LOCUS       sequence-362199-        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: I341L
FEATURES             Location/Qualifiers
     source          1..4633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      456..1044
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     945..964
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1119..1366)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     1332..1350
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(1432..1806)
                     /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(1825..1872)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(1879..2292)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      complement(2307..2553)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(2633..2650)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2666..2695)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(join(4069..4633,1..374))
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
ORIGIN
        1 atttgacccc acactcagaa agccctcatc tggagtgatg ttcgggtgta atgcggagct
       61 tgttgcattc ggaaataaac aaacatgaac ctcgccaggg gggccaggat agacaggcta
      121 ataaagtcat ggtgttagta gcctaataga aggaattgga atgagcgagc tccaatcaag
      181 cccaataact gggctggttt ttcgatggca aaagtgggtg ttgaggagaa gaggagtgga
      241 ggtcctgcgt ttgcaacggt ctgctgctag tgtatcccct cctgttgcgt ttggcactta
      301 tgtgtgagaa tggacctgtg gatgtcggat ggcaaaaagg tttcattcaa cctttcgtct
      361 ttggatgtta gatctgatct catgaccaaa atcccttaac gtgagttttc gttccactga
      421 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
      481 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
      541 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
      601 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
      661 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
      721 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
      781 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
      841 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
      901 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
      961 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     1021 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     1081 ttttgctggc cttttgctca catgttggtc tccagcttgc aaattaaagc cttcgagcgt
     1141 cccaaaacct tctcaagcaa ggttttcagt ataatgttac atgcgtacac gcgtctgtac
     1201 agaaaaaaaa gaaaaatttg aaatataaat aacgttctta atactaacat aactataaaa
     1261 aaataaatag ggacctagac ttcaggttgt ctaactcctt ccttttcggt tagagcggat
     1321 gtggggggag ggcgtgaatg taagcgtgac ataactaatt acatgatatc gacaaaggaa
     1381 aagggggacg gatctccgag gcctcggacc cgtcgggccg cgtcggacgt gtcagtcctg
     1441 ctcctcggcc acgaagtgca cgcagttgcc ggccgggtcg cgcagggcga actcccgccc
     1501 ccacggctgc tcgccgatct cggtcatggc cggcccggag gcgtcccgga agttcgtgga
     1561 cacgacctcc gaccactcgg cgtacagctc gtccaggccg cgcacccaca cccaggccag
     1621 ggtgttgtcc ggcaccacct ggtcctggac cgcgctgatg aacagggtca cgtcgtcccg
     1681 gaccacaccg gcgaagtcgt cctccacgaa gtcccgggag aacccgagcc ggtcggtcca
     1741 gaactcgacc gctccggcga cgtcgcgcgc ggtgagcacc ggaacggcac tggtcaactt
     1801 ggccatggtt tagttcctca ccttgtcgta ttatactatg ccgatatact atgccgatga
     1861 ttaattgtca acaccgccct tagattagat tgctatgctt tctttctaat gagcaagaag
     1921 taaaaaaagt tgtaatagaa caagaaaaat gaaactgaaa cttgagaaat tgaagaccgt
     1981 ttattaactt aaatatcaat gggaggtcat cgaaagagaa aaaaatcaaa aaaaaaaatt
     2041 ttcaagaaaa agaaacgtga taaaaatttt tattgccttt ttcgacgaag aaaaagaaac
     2101 gaggcggtct cttttttctt ttccaaacct ttagtacggg taattaacga caccctagag
     2161 gaagaaagag gggaaattta gtatgctgtg cttgggtgtt ttgaagtggt acggcgatgc
     2221 gcggagtccg agaaaatctg gaagagtaaa aaaggagtag aaacattttg aagctatggt
     2281 gtgtggggga tccgcacaaa cgaaggtctc acttaatctt ctgtactctg aagaggagtg
     2341 ggaaatacca agaaaaacat caaactcgaa tgattttccc aaacccctac cacaagatat
     2401 tcatcagctg cgagataggc tgatcaggag caagctcgta cgagaagaaa caaaatgaca
     2461 aaaaaaatcc tatactatat aggttacaaa taaaaaagta tcaaaaatga agcctgcatc
     2521 tctcaggcaa atggcattct gacatcctct tgattagaat ctagcaagac cggtcttctc
     2581 gtaagtgccc aacttgaact gaggaacagt catgtctaag gctacaaact caatgatgat
     2641 gatgatgatg gtcgacggcg ctattcagat cctcttctga gatgagtttt tgttctagaa
     2701 agctggcggc cgcactttca ccagcctgct tttcctgttc aatcgtttcc ttagatggca
     2761 atggattctt ctcttgggtc tcagtttttt tcaatttaga tttatcaaac ttctctatct
     2821 cagccatgtc gggcttatca gaggcagaac ctccacttga tcctccagat cctcctctag
     2881 aagcgccctg aaacaatact tcaagggatc caccactacc tccacttcca ccaaagcact
     2941 tacggtggac gatacttggt cctgattcat cgtattcctg tttagagatc cacatttgtt
     3001 ggaatgtact aagagaggcc aatatagaac cgcccaacca aacagaatac ttacgctcag
     3061 gtggtgcgat gattttaatt ttcatagtag atggagccaa ggccgtgatt tccttttgca
     3121 ttctatctgc aatacctggg tacatcgtag ttcctccact caggaccgta tttgcgtaca
     3181 agtcctttct aatagctgca gcacacttca tgatagaatt gtaagtagtc tcgtggattc
     3241 cacatgcctc cattcccaaa aatgatggct ggaacagggc ctcaggacat cgaaatcttt
     3301 cgttgccgat ggtaataacc tgaccgtctg gcaattcgta agacttctca agtgaactac
     3361 tagatgcagc tgtagccatt tcctgttcaa aatcaagggc aacgtaacac aatttttcct
     3421 taatatctct gacaatctca cgctcagcag ttgtggtaaa ggagtaccct cgttctgtaa
     3481 gaattttcat aaggtaatca gtcaggtcac ggccagccaa atctagacgc aaaatagcat
     3541 gaggtaaagc ataaccttcg tatattggta cagtatgaga gaccccatct cctgagtcta
     3601 aaactattcc cgtggttctg cctgaagcat acaaggacaa tacagcttgg atggcgacat
     3661 acatggctgg agtgttgaaa gtctcgaaca ttatttgggt cattttttct ctattggctt
     3721 ttggattaag aggagcctct gtcaataaga ctggatgctc ttcaggtgcc actctcaatt
     3781 cgttgtagaa tgtgtgatgc cagattttct ccatgtcatc ccagttagta actataccat
     3841 gctcaattgg gtacttcaat gtcaatattc ctcttttaga ttgagcttcg tctcccacgt
     3901 aagagtcttt ttgacccatg ccgaccatta cgccttgatg acggggtcga cctacaatag
     3961 aagggaaaac agctctaggg gcgtcgtccc cagcaaaacc tgctttacac atacctgaac
     4021 cgttatccac aaccaaggcg gctacctcct catcacacat gaattcctcg tttcgaataa
     4081 ttagttgttt tttgatcttc tcaagttgtc gttaaaagtc gttaaaatca aaagcttgtc
     4141 aattggaacc agtcgcaatt atgaaagtaa gctaataatg atgataaaaa aaaaggttta
     4201 agacagggca gcttccttct gtttatatat tgctgtcaag taggggttag aacagttaaa
     4261 ttttgatcat gaacgttagg ctatcagcag tattcccacc agaatcttgg aagcatacaa
     4321 tgtggagaca atgcataatc atccaaaaag cgggtgtttc cccatttgcg tttcggcaca
     4381 ggtgcaccgg ggttcagaag cgatagagag actgcgctaa gcattaatga gattattttt
     4441 gagcattcgt caatcaatac caaacaagac aaacggtatg ccgacttttg gaagtttctt
     4501 tttgaccaac tggccgttag catttcaacg aaccaaactt agttcatctt ggatgagatc
     4561 acgcttttgt catattaggt tccaagacag cgtttaaact gtcagttttg ggccatttgg
     4621 ggaacatgaa act
//
