgttgaa atgctaacgg
     3361 ccagttggtc aaaaagaaac ttccaaaagt cggcataccg tttgtcttgt ttggtattga
     3421 ttgacgaatg ctcaaaaata atctcattaa tgcttagcgc agtctctcta tcgcttctga
     3481 accccggtgc acctgtgccg aaacgcaaat ggggaaacac ccgctttttg gatgattatg
     3541 cattgtctcc acattgtatg cttccaagat tctggtggga atactgctga tagcctaacg
     3601 ttcatgatca aaatttaact gttctaaccc ctacttgaca gcaatatata aacagaagga
     3661 agctgccctg tcttaaacct ttttttttat catcattatt agcttacttt cataattgcg
     3721 actggttcca attgacaagc ttttgatttt aacgactttt aacgacaact tgagaagatc
     3781 aaaaaacaac taattattcg aaacgaggaa ttcatgtgtg atgaggaggt agccgccttg
     3841 gttgtggata acggttcagg tatgtgtaaa gcaggttttg ctggggacga cgcccctaga
     3901 gctgttttcc cttctattgt aggtcgaccc cgtcatcaag gcgtaatggt cggcatgggt
     3961 caaaaagact cttacgtggg agacgaagct caatctaaaa gaggaatatt gacattgaag
     4021 tacccaattg agcatggtat agttactaac tgggatgaca tggagaaaat ctggcatcac
     4081 acattctaca acgaattgag agtggcacct gaagagcatc cagtcttatt gacagaggct
     4141 cctcttaatc caaaagccaa tagagaaaaa atgacccaat tgatgttcga gactttcaac
     4201 actccagcca tgtatgtcgc catccaagct gtattgtcct tgtatgcttc aggcagaacc
     4261 acgggaatag ttttagactc aggagatggg gtctctcata ctgtaccaat atacgaaggt
     4321 tatgctttac ctcatgctat tttgcgtcta gatttggctg gccgtgacct gactgattac
     4381 cttatgaaaa ttcttacaga acgagggtac tcctttacca caactgctga gcgtgagatt
     4441 gtcagagata ttaaggaaaa attgtgttac gttgcccttg attttgaaca ggaaatggct
     4501 acagctgcat ctagtagttc acttgagaag tcttacgaat tgccagacgg tcaggttatt
     4561 accatcggca acgaaagatt tcgatgtcct gaggccctgt tccagccatc atttttggga
     4621 atggaggcat gtg
//
