LOCUS       sequence-362359-        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: I330L
FEATURES             Location/Qualifiers
     source          1..5978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             249..278
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             294..311
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      391..637
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        652..1065
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        1072..1119
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1138..1512
                     /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      2839..3086
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2855..2873)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3161..3749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3241..3260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3831..4769
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             4787..4804
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5054..5074
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 gtggaggaac tacgatgtac ccaggtattg cagatagaat gcaaaaggaa atcacggcct
       61 tggctccatc tactatgaaa attaaaatct tggcaccacc tgagcgtaag tattctgttt
      121 ggataggcgg ttctatattg gcctctctta gtacattcca acaaatgtgg atctctaaac
      181 aggaatacga tgaatcagga ccaagtatcg tccaccgtaa gtgcttttaa gcggccgcca
      241 gctttctaga acaaaaactc atctcagaag aggatctgaa tagcgccgtc gaccatcatc
      301 atcatcatca ttgagtttgt agccttagac atgactgttc ctcagttcaa gttgggcact
      361 tacgagaaga ccggtcttgc tagattctaa tcaagaggat gtcagaatgc catttgcctg
      421 agagatgcag gcttcatttt tgatactttt ttatttgtaa cctatatagt ataggatttt
      481 ttttgtcatt ttgtttcttc tcgtacgagc ttgctcctga tcagcctatc tcgcagctga
      541 tgaatatctt gtggtagggg tttgggaaaa tcattcgagt ttgatgtttt tcttggtatt
      601 tcccactcct cttcagagta cagaagatta agtgagacct tcgtttgtgc ggatccccca
      661 cacaccatag cttcaaaatg tttctactcc ttttttactc ttccagattt tctcggactc
      721 cgcgcatcgc cgtaccactt caaaacaccc aagcacagca tactaaattt cccctctttc
      781 ttcctctagg gtgtcgttaa ttacccgtac taaaggtttg gaaaagaaaa aagagaccgc
      841 ctcgtttctt tttcttcgtc gaaaaaggca ataaaaattt ttatcacgtt tctttttctt
      901 gaaaattttt ttttttgatt tttttctctt tcgatgacct cccattgata tttaagttaa
      961 taaacggtct tcaatttctc aagtttcagt ttcatttttc ttgttctatt acaacttttt
     1021 ttacttcttg ctcattagaa agaaagcata gcaatctaat ctaagggcgg tgttgacaat
     1081 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     1141 gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag
     1201 ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg
     1261 gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac
     1321 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc
     1381 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg
     1441 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag
     1501 gagcaggact gacacgtccg acgcggcccg acgggtccga tgatcttacc cagcaatagt
     1561 ggacacgcgg ctaagtgagt aaactctcag tcagaggtga ctcacatgac aaaaacagta
     1621 tcaaccagta aaaaaccccg taaacagcat tcgcctgaat ttcgcagtga agccctgaag
     1681 cttgctgaac gcatcggtgt tactgccgca gcccgtgaac tcagcctgta tgaatcacag
     1741 ctctacaact ggcgcagtaa acagcaaaat cagcagacgt cttctgaacg tgaactggag
     1801 atgtctaccg agattgcacg tctcaaacgc cagctggcag aacgggatga agagctggct
     1861 atcctccaaa aggccgcgac atacttcgcg aagcgcctga aatgaagtat gtctttattg
     1921 aaaaacatca ggctgagttc agcatcaaag caatgtgccg cgtgctccgg gtggcccgca
     1981 gcggctggta tacgtggtgt cagcggcgga caaggataag cacgcgtcag cagttccgcc
     2041 aacactgcga cagcgttgtc ctcgcggctt ttacccggtc aaaacagcgt tacggtgccc
     2101 cacgcctgac ggatgaactg cgtgctcagg gttacccctt taacgtaaaa accgtggcgg
     2161 caagcctgcg ccgtcaggga ctgagggcaa aggcctcccg gaagttcagc ccggtcagct
     2221 accgcgcaca cggcctgcct gtgtcagaaa atctgttgga gcaggatttt tacgccagtg
     2281 gcccgaacca gaagtgggca ggagacatca cgtacttacg tacagatgaa ggctggctgt
     2341 atctggcagt ggtcattgac ctgtggtcac gtgccgttat tggctggtca atgtcgccac
     2401 gcatgacggc gcaactggcc tgcgatgccc tgcagatggc gctgtggcgg cgtaagaggc
     2461 cccggaacgt tatcgttcac acggaccgtg gaggccagta ctgttcagca gattatcagg
     2521 cgcaactgaa gcggcataat ctgcgtggaa gtatgagcgc aaaaggttgc tgctacgata
     2581 atgcctgcgt ggaaagcttc tttcattcgc tgaaagtgga atgtatccat ggagaacact
     2641 ttatcagccg ggaaataatg cgggcaacgg tgtttaatta tatcgaatgt gattacaatc
     2701 ggtggcggcg gcacagttgg tgtggcggcc tcagtccgga acaatttgaa aacaagaacc
     2761 tcgcttaggc ctgtgtccat attacgtggg taggatcacg aggcctcgga gatccgtccc
     2821 ccttttcctt tgtcgatatc atgtaattag ttatgtcacg cttacattca cgccctcccc
     2881 ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt
     2941 atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     3001 ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt
     3061 tgggacgctc gaaggcttta atttgcaagc tggagaccaa catgtgagca aaaggccagc
     3121 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3181 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3241 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3301 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3361 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3421 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3481 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3541 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3601 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3661 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3721 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3781 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatca gatctaacat
     3841 ccaaagacga aaggttgaat gaaacctttt tgccatccga catccacagg tccattctca
     3901 cacataagtg ccaaacgcaa caggagggga tacactagca gcagaccgtt gcaaacgcag
     3961 gacctccact cctcttctcc tcaacaccca cttttgccat cgaaaaacca gcccagttat
     4021 tgggcttgat tggagctcgc tcattccaat tccttctatt aggctactaa caccatgact
     4081 ttattagcct gtctatcctg gcccccctgg cgaggttcat gtttgtttat ttccgaatgc
     4141 aacaagctcc gcattacacc cgaacatcac tccagatgag ggctttctga gtgtggggtc
     4201 aaatagtttc atgttcccca aatggcccaa aactgacagt ttaaacgctg tcttggaacc
     4261 taatatgaca aaagcgtgat ctcatccaag atgaactaag tttggttcgt tgaaatgcta
     4321 acggccagtt ggtcaaaaag aaacttccaa aagtcggcat accgtttgtc ttgtttggta
     4381 ttgattgacg aatgctcaaa aataatctca ttaatgctta gcgcagtctc tctatcgctt
     4441 ctgaaccccg gtgcacctgt gccgaaacgc aaatggggaa acacccgctt tttggatgat
     4501 tatgcattgt ctccacattg tatgcttcca agattctggt gggaatactg ctgatagcct
     4561 aacgttcatg atcaaaattt aactgttcta acccctactt gacagcaata tataaacaga
     4621 aggaagctgc cctgtcttaa accttttttt ttatcatcat tattagctta ctttcataat
     4681 tgcgactggt tccaattgac aagcttttga ttttaacgac ttttaacgac aacttgagaa
     4741 gatcaaaaaa caactaatta ttcgaaacga ggaattcgcc atgggtcacc atcatcatca
     4801 tcacggcggc tccggaggtt ctggtggatc tgcctccgac aagcccgaca tggctgagat
     4861 tgagaagttt gacaaatcca agttaaaaaa aacagagacc caggagaaga accctctacc
     4921 tagtaaggag actattgaac aggaaaaaca agccggtgaa agtggtggat ccggtggctc
     4981 tggcggttct ggaggaagtg gtggttcagg aggatcaggt ggatctggcg gatccggagg
     5041 aagtggtgga tcagaaaact tgtactttca aggccatatg tgtgatgagg aggtagccgc
     5101 cttggttgtg gataacggtt caggtatgtg taaagcaggt tttgctgggg acgacgcccc
     5161 tagagctgtt ttcccttcta ttgtaggtcg accccgtcat caaggcgtaa tggtcggcat
     5221 gggtcaaaaa gactcttacg tgggagacga agctcaatct aaaagaggaa tattgacatt
     5281 gaagtaccca attgagcatg gtatagttac taactgggat gacatggaga aaatctggca
     5341 tcacacattc tacaacgaat tgagagtggc acctgaagag catccagtct tattgacaga
     5401 ggctcctctt aatccaaaag ccaatagaga aaaaatgacc caaataatgt tcgagacttt
     5461 caacactcca gccatgtatg tcgccatcca agctgtattg tccttgtatg cttcaggcag
     5521 aaccacggga atagttttag actcaggaga tggggtctct catactgtac caatatacga
     5581 aggttatgct ttacctcatg ctattttgcg tctagatttg gctggccgtg acctgactga
     5641 ttaccttatg aaaattctta cagaacgagg gtactccttt accacaactg ctgagcgtga
     5701 gattgtcaga gatattaagg aaaaattgtg ttacgttgcc cttgattttg aacaggaaat
     5761 ggctacagct gcatctagta gttcacttga gaagtcttac gaattgccag acggtcaggt
     5821 tattaccatc ggcaacgaaa gatttcgatg tcctgaggcc ctgttccagc catcattttt
     5881 gggaatggag gcatgtggaa tccacgagac tacttacaat tctatcatga agtgtgctgc
     5941 agctattaga aaggacttgt acgcaaatac ggtcctga
//
