LOCUS       sequence-362187-        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: I64L
FEATURES             Location/Qualifiers
     source          1..5978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(901..921)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(1171..1188)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(1206..2144)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     rep_origin      2226..2814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2715..2734
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2889..3136)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     3102..3120
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(4463..4837)
                     /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(4856..4903)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(4910..5323)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      complement(5338..5584)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(5664..5681)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(5697..5726)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
ORIGIN
        1 gaccgtattt gcgtacaagt cctttctaat agctgcagca cacttcatga tagaattgta
       61 agtagtctcg tggattccac atgcctccat tcccaaaaat gatggctgga acagggcctc
      121 aggacatcga aatctttcgt tgccgatggt aataacctga ccgtctggca attcgtaaga
      181 cttctcaagt gaactactag atgcagctgt agccatttcc tgttcaaaat caagggcaac
      241 gtaacacaat ttttccttaa tatctctgac aatctcacgc tcagcagttg tggtaaagga
      301 gtaccctcgt tctgtaagaa ttttcataag gtaatcagtc aggtcacggc cagccaaatc
      361 tagacgcaaa atagcatgag gtaaagcata accttcgtat attggtacag tatgagagac
      421 cccatctcct gagtctaaaa ctattcccgt ggttctgcct gaagcataca aggacaatac
      481 agcttggatg gcgacataca tggctggagt gttgaaagtc tcgaacatta tttgggtcat
      541 tttttctcta ttggcttttg gattaagagg agcctctgtc aataagactg gatgctcttc
      601 aggtgccact ctcaattcgt tgtagaatgt gtgatgccag attttctcca tgtcatccca
      661 gttagtaact ataccatgct caattgggta cttcaatgtc aacaatcctc ttttagattg
      721 agcttcgtct cccacgtaag agtctttttg acccatgccg accattacgc cttgatgacg
      781 gggtcgacct acaatagaag ggaaaacagc tctaggggcg tcgtccccag caaaacctgc
      841 tttacacata cctgaaccgt tatccacaac caaggcggct acctcctcat cacacatatg
      901 gccttgaaag tacaagtttt ctgatccacc acttcctccg gatccgccag atccacctga
      961 tcctcctgaa ccaccacttc ctccagaacc gccagagcca ccggatccac cactttcacc
     1021 ggcttgtttt tcctgttcaa tagtctcctt actaggtaga gggttcttct cctgggtctc
     1081 tgtttttttt aacttggatt tgtcaaactt ctcaatctca gccatgtcgg gcttgtcgga
     1141 ggcagatcca ccagaacctc cggagccgcc gtgatgatga tgatggtgac ccatggcgaa
     1201 ttcctcgttt cgaataatta gttgtttttt gatcttctca agttgtcgtt aaaagtcgtt
     1261 aaaatcaaaa gcttgtcaat tggaaccagt cgcaattatg aaagtaagct aataatgatg
     1321 ataaaaaaaa aggtttaaga cagggcagct tccttctgtt tatatattgc tgtcaagtag
     1381 gggttagaac agttaaattt tgatcatgaa cgttaggcta tcagcagtat tcccaccaga
     1441 atcttggaag catacaatgt ggagacaatg cataatcatc caaaaagcgg gtgtttcccc
     1501 atttgcgttt cggcacaggt gcaccggggt tcagaagcga tagagagact gcgctaagca
     1561 ttaatgagat tatttttgag cattcgtcaa tcaataccaa acaagacaaa cggtatgccg
     1621 acttttggaa gtttcttttt gaccaactgg ccgttagcat ttcaacgaac caaacttagt
     1681 tcatcttgga tgagatcacg cttttgtcat attaggttcc aagacagcgt ttaaactgtc
     1741 agttttgggc catttgggga acatgaaact atttgacccc acactcagaa agccctcatc
     1801 tggagtgatg ttcgggtgta atgcggagct tgttgcattc ggaaataaac aaacatgaac
     1861 ctcgccaggg gggccaggat agacaggcta ataaagtcat ggtgttagta gcctaataga
     1921 aggaattgga atgagcgagc tccaatcaag cccaataact gggctggttt ttcgatggca
     1981 aaagtgggtg ttgaggagaa gaggagtgga ggtcctgcgt ttgcaacggt ctgctgctag
     2041 tgtatcccct cctgttgcgt ttggcactta tgtgtgagaa tggacctgtg gatgtcggat
     2101 ggcaaaaagg tttcattcaa cctttcgtct ttggatgtta gatctgatct catgaccaaa
     2161 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2221 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2281 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2341 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2401 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2461 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2521 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2581 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2641 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2701 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2761 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2821 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttggtc
     2881 tccagcttgc aaattaaagc cttcgagcgt cccaaaacct tctcaagcaa ggttttcagt
     2941 ataatgttac atgcgtacac gcgtctgtac agaaaaaaaa gaaaaatttg aaatataaat
     3001 aacgttctta atactaacat aactataaaa aaataaatag ggacctagac ttcaggttgt
     3061 ctaactcctt ccttttcggt tagagcggat gtggggggag ggcgtgaatg taagcgtgac
     3121 ataactaatt acatgatatc gacaaaggaa aagggggacg gatctccgag gcctcgtgat
     3181 cctacccacg taatatggac acaggcctaa gcgaggttct tgttttcaaa ttgttccgga
     3241 ctgaggccgc cacaccaact gtgccgccgc caccgattgt aatcacattc gatataatta
     3301 aacaccgttg cccgcattat ttcccggctg ataaagtgtt ctccatggat acattccact
     3361 ttcagcgaat gaaagaagct ttccacgcag gcattatcgt agcagcaacc ttttgcgctc
     3421 atacttccac gcagattatg ccgcttcagt tgcgcctgat aatctgctga acagtactgg
     3481 cctccacggt ccgtgtgaac gataacgttc cggggcctct tacgccgcca cagcgccatc
     3541 tgcagggcat cgcaggccag ttgcgccgtc atgcgtggcg acattgacca gccaataacg
     3601 gcacgtgacc acaggtcaat gaccactgcc agatacagcc agccttcatc tgtacgtaag
     3661 tacgtgatgt ctcctgccca cttctggttc gggccactgg cgtaaaaatc ctgctccaac
     3721 agattttctg acacaggcag gccgtgtgcg cggtagctga ccgggctgaa cttccgggag
     3781 gcctttgccc tcagtccctg acggcgcagg cttgccgcca cggtttttac gttaaagggg
     3841 taaccctgag cacgcagttc atccgtcagg cgtggggcac cgtaacgctg ttttgaccgg
     3901 gtaaaagccg cgaggacaac gctgtcgcag tgttggcgga actgctgacg cgtgcttatc
     3961 cttgtccgcc gctgacacca cgtataccag ccgctgcggg ccacccggag cacgcggcac
     4021 attgctttga tgctgaactc agcctgatgt ttttcaataa agacatactt catttcaggc
     4081 gcttcgcgaa gtatgtcgcg gccttttgga ggatagccag ctcttcatcc cgttctgcca
     4141 gctggcgttt gagacgtgca atctcggtag acatctccag ttcacgttca gaagacgtct
     4201 gctgattttg ctgtttactg cgccagttgt agagctgtga ttcatacagg ctgagttcac
     4261 gggctgcggc agtaacaccg atgcgttcag caagcttcag ggcttcactg cgaaattcag
     4321 gcgaatgctg tttacggggt tttttactgg ttgatactgt ttttgtcatg tgagtcacct
     4381 ctgactgaga gtttactcac ttagccgcgt gtccactatt gctgggtaag atcatcggac
     4441 ccgtcgggcc gcgtcggacg tgtcagtcct gctcctcggc cacgaagtgc acgcagttgc
     4501 cggccgggtc gcgcagggcg aactcccgcc cccacggctg ctcgccgatc tcggtcatgg
     4561 ccggcccgga ggcgtcccgg aagttcgtgg acacgacctc cgaccactcg gcgtacagct
     4621 cgtccaggcc gcgcacccac acccaggcca gggtgttgtc cggcaccacc tggtcctgga
     4681 ccgcgctgat gaacagggtc acgtcgtccc ggaccacacc ggcgaagtcg tcctccacga
     4741 agtcccggga gaacccgagc cggtcggtcc agaactcgac cgctccggcg acgtcgcgcg
     4801 cggtgagcac cggaacggca ctggtcaact tggccatggt ttagttcctc accttgtcgt
     4861 attatactat gccgatatac tatgccgatg attaattgtc aacaccgccc ttagattaga
     4921 ttgctatgct ttctttctaa tgagcaagaa gtaaaaaaag ttgtaataga acaagaaaaa
     4981 tgaaactgaa acttgagaaa ttgaagaccg tttattaact taaatatcaa tgggaggtca
     5041 tcgaaagaga aaaaaatcaa aaaaaaaaat tttcaagaaa aagaaacgtg ataaaaattt
     5101 ttattgcctt tttcgacgaa gaaaaagaaa cgaggcggtc tcttttttct tttccaaacc
     5161 tttagtacgg gtaattaacg acaccctaga ggaagaaaga ggggaaattt agtatgctgt
     5221 gcttgggtgt tttgaagtgg tacggcgatg cgcggagtcc gagaaaatct ggaagagtaa
     5281 aaaaggagta gaaacatttt gaagctatgg tgtgtggggg atccgcacaa acgaaggtct
     5341 cacttaatct tctgtactct gaagaggagt gggaaatacc aagaaaaaca tcaaactcga
     5401 atgattttcc caaaccccta ccacaagata ttcatcagct gcgagatagg ctgatcagga
     5461 gcaagctcgt acgagaagaa acaaaatgac aaaaaaaatc ctatactata taggttacaa
     5521 ataaaaaagt atcaaaaatg aagcctgcat ctctcaggca aatggcattc tgacatcctc
     5581 ttgattagaa tctagcaaga ccggtcttct cgtaagtgcc caacttgaac tgaggaacag
     5641 tcatgtctaa ggctacaaac tcaatgatga tgatgatgat ggtcgacggc gctattcaga
     5701 tcctcttctg agatgagttt ttgttctaga aagctggcgg ccgcttaaaa gcacttacgg
     5761 tggacgatac ttggtcctga ttcatcgtat tcctgtttag agatccacat ttgttggaat
     5821 gtactaagag aggccaatat agaaccgcct atccaaacag aatacttacg ctcaggtggt
     5881 gcgatgattt taattttcat agtagatgga gccaaggccg tgatttcctt ttgcattcta
     5941 tctgcaatac ctgggtacat cgtagttcct ccactcag
//
