LOCUS       sequence-362342-        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: I345L
FEATURES             Location/Qualifiers
     source          1..5978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             45..74
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             90..107
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      187..433
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        448..861
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        868..915
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             934..1308
                     /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      2635..2882
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2651..2669)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(2957..3545)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3037..3056)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3627..4565
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             4583..4600
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4850..4870
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 gtatcgtcca ccgtaagtgc ttttaagcgg ccgccagctt tctagaacaa aaactcatct
       61 cagaagagga tctgaatagc gccgtcgacc atcatcatca tcatcattga gtttgtagcc
      121 ttagacatga ctgttcctca gttcaagttg ggcacttacg agaagaccgg tcttgctaga
      181 ttctaatcaa gaggatgtca gaatgccatt tgcctgagag atgcaggctt catttttgat
      241 acttttttat ttgtaaccta tatagtatag gatttttttt gtcattttgt ttcttctcgt
      301 acgagcttgc tcctgatcag cctatctcgc agctgatgaa tatcttgtgg taggggtttg
      361 ggaaaatcat tcgagtttga tgtttttctt ggtatttccc actcctcttc agagtacaga
      421 agattaagtg agaccttcgt ttgtgcggat cccccacaca ccatagcttc aaaatgtttc
      481 tactcctttt ttactcttcc agattttctc ggactccgcg catcgccgta ccacttcaaa
      541 acacccaagc acagcatact aaatttcccc tctttcttcc tctagggtgt cgttaattac
      601 ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg tttctttttc ttcgtcgaaa
      661 aaggcaataa aaatttttat cacgtttctt tttcttgaaa attttttttt ttgatttttt
      721 tctctttcga tgacctccca ttgatattta agttaataaa cggtcttcaa tttctcaagt
      781 ttcagtttca tttttcttgt tctattacaa ctttttttac ttcttgctca ttagaaagaa
      841 agcatagcaa tctaatctaa gggcggtgtt gacaattaat catcggcata gtatatcggc
      901 atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg
      961 gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc
     1021 tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc
     1081 atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc
     1141 ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc
     1201 tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc
     1261 gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtccgacgc
     1321 ggcccgacgg gtccgatgat cttacccagc aatagtggac acgcggctaa gtgagtaaac
     1381 tctcagtcag aggtgactca catgacaaaa acagtatcaa ccagtaaaaa accccgtaaa
     1441 cagcattcgc ctgaatttcg cagtgaagcc ctgaagcttg ctgaacgcat cggtgttact
     1501 gccgcagccc gtgaactcag cctgtatgaa tcacagctct acaactggcg cagtaaacag
     1561 caaaatcagc agacgtcttc tgaacgtgaa ctggagatgt ctaccgagat tgcacgtctc
     1621 aaacgccagc tggcagaacg ggatgaagag ctggctatcc tccaaaaggc cgcgacatac
     1681 ttcgcgaagc gcctgaaatg aagtatgtct ttattgaaaa acatcaggct gagttcagca
     1741 tcaaagcaat gtgccgcgtg ctccgggtgg cccgcagcgg ctggtatacg tggtgtcagc
     1801 ggcggacaag gataagcacg cgtcagcagt tccgccaaca ctgcgacagc gttgtcctcg
     1861 cggcttttac ccggtcaaaa cagcgttacg gtgccccacg cctgacggat gaactgcgtg
     1921 ctcagggtta cccctttaac gtaaaaaccg tggcggcaag cctgcgccgt cagggactga
     1981 gggcaaaggc ctcccggaag ttcagcccgg tcagctaccg cgcacacggc ctgcctgtgt
     2041 cagaaaatct gttggagcag gatttttacg ccagtggccc gaaccagaag tgggcaggag
     2101 acatcacgta cttacgtaca gatgaaggct ggctgtatct ggcagtggtc attgacctgt
     2161 ggtcacgtgc cgttattggc tggtcaatgt cgccacgcat gacggcgcaa ctggcctgcg
     2221 atgccctgca gatggcgctg tggcggcgta agaggccccg gaacgttatc gttcacacgg
     2281 accgtggagg ccagtactgt tcagcagatt atcaggcgca actgaagcgg cataatctgc
     2341 gtggaagtat gagcgcaaaa ggttgctgct acgataatgc ctgcgtggaa agcttctttc
     2401 attcgctgaa agtggaatgt atccatggag aacactttat cagccgggaa ataatgcggg
     2461 caacggtgtt taattatatc gaatgtgatt acaatcggtg gcggcggcac agttggtgtg
     2521 gcggcctcag tccggaacaa tttgaaaaca agaacctcgc ttaggcctgt gtccatatta
     2581 cgtgggtagg atcacgaggc ctcggagatc cgtccccctt ttcctttgtc gatatcatgt
     2641 aattagttat gtcacgctta cattcacgcc ctccccccac atccgctcta accgaaaagg
     2701 aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt atgttagtat
     2761 taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc gtgtacgcat
     2821 gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag gctttaattt
     2881 gcaagctgga gaccaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2941 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3001 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3061 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3121 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3181 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3241 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3301 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3361 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3421 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3481 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3541 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3601 gttaagggat tttggtcatg agatcagatc taacatccaa agacgaaagg ttgaatgaaa
     3661 cctttttgcc atccgacatc cacaggtcca ttctcacaca taagtgccaa acgcaacagg
     3721 aggggataca ctagcagcag accgttgcaa acgcaggacc tccactcctc ttctcctcaa
     3781 cacccacttt tgccatcgaa aaaccagccc agttattggg cttgattgga gctcgctcat
     3841 tccaattcct tctattaggc tactaacacc atgactttat tagcctgtct atcctggccc
     3901 ccctggcgag gttcatgttt gtttatttcc gaatgcaaca agctccgcat tacacccgaa
     3961 catcactcca gatgagggct ttctgagtgt ggggtcaaat agtttcatgt tccccaaatg
     4021 gcccaaaact gacagtttaa acgctgtctt ggaacctaat atgacaaaag cgtgatctca
     4081 tccaagatga actaagtttg gttcgttgaa atgctaacgg ccagttggtc aaaaagaaac
     4141 ttccaaaagt cggcataccg tttgtcttgt ttggtattga ttgacgaatg ctcaaaaata
     4201 atctcattaa tgcttagcgc agtctctcta tcgcttctga accccggtgc acctgtgccg
     4261 aaacgcaaat ggggaaacac ccgctttttg gatgattatg cattgtctcc acattgtatg
     4321 cttccaagat tctggtggga atactgctga tagcctaacg ttcatgatca aaatttaact
     4381 gttctaaccc ctacttgaca gcaatatata aacagaagga agctgccctg tcttaaacct
     4441 ttttttttat catcattatt agcttacttt cataattgcg actggttcca attgacaagc
     4501 ttttgatttt aacgactttt aacgacaact tgagaagatc aaaaaacaac taattattcg
     4561 aaacgaggaa ttcgccatgg gtcaccatca tcatcatcac ggcggctccg gaggttctgg
     4621 tggatctgcc tccgacaagc ccgacatggc tgagattgag aagtttgaca aatccaagtt
     4681 aaaaaaaaca gagacccagg agaagaaccc tctacctagt aaggagacta ttgaacagga
     4741 aaaacaagcc ggtgaaagtg gtggatccgg tggctctggc ggttctggag gaagtggtgg
     4801 ttcaggagga tcaggtggat ctggcggatc cggaggaagt ggtggatcag aaaacttgta
     4861 ctttcaaggc catatgtgtg atgaggaggt agccgccttg gttgtggata acggttcagg
     4921 tatgtgtaaa gcaggttttg ctggggacga cgcccctaga gctgttttcc cttctattgt
     4981 aggtcgaccc cgtcatcaag gcgtaatggt cggcatgggt caaaaagact cttacgtggg
     5041 agacgaagct caatctaaaa gaggaatatt gacattgaag tacccaattg agcatggtat
     5101 agttactaac tgggatgaca tggagaaaat ctggcatcac acattctaca acgaattgag
     5161 agtggcacct gaagagcatc cagtcttatt gacagaggct cctcttaatc caaaagccaa
     5221 tagagaaaaa atgacccaaa taatgttcga gactttcaac actccagcca tgtatgtcgc
     5281 catccaagct gtattgtcct tgtatgcttc aggcagaacc acgggaatag ttttagactc
     5341 aggagatggg gtctctcata ctgtaccaat atacgaaggt tatgctttac ctcatgctat
     5401 tttgcgtcta gatttggctg gccgtgacct gactgattac cttatgaaaa ttcttacaga
     5461 acgagggtac tcctttacca caactgctga gcgtgagatt gtcagagata ttaaggaaaa
     5521 attgtgttac gttgcccttg attttgaaca ggaaatggct acagctgcat ctagtagttc
     5581 acttgagaag tcttacgaat tgccagacgg tcaggttatt accatcggca acgaaagatt
     5641 tcgatgtcct gaggccctgt tccagccatc atttttggga atggaggcat gtggaatcca
     5701 cgagactact tacaattcta tcatgaagtg tgctgcagct attagaaagg acttgtacgc
     5761 aaatacggtc ctgagtggag gaactacgat gtacccaggt attgcagata gaatgcaaaa
     5821 ggaaatcacg gccttggctc catctactat gaaaattaaa atcatcgcac cacctgagcg
     5881 taagtattct gtttggatag gcggttcttt gttggcctct cttagtacat tccaacaaat
     5941 gtggatctct aaacaggaat acgatgaatc aggaccaa
//
