LOCUS       sequence-362191-        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: I250L
FEATURES             Location/Qualifiers
     source          1..4633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(909..1847)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     rep_origin      1929..2517
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2418..2437
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2592..2839)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2805..2823
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(2905..3279)
                     /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(3298..3345)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(3352..3765)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      complement(3780..4026)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(4106..4123)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4139..4168)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
ORIGIN
        1 caggaccgta tttgcgtaca agtcctttct aatagctgca gcacacttca tgatagaatt
       61 gtaagtagtc tcgtggattc cacatgcctc cattcccaaa aatgatggct ggaacagggc
      121 ctcaggacat cgaaatcttt cgttgcccaa ggtaataacc tgaccgtctg gcaattcgta
      181 agacttctca agtgaactac tagatgcagc tgtagccatt tcctgttcaa aatcaagggc
      241 aacgtaacac aatttttcct taatatctct gacaatctca cgctcagcag ttgtggtaaa
      301 ggagtaccct cgttctgtaa gaattttcat aaggtaatca gtcaggtcac ggccagccaa
      361 atctagacgc aaaatagcat gaggtaaagc ataaccttcg tatattggta cagtatgaga
      421 gaccccatct cctgagtcta aaactattcc cgtggttctg cctgaagcat acaaggacaa
      481 tacagcttgg atggcgacat acatggctgg agtgttgaaa gtctcgaaca ttatttgggt
      541 cattttttct ctattggctt ttggattaag aggagcctct gtcaataaga ctggatgctc
      601 ttcaggtgcc actctcaatt cgttgtagaa tgtgtgatgc cagattttct ccatgtcatc
      661 ccagttagta actataccat gctcaattgg gtacttcaat gtcaatattc ctcttttaga
      721 ttgagcttcg tctcccacgt aagagtcttt ttgacccatg ccgaccatta cgccttgatg
      781 acggggtcga cctacaatag aagggaaaac agctctaggg gcgtcgtccc cagcaaaacc
      841 tgctttacac atacctgaac cgttatccac aaccaaggcg gctacctcct catcacacat
      901 gaattcctcg tttcgaataa ttagttgttt tttgatcttc tcaagttgtc gttaaaagtc
      961 gttaaaatca aaagcttgtc aattggaacc agtcgcaatt atgaaagtaa gctaataatg
     1021 atgataaaaa aaaaggttta agacagggca gcttccttct gtttatatat tgctgtcaag
     1081 taggggttag aacagttaaa ttttgatcat gaacgttagg ctatcagcag tattcccacc
     1141 agaatcttgg aagcatacaa tgtggagaca atgcataatc atccaaaaag cgggtgtttc
     1201 cccatttgcg tttcggcaca ggtgcaccgg ggttcagaag cgatagagag actgcgctaa
     1261 gcattaatga gattattttt gagcattcgt caatcaatac caaacaagac aaacggtatg
     1321 ccgacttttg gaagtttctt tttgaccaac tggccgttag catttcaacg aaccaaactt
     1381 agttcatctt ggatgagatc acgcttttgt catattaggt tccaagacag cgtttaaact
     1441 gtcagttttg ggccatttgg ggaacatgaa actatttgac cccacactca gaaagccctc
     1501 atctggagtg atgttcgggt gtaatgcgga gcttgttgca ttcggaaata aacaaacatg
     1561 aacctcgcca ggggggccag gatagacagg ctaataaagt catggtgtta gtagcctaat
     1621 agaaggaatt ggaatgagcg agctccaatc aagcccaata actgggctgg tttttcgatg
     1681 gcaaaagtgg gtgttgagga gaagaggagt ggaggtcctg cgtttgcaac ggtctgctgc
     1741 tagtgtatcc cctcctgttg cgtttggcac ttatgtgtga gaatggacct gtggatgtcg
     1801 gatggcaaaa aggtttcatt caacctttcg tctttggatg ttagatctga tctcatgacc
     1861 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1921 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1981 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2041 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     2101 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2161 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2221 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2281 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2341 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2401 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2461 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2521 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttg
     2581 gtctccagct tgcaaattaa agccttcgag cgtcccaaaa ccttctcaag caaggttttc
     2641 agtataatgt tacatgcgta cacgcgtctg tacagaaaaa aaagaaaaat ttgaaatata
     2701 aataacgttc ttaatactaa cataactata aaaaaataaa tagggaccta gacttcaggt
     2761 tgtctaactc cttccttttc ggttagagcg gatgtggggg gagggcgtga atgtaagcgt
     2821 gacataacta attacatgat atcgacaaag gaaaaggggg acggatctcc gaggcctcgg
     2881 acccgtcggg ccgcgtcgga cgtgtcagtc ctgctcctcg gccacgaagt gcacgcagtt
     2941 gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat
     3001 ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag
     3061 ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg
     3121 gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac
     3181 gaagtcccgg gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg
     3241 cgcggtgagc accggaacgg cactggtcaa cttggccatg gtttagttcc tcaccttgtc
     3301 gtattatact atgccgatat actatgccga tgattaattg tcaacaccgc ccttagatta
     3361 gattgctatg ctttctttct aatgagcaag aagtaaaaaa agttgtaata gaacaagaaa
     3421 aatgaaactg aaacttgaga aattgaagac cgtttattaa cttaaatatc aatgggaggt
     3481 catcgaaaga gaaaaaaatc aaaaaaaaaa attttcaaga aaaagaaacg tgataaaaat
     3541 ttttattgcc tttttcgacg aagaaaaaga aacgaggcgg tctctttttt cttttccaaa
     3601 cctttagtac gggtaattaa cgacacccta gaggaagaaa gaggggaaat ttagtatgct
     3661 gtgcttgggt gttttgaagt ggtacggcga tgcgcggagt ccgagaaaat ctggaagagt
     3721 aaaaaaggag tagaaacatt ttgaagctat ggtgtgtggg ggatccgcac aaacgaaggt
     3781 ctcacttaat cttctgtact ctgaagagga gtgggaaata ccaagaaaaa catcaaactc
     3841 gaatgatttt cccaaacccc taccacaaga tattcatcag ctgcgagata ggctgatcag
     3901 gagcaagctc gtacgagaag aaacaaaatg acaaaaaaaa tcctatacta tataggttac
     3961 aaataaaaaa gtatcaaaaa tgaagcctgc atctctcagg caaatggcat tctgacatcc
     4021 tcttgattag aatctagcaa gaccggtctt ctcgtaagtg cccaacttga actgaggaac
     4081 agtcatgtct aaggctacaa actcaatgat gatgatgatg atggtcgacg gcgctattca
     4141 gatcctcttc tgagatgagt ttttgttcta gaaagctggc ggccgcactt tcaccagcct
     4201 gcttttcctg ttcaatcgtt tccttagatg gcaatggatt cttctcttgg gtctcagttt
     4261 ttttcaattt agatttatca aacttctcta tctcagccat gtcgggctta tcagaggcag
     4321 aacctccact tgatcctcca gatcctcctc tagaagcgcc ctgaaacaat acttcaaggg
     4381 atccaccact acctccactt ccaccaaagc acttacggtg gacgatactt ggtcctgatt
     4441 catcgtattc ctgtttagag atccacattt gttggaatgt actaagagag gccaatatag
     4501 aaccgcctat ccaaacagaa tacttacgct caggtggtgc gatgatttta attttcatag
     4561 tagatggagc caaggccgtg atttcctttt gcattctatc tgcaatacct gggtacatcg
     4621 tagttcctcc act
//
