LOCUS       sequence-362343-        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: I75V
FEATURES             Location/Qualifiers
     source          1..4633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(805..1743)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     rep_origin      1825..2413
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2314..2333
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2488..2735)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2701..2719
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(2801..3175)
                     /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(3194..3241)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(3248..3661)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      complement(3676..3922)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(4002..4019)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4035..4064)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
ORIGIN
        1 atggctggaa cagggcctca ggacatcgaa atctttcgtt gccgatggta ataacctgac
       61 cgtctggcaa ttcgtaagac ttctcaagtg aactactaga tgcagctgta gccatttcct
      121 gttcaaaatc aagggcaacg taacacaatt tttccttaat atctctgaca atctcacgct
      181 cagcagttgt ggtaaaggag taccctcgtt ctgtaagaat tttcataagg taatcagtca
      241 ggtcacggcc agccaaatct agacgcaaaa tagcatgagg taaagcataa ccttcgtata
      301 ttggtacagt atgagagacc ccatctcctg agtctaaaac tattcccgtg gttctgcctg
      361 aagcatacaa ggacaataca gcttggatgg cgacatacat ggctggagtg ttgaaagtct
      421 cgaacattat ttgggtcatt ttttctctat tggcttttgg attaagagga gcctctgtca
      481 ataagactgg atgctcttca ggtgccactc tcaattcgtt gtagaatgtg tgatgccaga
      541 ttttctccat gtcatcccag ttagtaacaa caccatgctc aattgggtac ttcaatgtca
      601 atattcctct tttagattga gcttcgtctc ccacgtaaga gtctttttga cccatgccga
      661 ccattacgcc ttgatgacgg ggtcgaccta caatagaagg gaaaacagct ctaggggcgt
      721 cgtccccagc aaaacctgct ttacacatac ctgaaccgtt atccacaacc aaggcggcta
      781 cctcctcatc acacatgaat tcctcgtttc gaataattag ttgttttttg atcttctcaa
      841 gttgtcgtta aaagtcgtta aaatcaaaag cttgtcaatt ggaaccagtc gcaattatga
      901 aagtaagcta ataatgatga taaaaaaaaa ggtttaagac agggcagctt ccttctgttt
      961 atatattgct gtcaagtagg ggttagaaca gttaaatttt gatcatgaac gttaggctat
     1021 cagcagtatt cccaccagaa tcttggaagc atacaatgtg gagacaatgc ataatcatcc
     1081 aaaaagcggg tgtttcccca tttgcgtttc ggcacaggtg caccggggtt cagaagcgat
     1141 agagagactg cgctaagcat taatgagatt atttttgagc attcgtcaat caataccaaa
     1201 caagacaaac ggtatgccga cttttggaag tttctttttg accaactggc cgttagcatt
     1261 tcaacgaacc aaacttagtt catcttggat gagatcacgc ttttgtcata ttaggttcca
     1321 agacagcgtt taaactgtca gttttgggcc atttggggaa catgaaacta tttgacccca
     1381 cactcagaaa gccctcatct ggagtgatgt tcgggtgtaa tgcggagctt gttgcattcg
     1441 gaaataaaca aacatgaacc tcgccagggg ggccaggata gacaggctaa taaagtcatg
     1501 gtgttagtag cctaatagaa ggaattggaa tgagcgagct ccaatcaagc ccaataactg
     1561 ggctggtttt tcgatggcaa aagtgggtgt tgaggagaag aggagtggag gtcctgcgtt
     1621 tgcaacggtc tgctgctagt gtatcccctc ctgttgcgtt tggcacttat gtgtgagaat
     1681 ggacctgtgg atgtcggatg gcaaaaaggt ttcattcaac ctttcgtctt tggatgttag
     1741 atctgatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1801 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1861 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1921 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     1981 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2041 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2101 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2161 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2221 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2281 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2341 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2401 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2461 ttttgctcac atgttggtct ccagcttgca aattaaagcc ttcgagcgtc ccaaaacctt
     2521 ctcaagcaag gttttcagta taatgttaca tgcgtacacg cgtctgtaca gaaaaaaaag
     2581 aaaaatttga aatataaata acgttcttaa tactaacata actataaaaa aataaatagg
     2641 gacctagact tcaggttgtc taactccttc cttttcggtt agagcggatg tggggggagg
     2701 gcgtgaatgt aagcgtgaca taactaatta catgatatcg acaaaggaaa agggggacgg
     2761 atctccgagg cctcggaccc gtcgggccgc gtcggacgtg tcagtcctgc tcctcggcca
     2821 cgaagtgcac gcagttgccg gccgggtcgc gcagggcgaa ctcccgcccc cacggctgct
     2881 cgccgatctc ggtcatggcc ggcccggagg cgtcccggaa gttcgtggac acgacctccg
     2941 accactcggc gtacagctcg tccaggccgc gcacccacac ccaggccagg gtgttgtccg
     3001 gcaccacctg gtcctggacc gcgctgatga acagggtcac gtcgtcccgg accacaccgg
     3061 cgaagtcgtc ctccacgaag tcccgggaga acccgagccg gtcggtccag aactcgaccg
     3121 ctccggcgac gtcgcgcgcg gtgagcaccg gaacggcact ggtcaacttg gccatggttt
     3181 agttcctcac cttgtcgtat tatactatgc cgatatacta tgccgatgat taattgtcaa
     3241 caccgccctt agattagatt gctatgcttt ctttctaatg agcaagaagt aaaaaaagtt
     3301 gtaatagaac aagaaaaatg aaactgaaac ttgagaaatt gaagaccgtt tattaactta
     3361 aatatcaatg ggaggtcatc gaaagagaaa aaaatcaaaa aaaaaaattt tcaagaaaaa
     3421 gaaacgtgat aaaaattttt attgcctttt tcgacgaaga aaaagaaacg aggcggtctc
     3481 ttttttcttt tccaaacctt tagtacgggt aattaacgac accctagagg aagaaagagg
     3541 ggaaatttag tatgctgtgc ttgggtgttt tgaagtggta cggcgatgcg cggagtccga
     3601 gaaaatctgg aagagtaaaa aaggagtaga aacattttga agctatggtg tgtgggggat
     3661 ccgcacaaac gaaggtctca cttaatcttc tgtactctga agaggagtgg gaaataccaa
     3721 gaaaaacatc aaactcgaat gattttccca aacccctacc acaagatatt catcagctgc
     3781 gagataggct gatcaggagc aagctcgtac gagaagaaac aaaatgacaa aaaaaatcct
     3841 atactatata ggttacaaat aaaaaagtat caaaaatgaa gcctgcatct ctcaggcaaa
     3901 tggcattctg acatcctctt gattagaatc tagcaagacc ggtcttctcg taagtgccca
     3961 acttgaactg aggaacagtc atgtctaagg ctacaaactc aatgatgatg atgatgatgg
     4021 tcgacggcgc tattcagatc ctcttctgag atgagttttt gttctagaaa gctggcggcc
     4081 gcactttcac cagcctgctt ttcctgttca atcgtttcct tagatggcaa tggattcttc
     4141 tcttgggtct cagttttttt caatttagat ttatcaaact tctctatctc agccatgtcg
     4201 ggcttatcag aggcagaacc tccacttgat cctccagatc ctcctctaga agcgccctga
     4261 aacaatactt caagggatcc accactacct ccacttccac caaagcactt acggtggacg
     4321 atacttggtc ctgattcatc gtattcctgt ttagagatcc acatttgttg gaatgtacta
     4381 agagaggcca atatagaacc gcctatccaa acagaatact tacgctcagg tggtgcgatg
     4441 attttaattt tcatagtaga tggagccaag gccgtgattt ccttttgcat tctatctgca
     4501 atacctgggt acatcgtagt tcctccactc aggaccgtat ttgcgtacaa gtcctttcta
     4561 atagctgcag cacacttcat gatagaattg taagtagtct cgtggattcc acatgcctcc
     4621 attcccaaaa atg
//
