LOCUS       sequence-362387-        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: K118N
FEATURES             Location/Qualifiers
     source          1..5978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             413..442
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             458..475
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      555..801
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        816..1229
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        1236..1283
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1302..1676
                     /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      3003..3250
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3019..3037)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3325..3913)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3405..3424)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3995..4933
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             4951..4968
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5218..5238
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 tcaggttatt accatcggca acgaaagatt tcgatgtcct gaggccctgt tccagccatc
       61 atttttggga atggaggcat gtggaatcca cgagactact tacaattcta tcatgaagtg
      121 tgctgcagct attagaaagg acttgtacgc aaatacggtc ctgagtggag gaactacgat
      181 gtacccaggt attgcagata gaatgcaaaa ggaaatcacg gccttggctc catctactat
      241 gaaaattaaa atcatcgcac cacctgagcg taagtattct gtttggatag gcggttctat
      301 attggcctct cttagtacat tccaacaaat gtggatctct aaacaggaat acgatgaatc
      361 aggaccaagt atcgtccacc gtaagtgctt ttaagcggcc gccagctttc tagaacaaaa
      421 actcatctca gaagaggatc tgaatagcgc cgtcgaccat catcatcatc atcattgagt
      481 ttgtagcctt agacatgact gttcctcagt tcaagttggg cacttacgag aagaccggtc
      541 ttgctagatt ctaatcaaga ggatgtcaga atgccatttg cctgagagat gcaggcttca
      601 tttttgatac ttttttattt gtaacctata tagtatagga ttttttttgt cattttgttt
      661 cttctcgtac gagcttgctc ctgatcagcc tatctcgcag ctgatgaata tcttgtggta
      721 ggggtttggg aaaatcattc gagtttgatg tttttcttgg tatttcccac tcctcttcag
      781 agtacagaag attaagtgag accttcgttt gtgcggatcc cccacacacc atagcttcaa
      841 aatgtttcta ctcctttttt actcttccag attttctcgg actccgcgca tcgccgtacc
      901 acttcaaaac acccaagcac agcatactaa atttcccctc tttcttcctc tagggtgtcg
      961 ttaattaccc gtactaaagg tttggaaaag aaaaaagaga ccgcctcgtt tctttttctt
     1021 cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt tcttgaaaat tttttttttt
     1081 gatttttttc tctttcgatg acctcccatt gatatttaag ttaataaacg gtcttcaatt
     1141 tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt cttgctcatt
     1201 agaaagaaag catagcaatc taatctaagg gcggtgttga caattaatca tcggcatagt
     1261 atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag ttgaccagtg
     1321 ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc
     1381 tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga
     1441 ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt
     1501 gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc
     1561 gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg
     1621 ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg
     1681 tccgacgcgg cccgacgggt ccgatgatct tacccagcaa tagtggacac gcggctaagt
     1741 gagtaaactc tcagtcagag gtgactcaca tgacaaaaac agtatcaacc agtaaaaaac
     1801 cccgtaaaca gcattcgcct gaatttcgca gtgaagccct gaagcttgct gaacgcatcg
     1861 gtgttactgc cgcagcccgt gaactcagcc tgtatgaatc acagctctac aactggcgca
     1921 gtaaacagca aaatcagcag acgtcttctg aacgtgaact ggagatgtct accgagattg
     1981 cacgtctcaa acgccagctg gcagaacggg atgaagagct ggctatcctc caaaaggccg
     2041 cgacatactt cgcgaagcgc ctgaaatgaa gtatgtcttt attgaaaaac atcaggctga
     2101 gttcagcatc aaagcaatgt gccgcgtgct ccgggtggcc cgcagcggct ggtatacgtg
     2161 gtgtcagcgg cggacaagga taagcacgcg tcagcagttc cgccaacact gcgacagcgt
     2221 tgtcctcgcg gcttttaccc ggtcaaaaca gcgttacggt gccccacgcc tgacggatga
     2281 actgcgtgct cagggttacc cctttaacgt aaaaaccgtg gcggcaagcc tgcgccgtca
     2341 gggactgagg gcaaaggcct cccggaagtt cagcccggtc agctaccgcg cacacggcct
     2401 gcctgtgtca gaaaatctgt tggagcagga tttttacgcc agtggcccga accagaagtg
     2461 ggcaggagac atcacgtact tacgtacaga tgaaggctgg ctgtatctgg cagtggtcat
     2521 tgacctgtgg tcacgtgccg ttattggctg gtcaatgtcg ccacgcatga cggcgcaact
     2581 ggcctgcgat gccctgcaga tggcgctgtg gcggcgtaag aggccccgga acgttatcgt
     2641 tcacacggac cgtggaggcc agtactgttc agcagattat caggcgcaac tgaagcggca
     2701 taatctgcgt ggaagtatga gcgcaaaagg ttgctgctac gataatgcct gcgtggaaag
     2761 cttctttcat tcgctgaaag tggaatgtat ccatggagaa cactttatca gccgggaaat
     2821 aatgcgggca acggtgttta attatatcga atgtgattac aatcggtggc ggcggcacag
     2881 ttggtgtggc ggcctcagtc cggaacaatt tgaaaacaag aacctcgctt aggcctgtgt
     2941 ccatattacg tgggtaggat cacgaggcct cggagatccg tccccctttt cctttgtcga
     3001 tatcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac
     3061 cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat
     3121 gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt
     3181 gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc
     3241 tttaatttgc aagctggaga ccaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3301 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3361 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3421 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3481 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3541 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3601 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3661 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3721 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3781 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3841 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3901 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3961 aaactcacgt taagggattt tggtcatgag atcagatcta acatccaaag acgaaaggtt
     4021 gaatgaaacc tttttgccat ccgacatcca caggtccatt ctcacacata agtgccaaac
     4081 gcaacaggag gggatacact agcagcagac cgttgcaaac gcaggacctc cactcctctt
     4141 ctcctcaaca cccacttttg ccatcgaaaa accagcccag ttattgggct tgattggagc
     4201 tcgctcattc caattccttc tattaggcta ctaacaccat gactttatta gcctgtctat
     4261 cctggccccc ctggcgaggt tcatgtttgt ttatttccga atgcaacaag ctccgcatta
     4321 cacccgaaca tcactccaga tgagggcttt ctgagtgtgg ggtcaaatag tttcatgttc
     4381 cccaaatggc ccaaaactga cagtttaaac gctgtcttgg aacctaatat gacaaaagcg
     4441 tgatctcatc caagatgaac taagtttggt tcgttgaaat gctaacggcc agttggtcaa
     4501 aaagaaactt ccaaaagtcg gcataccgtt tgtcttgttt ggtattgatt gacgaatgct
     4561 caaaaataat ctcattaatg cttagcgcag tctctctatc gcttctgaac cccggtgcac
     4621 ctgtgccgaa acgcaaatgg ggaaacaccc gctttttgga tgattatgca ttgtctccac
     4681 attgtatgct tccaagattc tggtgggaat actgctgata gcctaacgtt catgatcaaa
     4741 atttaactgt tctaacccct acttgacagc aatatataaa cagaaggaag ctgccctgtc
     4801 ttaaaccttt ttttttatca tcattattag cttactttca taattgcgac tggttccaat
     4861 tgacaagctt ttgattttaa cgacttttaa cgacaacttg agaagatcaa aaaacaacta
     4921 attattcgaa acgaggaatt cgccatgggt caccatcatc atcatcacgg cggctccgga
     4981 ggttctggtg gatctgcctc cgacaagccc gacatggctg agattgagaa gtttgacaaa
     5041 tccaagttaa aaaaaacaga gacccaggag aagaaccctc tacctagtaa ggagactatt
     5101 gaacaggaaa aacaagccgg tgaaagtggt ggatccggtg gctctggcgg ttctggagga
     5161 agtggtggtt caggaggatc aggtggatct ggcggatccg gaggaagtgg tggatcagaa
     5221 aacttgtact ttcaaggcca tatgtgtgat gaggaggtag ccgccttggt tgtggataac
     5281 ggttcaggta tgtgtaaagc aggttttgct ggggacgacg cccctagagc tgttttccct
     5341 tctattgtag gtcgaccccg tcatcaaggc gtaatggtcg gcatgggtca aaaagactct
     5401 tacgtgggag acgaagctca atctaaaaga ggaatattga cattgaagta cccaattgag
     5461 catggtatag ttactaactg ggatgacatg gagaaaatct ggcatcacac attctacaac
     5521 gaattgagag tggcacctga agagcatcca gtcttattga cagaggctcc tcttaatcca
     5581 aaagccaata gagaaaacat gacccaaata atgttcgaga ctttcaacac tccagccatg
     5641 tatgtcgcca tccaagctgt attgtccttg tatgcttcag gcagaaccac gggaatagtt
     5701 ttagactcag gagatggggt ctctcatact gtaccaatat acgaaggtta tgctttacct
     5761 catgctattt tgcgtctaga tttggctggc cgtgacctga ctgattacct tatgaaaatt
     5821 cttacagaac gagggtactc ctttaccaca actgctgagc gtgagattgt cagagatatt
     5881 aaggaaaaat tgtgttacgt tgcccttgat tttgaacagg aaatggctac agctgcatct
     5941 agtagttcac ttgagaagtc ttacgaattg ccagacgg
//
