LOCUS       sequence-362345-        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: I85L
FEATURES             Location/Qualifiers
     source          1..4633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(172..418)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(498..515)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(531..560)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(1934..2872)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     rep_origin      2954..3542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3443..3462
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3617..3864)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     3830..3848
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(3930..4304)
                     /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(4323..4370)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(4377..4633,1..157))
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 acgggtaatt aacgacaccc tagaggaaga aagaggggaa atttagtatg ctgtgcttgg
       61 gtgttttgaa gtggtacggc gatgcgcgga gtccgagaaa atctggaaga gtaaaaaagg
      121 agtagaaaca ttttgaagct atggtgtgtg ggggatccgc acaaacgaag gtctcactta
      181 atcttctgta ctctgaagag gagtgggaaa taccaagaaa aacatcaaac tcgaatgatt
      241 ttcccaaacc cctaccacaa gatattcatc agctgcgaga taggctgatc aggagcaagc
      301 tcgtacgaga agaaacaaaa tgacaaaaaa aatcctatac tatataggtt acaaataaaa
      361 aagtatcaaa aatgaagcct gcatctctca ggcaaatggc attctgacat cctcttgatt
      421 agaatctagc aagaccggtc ttctcgtaag tgcccaactt gaactgagga acagtcatgt
      481 ctaaggctac aaactcaatg atgatgatga tgatggtcga cggcgctatt cagatcctct
      541 tctgagatga gtttttgttc tagaaagctg gcggccgcac tttcaccagc ctgcttttcc
      601 tgttcaatcg tttccttaga tggcaatgga ttcttctctt gggtctcagt ttttttcaat
      661 ttagatttat caaacttctc tatctcagcc atgtcgggct tatcagaggc agaacctcca
      721 cttgatcctc cagatcctcc tctagaagcg ccctgaaaca atacttcaag ggatccacca
      781 ctacctccac ttccaccaaa gcacttacgg tggacgatac ttggtcctga ttcatcgtat
      841 tcctgtttag agatccacat ttgttggaat gtactaagag aggccaatat agaaccgcct
      901 atccaaacag aatacttacg ctcaggtggt gcgatgattt taattttcat agtagatgga
      961 gccaaggccg tgatttcctt ttgcattcta tctgcaatac ctgggtacat cgtagttcct
     1021 ccactcagga ccgtatttgc gtacaagtcc tttctaatag ctgcagcaca cttcatgata
     1081 gaattgtaag tagtctcgtg gattccacat gcctccattc ccaaaaatga tggctggaac
     1141 agggcctcag gacatcgaaa tctttcgttg ccgatggtaa taacctgacc gtctggcaat
     1201 tcgtaagact tctcaagtga actactagat gcagctgtag ccatttcctg ttcaaaatca
     1261 agggcaacgt aacacaattt ttccttaata tctctgacaa tctcacgctc agcagttgtg
     1321 gtaaaggagt accctcgttc tgtaagaatt ttcataaggt aatcagtcag gtcacggcca
     1381 gccaaatcta gacgcaaaat agcatgaggt aaagcataac cttcgtatat tggtacagta
     1441 tgagagaccc catctcctga gtctaaaact attcccgtgg ttctgcctga agcatacaag
     1501 gacaatacag cttggatggc gacatacatg gctggagtgt tgaaagtctc gaacattatt
     1561 tgggtcattt tttctctatt ggcttttgga ttaagaggag cctctgtcaa taagactgga
     1621 tgctcttcag gtgccactct caattcgttg tagaatgtgt gatgccacaa tttctccatg
     1681 tcatcccagt tagtaactat accatgctca attgggtact tcaatgtcaa tattcctctt
     1741 ttagattgag cttcgtctcc cacgtaagag tctttttgac ccatgccgac cattacgcct
     1801 tgatgacggg gtcgacctac aatagaaggg aaaacagctc taggggcgtc gtccccagca
     1861 aaacctgctt tacacatacc tgaaccgtta tccacaacca aggcggctac ctcctcatca
     1921 cacatgaatt cctcgtttcg aataattagt tgttttttga tcttctcaag ttgtcgttaa
     1981 aagtcgttaa aatcaaaagc ttgtcaattg gaaccagtcg caattatgaa agtaagctaa
     2041 taatgatgat aaaaaaaaag gtttaagaca gggcagcttc cttctgttta tatattgctg
     2101 tcaagtaggg gttagaacag ttaaattttg atcatgaacg ttaggctatc agcagtattc
     2161 ccaccagaat cttggaagca tacaatgtgg agacaatgca taatcatcca aaaagcgggt
     2221 gtttccccat ttgcgtttcg gcacaggtgc accggggttc agaagcgata gagagactgc
     2281 gctaagcatt aatgagatta tttttgagca ttcgtcaatc aataccaaac aagacaaacg
     2341 gtatgccgac ttttggaagt ttctttttga ccaactggcc gttagcattt caacgaacca
     2401 aacttagttc atcttggatg agatcacgct tttgtcatat taggttccaa gacagcgttt
     2461 aaactgtcag ttttgggcca tttggggaac atgaaactat ttgaccccac actcagaaag
     2521 ccctcatctg gagtgatgtt cgggtgtaat gcggagcttg ttgcattcgg aaataaacaa
     2581 acatgaacct cgccaggggg gccaggatag acaggctaat aaagtcatgg tgttagtagc
     2641 ctaatagaag gaattggaat gagcgagctc caatcaagcc caataactgg gctggttttt
     2701 cgatggcaaa agtgggtgtt gaggagaaga ggagtggagg tcctgcgttt gcaacggtct
     2761 gctgctagtg tatcccctcc tgttgcgttt ggcacttatg tgtgagaatg gacctgtgga
     2821 tgtcggatgg caaaaaggtt tcattcaacc tttcgtcttt ggatgttaga tctgatctca
     2881 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2941 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3001 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3061 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     3121 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3181 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3241 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3301 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3361 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3421 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3481 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3541 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3601 tgttggtctc cagcttgcaa attaaagcct tcgagcgtcc caaaaccttc tcaagcaagg
     3661 ttttcagtat aatgttacat gcgtacacgc gtctgtacag aaaaaaaaga aaaatttgaa
     3721 atataaataa cgttcttaat actaacataa ctataaaaaa ataaataggg acctagactt
     3781 caggttgtct aactccttcc ttttcggtta gagcggatgt ggggggaggg cgtgaatgta
     3841 agcgtgacat aactaattac atgatatcga caaaggaaaa gggggacgga tctccgaggc
     3901 ctcggacccg tcgggccgcg tcggacgtgt cagtcctgct cctcggccac gaagtgcacg
     3961 cagttgccgg ccgggtcgcg cagggcgaac tcccgccccc acggctgctc gccgatctcg
     4021 gtcatggccg gcccggaggc gtcccggaag ttcgtggaca cgacctccga ccactcggcg
     4081 tacagctcgt ccaggccgcg cacccacacc caggccaggg tgttgtccgg caccacctgg
     4141 tcctggaccg cgctgatgaa cagggtcacg tcgtcccgga ccacaccggc gaagtcgtcc
     4201 tccacgaagt cccgggagaa cccgagccgg tcggtccaga actcgaccgc tccggcgacg
     4261 tcgcgcgcgg tgagcaccgg aacggcactg gtcaacttgg ccatggttta gttcctcacc
     4321 ttgtcgtatt atactatgcc gatatactat gccgatgatt aattgtcaac accgccctta
     4381 gattagattg ctatgctttc tttctaatga gcaagaagta aaaaaagttg taatagaaca
     4441 agaaaaatga aactgaaact tgagaaattg aagaccgttt attaacttaa atatcaatgg
     4501 gaggtcatcg aaagagaaaa aaatcaaaaa aaaaaatttt caagaaaaag aaacgtgata
     4561 aaaattttta ttgccttttt cgacgaagaa aaagaaacga ggcggtctct tttttctttt
     4621 ccaaaccttt agt
//
