LOCUS       sequence-362293-        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: I329L
FEATURES             Location/Qualifiers
     source          1..4633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             480..509
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             525..542
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      622..868
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        883..1296
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        1303..1350
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1369..1743
                     /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      1809..2056
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1825..1843)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(2131..2719)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2211..2230)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2801..3739
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
ORIGIN
        1 caaatacggt cctgagtgga ggaactacga tgtacccagg tattgcagat agaatgcaaa
       61 aggaaatcac ggccttggct ccatctacta tgaaaattaa attgatcgca ccacctgagc
      121 gtaagtattc tgtttggata ggcggttcta tattggcctc tcttagtaca ttccaacaaa
      181 tgtggatctc taaacaggaa tacgatgaat caggaccaag tatcgtccac cgtaagtgct
      241 ttggtggaag tggaggtagt ggtggatccc ttgaagtatt gtttcagggc gcttctagag
      301 gaggatctgg aggatcaagt ggaggttctg cctctgataa gcccgacatg gctgagatag
      361 agaagtttga taaatctaaa ttgaaaaaaa ctgagaccca agagaagaat ccattgccat
      421 ctaaggaaac gattgaacag gaaaagcagg ctggtgaaag tgcggccgcc agctttctag
      481 aacaaaaact catctcagaa gaggatctga atagcgccgt cgaccatcat catcatcatc
      541 attgagtttg tagccttaga catgactgtt cctcagttca agttgggcac ttacgagaag
      601 accggtcttg ctagattcta atcaagagga tgtcagaatg ccatttgcct gagagatgca
      661 ggcttcattt ttgatacttt tttatttgta acctatatag tataggattt tttttgtcat
      721 tttgtttctt ctcgtacgag cttgctcctg atcagcctat ctcgcagctg atgaatatct
      781 tgtggtaggg gtttgggaaa atcattcgag tttgatgttt ttcttggtat ttcccactcc
      841 tcttcagagt acagaagatt aagtgagacc ttcgtttgtg cggatccccc acacaccata
      901 gcttcaaaat gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg
      961 ccgtaccact tcaaaacacc caagcacagc atactaaatt tcccctcttt cttcctctag
     1021 ggtgtcgtta attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct
     1081 ttttcttcgt cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaaatttt
     1141 tttttttgat ttttttctct ttcgatgacc tcccattgat atttaagtta ataaacggtc
     1201 ttcaatttct caagtttcag tttcattttt cttgttctat tacaactttt tttacttctt
     1261 gctcattaga aagaaagcat agcaatctaa tctaagggcg gtgttgacaa ttaatcatcg
     1321 gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg
     1381 accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc
     1441 gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac
     1501 gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc
     1561 tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg
     1621 aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg
     1681 gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac
     1741 tgacacgtcc gacgcggccc gacgggtccg aggcctcgga gatccgtccc ccttttcctt
     1801 tgtcgatatc atgtaattag ttatgtcacg cttacattca cgccctcccc ccacatccgc
     1861 tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat
     1921 agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacag
     1981 acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc
     2041 gaaggcttta atttgcaagc tggagaccaa catgtgagca aaaggccagc aaaaggccag
     2101 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2161 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2221 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2281 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2341 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2401 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2461 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2521 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2581 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2641 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2701 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2761 gaacgaaaac tcacgttaag ggattttggt catgagatca gatctaacat ccaaagacga
     2821 aaggttgaat gaaacctttt tgccatccga catccacagg tccattctca cacataagtg
     2881 ccaaacgcaa caggagggga tacactagca gcagaccgtt gcaaacgcag gacctccact
     2941 cctcttctcc tcaacaccca cttttgccat cgaaaaacca gcccagttat tgggcttgat
     3001 tggagctcgc tcattccaat tccttctatt aggctactaa caccatgact ttattagcct
     3061 gtctatcctg gcccccctgg cgaggttcat gtttgtttat ttccgaatgc aacaagctcc
     3121 gcattacacc cgaacatcac tccagatgag ggctttctga gtgtggggtc aaatagtttc
     3181 atgttcccca aatggcccaa aactgacagt ttaaacgctg tcttggaacc taatatgaca
     3241 aaagcgtgat ctcatccaag atgaactaag tttggttcgt tgaaatgcta acggccagtt
     3301 ggtcaaaaag aaacttccaa aagtcggcat accgtttgtc ttgtttggta ttgattgacg
     3361 aatgctcaaa aataatctca ttaatgctta gcgcagtctc tctatcgctt ctgaaccccg
     3421 gtgcacctgt gccgaaacgc aaatggggaa acacccgctt tttggatgat tatgcattgt
     3481 ctccacattg tatgcttcca agattctggt gggaatactg ctgatagcct aacgttcatg
     3541 atcaaaattt aactgttcta acccctactt gacagcaata tataaacaga aggaagctgc
     3601 cctgtcttaa accttttttt ttatcatcat tattagctta ctttcataat tgcgactggt
     3661 tccaattgac aagcttttga ttttaacgac ttttaacgac aacttgagaa gatcaaaaaa
     3721 caactaatta ttcgaaacga ggaattcatg tgtgatgagg aggtagccgc cttggttgtg
     3781 gataacggtt caggtatgtg taaagcaggt tttgctgggg acgacgcccc tagagctgtt
     3841 ttcccttcta ttgtaggtcg accccgtcat caaggcgtaa tggtcggcat gggtcaaaaa
     3901 gactcttacg tgggagacga agctcaatct aaaagaggaa tattgacatt gaagtaccca
     3961 attgagcatg gtatagttac taactgggat gacatggaga aaatctggca tcacacattc
     4021 tacaacgaat tgagagtggc acctgaagag catccagtct tattgacaga ggctcctctt
     4081 aatccaaaag ccaatagaga aaaaatgacc caaataatgt tcgagacttt caacactcca
     4141 gccatgtatg tcgccatcca agctgtattg tccttgtatg cttcaggcag aaccacggga
     4201 atagttttag actcaggaga tggggtctct catactgtac caatatacga aggttatgct
     4261 ttacctcatg ctattttgcg tctagatttg gctggccgtg acctgactga ttaccttatg
     4321 aaaattctta cagaacgagg gtactccttt accacaactg ctgagcgtga gattgtcaga
     4381 gatattaagg aaaaattgtg ttacgttgcc cttgattttg aacaggaaat ggctacagct
     4441 gcatctagta gttcacttga gaagtcttac gaattgccag acggtcaggt tattaccatc
     4501 ggcaacgaaa gatttcgatg tcctgaggcc ctgttccagc catcattttt gggaatggag
     4561 gcatgtggaa tccacgagac tacttacaat tctatcatga agtgtgctgc agctattaga
     4621 aaggacttgt acg
//
