LOCUS       sequence-362350-        5978 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 5978)
  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 5978)
  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: I369L
FEATURES             Location/Qualifiers
     source          1..5978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             339..368
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             384..401
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      481..727
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        742..1155
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        1162..1209
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1228..1602
                     /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      2929..3176
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2945..2963)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3251..3839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3331..3350)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3921..4859
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             4877..4894
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5144..5164
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 aggcatgtgg aatccacgag actacttaca attctatcat gaagtgtgct gcagctatta
       61 gaaaggactt gtacgcaaat acggtcctga gtggaggaac tacgatgtac ccaggtattg
      121 cagatagaat gcaaaaggaa atcacggcct tggctccatc tactatgaaa attaaaatca
      181 tcgcaccacc tgagcgtaag tattctgttt ggataggcgg ttctatattg gcctctctta
      241 gtacattcca acaaatgtgg atctctaaac aggaatacga tgaatcagga ccaagtttgg
      301 tccaccgtaa gtgcttttaa gcggccgcca gctttctaga acaaaaactc atctcagaag
      361 aggatctgaa tagcgccgtc gaccatcatc atcatcatca ttgagtttgt agccttagac
      421 atgactgttc ctcagttcaa gttgggcact tacgagaaga ccggtcttgc tagattctaa
      481 tcaagaggat gtcagaatgc catttgcctg agagatgcag gcttcatttt tgatactttt
      541 ttatttgtaa cctatatagt ataggatttt ttttgtcatt ttgtttcttc tcgtacgagc
      601 ttgctcctga tcagcctatc tcgcagctga tgaatatctt gtggtagggg tttgggaaaa
      661 tcattcgagt ttgatgtttt tcttggtatt tcccactcct cttcagagta cagaagatta
      721 agtgagacct tcgtttgtgc ggatccccca cacaccatag cttcaaaatg tttctactcc
      781 ttttttactc ttccagattt tctcggactc cgcgcatcgc cgtaccactt caaaacaccc
      841 aagcacagca tactaaattt cccctctttc ttcctctagg gtgtcgttaa ttacccgtac
      901 taaaggtttg gaaaagaaaa aagagaccgc ctcgtttctt tttcttcgtc gaaaaaggca
      961 ataaaaattt ttatcacgtt tctttttctt gaaaattttt ttttttgatt tttttctctt
     1021 tcgatgacct cccattgata tttaagttaa taaacggtct tcaatttctc aagtttcagt
     1081 ttcatttttc ttgttctatt acaacttttt ttacttcttg ctcattagaa agaaagcata
     1141 gcaatctaat ctaagggcgg tgttgacaat taatcatcgg catagtatat cggcatagta
     1201 taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc
     1261 accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg
     1321 gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc
     1381 gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg
     1441 gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg
     1501 ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg
     1561 gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtccg acgcggcccg
     1621 acgggtccga tgatcttacc cagcaatagt ggacacgcgg ctaagtgagt aaactctcag
     1681 tcagaggtga ctcacatgac aaaaacagta tcaaccagta aaaaaccccg taaacagcat
     1741 tcgcctgaat ttcgcagtga agccctgaag cttgctgaac gcatcggtgt tactgccgca
     1801 gcccgtgaac tcagcctgta tgaatcacag ctctacaact ggcgcagtaa acagcaaaat
     1861 cagcagacgt cttctgaacg tgaactggag atgtctaccg agattgcacg tctcaaacgc
     1921 cagctggcag aacgggatga agagctggct atcctccaaa aggccgcgac atacttcgcg
     1981 aagcgcctga aatgaagtat gtctttattg aaaaacatca ggctgagttc agcatcaaag
     2041 caatgtgccg cgtgctccgg gtggcccgca gcggctggta tacgtggtgt cagcggcgga
     2101 caaggataag cacgcgtcag cagttccgcc aacactgcga cagcgttgtc ctcgcggctt
     2161 ttacccggtc aaaacagcgt tacggtgccc cacgcctgac ggatgaactg cgtgctcagg
     2221 gttacccctt taacgtaaaa accgtggcgg caagcctgcg ccgtcaggga ctgagggcaa
     2281 aggcctcccg gaagttcagc ccggtcagct accgcgcaca cggcctgcct gtgtcagaaa
     2341 atctgttgga gcaggatttt tacgccagtg gcccgaacca gaagtgggca ggagacatca
     2401 cgtacttacg tacagatgaa ggctggctgt atctggcagt ggtcattgac ctgtggtcac
     2461 gtgccgttat tggctggtca atgtcgccac gcatgacggc gcaactggcc tgcgatgccc
     2521 tgcagatggc gctgtggcgg cgtaagaggc cccggaacgt tatcgttcac acggaccgtg
     2581 gaggccagta ctgttcagca gattatcagg cgcaactgaa gcggcataat ctgcgtggaa
     2641 gtatgagcgc aaaaggttgc tgctacgata atgcctgcgt ggaaagcttc tttcattcgc
     2701 tgaaagtgga atgtatccat ggagaacact ttatcagccg ggaaataatg cgggcaacgg
     2761 tgtttaatta tatcgaatgt gattacaatc ggtggcggcg gcacagttgg tgtggcggcc
     2821 tcagtccgga acaatttgaa aacaagaacc tcgcttaggc ctgtgtccat attacgtggg
     2881 taggatcacg aggcctcgga gatccgtccc ccttttcctt tgtcgatatc atgtaattag
     2941 ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag
     3001 ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa
     3061 cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca
     3121 ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcaagc
     3181 tggagaccaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3241 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3301 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3361 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3421 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3481 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3541 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3601 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3661 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3721 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3781 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3841 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3901 ggattttggt catgagatca gatctaacat ccaaagacga aaggttgaat gaaacctttt
     3961 tgccatccga catccacagg tccattctca cacataagtg ccaaacgcaa caggagggga
     4021 tacactagca gcagaccgtt gcaaacgcag gacctccact cctcttctcc tcaacaccca
     4081 cttttgccat cgaaaaacca gcccagttat tgggcttgat tggagctcgc tcattccaat
     4141 tccttctatt aggctactaa caccatgact ttattagcct gtctatcctg gcccccctgg
     4201 cgaggttcat gtttgtttat ttccgaatgc aacaagctcc gcattacacc cgaacatcac
     4261 tccagatgag ggctttctga gtgtggggtc aaatagtttc atgttcccca aatggcccaa
     4321 aactgacagt ttaaacgctg tcttggaacc taatatgaca aaagcgtgat ctcatccaag
     4381 atgaactaag tttggttcgt tgaaatgcta acggccagtt ggtcaaaaag aaacttccaa
     4441 aagtcggcat accgtttgtc ttgtttggta ttgattgacg aatgctcaaa aataatctca
     4501 ttaatgctta gcgcagtctc tctatcgctt ctgaaccccg gtgcacctgt gccgaaacgc
     4561 aaatggggaa acacccgctt tttggatgat tatgcattgt ctccacattg tatgcttcca
     4621 agattctggt gggaatactg ctgatagcct aacgttcatg atcaaaattt aactgttcta
     4681 acccctactt gacagcaata tataaacaga aggaagctgc cctgtcttaa accttttttt
     4741 ttatcatcat tattagctta ctttcataat tgcgactggt tccaattgac aagcttttga
     4801 ttttaacgac ttttaacgac aacttgagaa gatcaaaaaa caactaatta ttcgaaacga
     4861 ggaattcgcc atgggtcacc atcatcatca tcacggcggc tccggaggtt ctggtggatc
     4921 tgcctccgac aagcccgaca tggctgagat tgagaagttt gacaaatcca agttaaaaaa
     4981 aacagagacc caggagaaga accctctacc tagtaaggag actattgaac aggaaaaaca
     5041 agccggtgaa agtggtggat ccggtggctc tggcggttct ggaggaagtg gtggttcagg
     5101 aggatcaggt ggatctggcg gatccggagg aagtggtgga tcagaaaact tgtactttca
     5161 aggccatatg tgtgatgagg aggtagccgc cttggttgtg gataacggtt caggtatgtg
     5221 taaagcaggt tttgctgggg acgacgcccc tagagctgtt ttcccttcta ttgtaggtcg
     5281 accccgtcat caaggcgtaa tggtcggcat gggtcaaaaa gactcttacg tgggagacga
     5341 agctcaatct aaaagaggaa tattgacatt gaagtaccca attgagcatg gtatagttac
     5401 taactgggat gacatggaga aaatctggca tcacacattc tacaacgaat tgagagtggc
     5461 acctgaagag catccagtct tattgacaga ggctcctctt aatccaaaag ccaatagaga
     5521 aaaaatgacc caaataatgt tcgagacttt caacactcca gccatgtatg tcgccatcca
     5581 agctgtattg tccttgtatg cttcaggcag aaccacggga atagttttag actcaggaga
     5641 tggggtctct catactgtac caatatacga aggttatgct ttacctcatg ctattttgcg
     5701 tctagatttg gctggccgtg acctgactga ttaccttatg aaaattctta cagaacgagg
     5761 gtactccttt accacaactg ctgagcgtga gattgtcaga gatattaagg aaaaattgtg
     5821 ttacgttgcc cttgattttg aacaggaaat ggctacagct gcatctagta gttcacttga
     5881 gaagtcttac gaattgccag acggtcaggt tattaccatc ggcaacgaaa gatttcgatg
     5941 tcctgaggcc ctgttccagc catcattttt gggaatgg
//
