LOCUS       sequence-362185-        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: I192L
FEATURES             Location/Qualifiers
     source          1..4633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(98..115)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(131..160)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(1534..2472)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     rep_origin      2554..3142
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3043..3062
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3217..3464)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     3430..3448
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(3530..3904)
                     /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(3923..3970)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(3977..4390)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      complement(join(4405..4633,1..18))
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
ORIGIN
        1 attctgacat cctcttgatt agaatctagc aagaccggtc ttctcgtaag tgcccaactt
       61 gaactgagga acagtcatgt ctaaggctac aaactcaatg atgatgatga tgatggtcga
      121 cggcgctatt cagatcctct tctgagatga gtttttgttc tagaaagctg gcggccgcac
      181 tttcaccagc ctgcttttcc tgttcaatcg tttccttaga tggcaatgga ttcttctctt
      241 gggtctcagt ttttttcaat ttagatttat caaacttctc tatctcagcc atgtcgggct
      301 tatcagaggc agaacctcca cttgatcctc cagatcctcc tctagaagcg ccctgaaaca
      361 atacttcaag ggatccacca ctacctccac ttccaccaaa gcacttacgg tggacgatac
      421 ttggtcctga ttcatcgtat tcctgtttag agatccacat ttgttggaat gtactaagag
      481 aggccaatat agaaccgcct atccaaacag aatacttacg ctcaggtggt gcgatgattt
      541 taattttcat agtagatgga gccaaggccg tgatttcctt ttgcattcta tctgcaatac
      601 ctgggtacat cgtagttcct ccactcagga ccgtatttgc gtacaagtcc tttctaatag
      661 ctgcagcaca cttcatgata gaattgtaag tagtctcgtg gattccacat gcctccattc
      721 ccaaaaatga tggctggaac agggcctcag gacatcgaaa tctttcgttg ccgatggtaa
      781 taacctgacc gtctggcaat tcgtaagact tctcaagtga actactagat gcagctgtag
      841 ccatttcctg ttcaaaatca agggcaacgt aacacaattt ttccttaata tctctgacaa
      901 tctcacgctc agcagttgtg gtaaaggagt accctcgttc tgtaagcaat ttcataaggt
      961 aatcagtcag gtcacggcca gccaaatcta gacgcaaaat agcatgaggt aaagcataac
     1021 cttcgtatat tggtacagta tgagagaccc catctcctga gtctaaaact attcccgtgg
     1081 ttctgcctga agcatacaag gacaatacag cttggatggc gacatacatg gctggagtgt
     1141 tgaaagtctc gaacattatt tgggtcattt tttctctatt ggcttttgga ttaagaggag
     1201 cctctgtcaa taagactgga tgctcttcag gtgccactct caattcgttg tagaatgtgt
     1261 gatgccagat tttctccatg tcatcccagt tagtaactat accatgctca attgggtact
     1321 tcaatgtcaa tattcctctt ttagattgag cttcgtctcc cacgtaagag tctttttgac
     1381 ccatgccgac cattacgcct tgatgacggg gtcgacctac aatagaaggg aaaacagctc
     1441 taggggcgtc gtccccagca aaacctgctt tacacatacc tgaaccgtta tccacaacca
     1501 aggcggctac ctcctcatca cacatgaatt cctcgtttcg aataattagt tgttttttga
     1561 tcttctcaag ttgtcgttaa aagtcgttaa aatcaaaagc ttgtcaattg gaaccagtcg
     1621 caattatgaa agtaagctaa taatgatgat aaaaaaaaag gtttaagaca gggcagcttc
     1681 cttctgttta tatattgctg tcaagtaggg gttagaacag ttaaattttg atcatgaacg
     1741 ttaggctatc agcagtattc ccaccagaat cttggaagca tacaatgtgg agacaatgca
     1801 taatcatcca aaaagcgggt gtttccccat ttgcgtttcg gcacaggtgc accggggttc
     1861 agaagcgata gagagactgc gctaagcatt aatgagatta tttttgagca ttcgtcaatc
     1921 aataccaaac aagacaaacg gtatgccgac ttttggaagt ttctttttga ccaactggcc
     1981 gttagcattt caacgaacca aacttagttc atcttggatg agatcacgct tttgtcatat
     2041 taggttccaa gacagcgttt aaactgtcag ttttgggcca tttggggaac atgaaactat
     2101 ttgaccccac actcagaaag ccctcatctg gagtgatgtt cgggtgtaat gcggagcttg
     2161 ttgcattcgg aaataaacaa acatgaacct cgccaggggg gccaggatag acaggctaat
     2221 aaagtcatgg tgttagtagc ctaatagaag gaattggaat gagcgagctc caatcaagcc
     2281 caataactgg gctggttttt cgatggcaaa agtgggtgtt gaggagaaga ggagtggagg
     2341 tcctgcgttt gcaacggtct gctgctagtg tatcccctcc tgttgcgttt ggcacttatg
     2401 tgtgagaatg gacctgtgga tgtcggatgg caaaaaggtt tcattcaacc tttcgtcttt
     2461 ggatgttaga tctgatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     2521 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2581 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2641 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2701 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2761 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2821 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2881 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2941 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3001 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3061 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3121 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3181 ttgctggcct tttgctcaca tgttggtctc cagcttgcaa attaaagcct tcgagcgtcc
     3241 caaaaccttc tcaagcaagg ttttcagtat aatgttacat gcgtacacgc gtctgtacag
     3301 aaaaaaaaga aaaatttgaa atataaataa cgttcttaat actaacataa ctataaaaaa
     3361 ataaataggg acctagactt caggttgtct aactccttcc ttttcggtta gagcggatgt
     3421 ggggggaggg cgtgaatgta agcgtgacat aactaattac atgatatcga caaaggaaaa
     3481 gggggacgga tctccgaggc ctcggacccg tcgggccgcg tcggacgtgt cagtcctgct
     3541 cctcggccac gaagtgcacg cagttgccgg ccgggtcgcg cagggcgaac tcccgccccc
     3601 acggctgctc gccgatctcg gtcatggccg gcccggaggc gtcccggaag ttcgtggaca
     3661 cgacctccga ccactcggcg tacagctcgt ccaggccgcg cacccacacc caggccaggg
     3721 tgttgtccgg caccacctgg tcctggaccg cgctgatgaa cagggtcacg tcgtcccgga
     3781 ccacaccggc gaagtcgtcc tccacgaagt cccgggagaa cccgagccgg tcggtccaga
     3841 actcgaccgc tccggcgacg tcgcgcgcgg tgagcaccgg aacggcactg gtcaacttgg
     3901 ccatggttta gttcctcacc ttgtcgtatt atactatgcc gatatactat gccgatgatt
     3961 aattgtcaac accgccctta gattagattg ctatgctttc tttctaatga gcaagaagta
     4021 aaaaaagttg taatagaaca agaaaaatga aactgaaact tgagaaattg aagaccgttt
     4081 attaacttaa atatcaatgg gaggtcatcg aaagagaaaa aaatcaaaaa aaaaaatttt
     4141 caagaaaaag aaacgtgata aaaattttta ttgccttttt cgacgaagaa aaagaaacga
     4201 ggcggtctct tttttctttt ccaaaccttt agtacgggta attaacgaca ccctagagga
     4261 agaaagaggg gaaatttagt atgctgtgct tgggtgtttt gaagtggtac ggcgatgcgc
     4321 ggagtccgag aaaatctgga agagtaaaaa aggagtagaa acattttgaa gctatggtgt
     4381 gtgggggatc cgcacaaacg aaggtctcac ttaatcttct gtactctgaa gaggagtggg
     4441 aaataccaag aaaaacatca aactcgaatg attttcccaa acccctacca caagatattc
     4501 atcagctgcg agataggctg atcaggagca agctcgtacg agaagaaaca aaatgacaaa
     4561 aaaaatccta tactatatag gttacaaata aaaaagtatc aaaaatgaag cctgcatctc
     4621 tcaggcaaat ggc
//
