LOCUS       sequence-366156-        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: I175L
FEATURES             Location/Qualifiers
     source          1..5978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1033..1053)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(1303..1320)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(1338..2276)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     rep_origin      2358..2946
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2847..2866
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3021..3268)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     3234..3252
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(4595..4969)
                     /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(4988..5035)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(5042..5455)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      complement(5470..5716)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(5796..5813)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(5829..5858)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
ORIGIN
        1 gcctatccaa acagaatact tacgctcagg tggtgcgatg attttaattt tcatagtaga
       61 tggagccaag gccgtgattt ccttttgcat tctatctgca atacctgggt acatcgtagt
      121 tcctccactc aggaccgtat ttgcgtacaa gtcctttcta atagctgcag cacacttcat
      181 gatagaattg taagtagtct cgtggattcc acatgcctcc attcccaaaa atgatggctg
      241 gaacagggcc tcaggacatc gaaatctttc gttgccgatg gtaataacct gaccgtctgg
      301 caattcgtaa gacttctcaa gtgaactact agatgcagct gtagccattt cctgttcaaa
      361 atcaagggca acgtaacaca atttttcctt aatatctctg acaatctcac gctcagcagt
      421 tgtggtaaag gagtaccctc gttctgtaag aattttcata aggtaatcag tcaggtcacg
      481 gccagccaaa tctagacgca acaaagcatg aggtaaagca taaccttcgt atattggtac
      541 agtatgagag accccatctc ctgagtctaa aactattccc gtggttctgc ctgaagcata
      601 caaggacaat acagcttgga tggcgacata catggctgga gtgttgaaag tctcgaacat
      661 tatttgggtc attttttctc tattggcttt tggattaaga ggagcctctg tcaataagac
      721 tggatgctct tcaggtgcca ctctcaattc gttgtagaat gtgtgatgcc agattttctc
      781 catgtcatcc cagttagtaa ctataccatg ctcaattggg tacttcaatg tcaatattcc
      841 tcttttagat tgagcttcgt ctcccacgta agagtctttt tgacccatgc cgaccattac
      901 gccttgatga cggggtcgac ctacaataga agggaaaaca gctctagggg cgtcgtcccc
      961 agcaaaacct gctttacaca tacctgaacc gttatccaca accaaggcgg ctacctcctc
     1021 atcacacata tggccttgaa agtacaagtt ttctgatcca ccacttcctc cggatccgcc
     1081 agatccacct gatcctcctg aaccaccact tcctccagaa ccgccagagc caccggatcc
     1141 accactttca ccggcttgtt tttcctgttc aatagtctcc ttactaggta gagggttctt
     1201 ctcctgggtc tctgtttttt ttaacttgga tttgtcaaac ttctcaatct cagccatgtc
     1261 gggcttgtcg gaggcagatc caccagaacc tccggagccg ccgtgatgat gatgatggtg
     1321 acccatggcg aattcctcgt ttcgaataat tagttgtttt ttgatcttct caagttgtcg
     1381 ttaaaagtcg ttaaaatcaa aagcttgtca attggaacca gtcgcaatta tgaaagtaag
     1441 ctaataatga tgataaaaaa aaaggtttaa gacagggcag cttccttctg tttatatatt
     1501 gctgtcaagt aggggttaga acagttaaat tttgatcatg aacgttaggc tatcagcagt
     1561 attcccacca gaatcttgga agcatacaat gtggagacaa tgcataatca tccaaaaagc
     1621 gggtgtttcc ccatttgcgt ttcggcacag gtgcaccggg gttcagaagc gatagagaga
     1681 ctgcgctaag cattaatgag attatttttg agcattcgtc aatcaatacc aaacaagaca
     1741 aacggtatgc cgacttttgg aagtttcttt ttgaccaact ggccgttagc atttcaacga
     1801 accaaactta gttcatcttg gatgagatca cgcttttgtc atattaggtt ccaagacagc
     1861 gtttaaactg tcagttttgg gccatttggg gaacatgaaa ctatttgacc ccacactcag
     1921 aaagccctca tctggagtga tgttcgggtg taatgcggag cttgttgcat tcggaaataa
     1981 acaaacatga acctcgccag gggggccagg atagacaggc taataaagtc atggtgttag
     2041 tagcctaata gaaggaattg gaatgagcga gctccaatca agcccaataa ctgggctggt
     2101 ttttcgatgg caaaagtggg tgttgaggag aagaggagtg gaggtcctgc gtttgcaacg
     2161 gtctgctgct agtgtatccc ctcctgttgc gtttggcact tatgtgtgag aatggacctg
     2221 tggatgtcgg atggcaaaaa ggtttcattc aacctttcgt ctttggatgt tagatctgat
     2281 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2341 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2401 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2461 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2521 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2581 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2641 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2701 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2761 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2821 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2881 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2941 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3001 cacatgttgg tctccagctt gcaaattaaa gccttcgagc gtcccaaaac cttctcaagc
     3061 aaggttttca gtataatgtt acatgcgtac acgcgtctgt acagaaaaaa aagaaaaatt
     3121 tgaaatataa ataacgttct taatactaac ataactataa aaaaataaat agggacctag
     3181 acttcaggtt gtctaactcc ttccttttcg gttagagcgg atgtgggggg agggcgtgaa
     3241 tgtaagcgtg acataactaa ttacatgata tcgacaaagg aaaaggggga cggatctccg
     3301 aggcctcgtg atcctaccca cgtaatatgg acacaggcct aagcgaggtt cttgttttca
     3361 aattgttccg gactgaggcc gccacaccaa ctgtgccgcc gccaccgatt gtaatcacat
     3421 tcgatataat taaacaccgt tgcccgcatt atttcccggc tgataaagtg ttctccatgg
     3481 atacattcca ctttcagcga atgaaagaag ctttccacgc aggcattatc gtagcagcaa
     3541 ccttttgcgc tcatacttcc acgcagatta tgccgcttca gttgcgcctg ataatctgct
     3601 gaacagtact ggcctccacg gtccgtgtga acgataacgt tccggggcct cttacgccgc
     3661 cacagcgcca tctgcagggc atcgcaggcc agttgcgccg tcatgcgtgg cgacattgac
     3721 cagccaataa cggcacgtga ccacaggtca atgaccactg ccagatacag ccagccttca
     3781 tctgtacgta agtacgtgat gtctcctgcc cacttctggt tcgggccact ggcgtaaaaa
     3841 tcctgctcca acagattttc tgacacaggc aggccgtgtg cgcggtagct gaccgggctg
     3901 aacttccggg aggcctttgc cctcagtccc tgacggcgca ggcttgccgc cacggttttt
     3961 acgttaaagg ggtaaccctg agcacgcagt tcatccgtca ggcgtggggc accgtaacgc
     4021 tgttttgacc gggtaaaagc cgcgaggaca acgctgtcgc agtgttggcg gaactgctga
     4081 cgcgtgctta tccttgtccg ccgctgacac cacgtatacc agccgctgcg ggccacccgg
     4141 agcacgcggc acattgcttt gatgctgaac tcagcctgat gtttttcaat aaagacatac
     4201 ttcatttcag gcgcttcgcg aagtatgtcg cggccttttg gaggatagcc agctcttcat
     4261 cccgttctgc cagctggcgt ttgagacgtg caatctcggt agacatctcc agttcacgtt
     4321 cagaagacgt ctgctgattt tgctgtttac tgcgccagtt gtagagctgt gattcataca
     4381 ggctgagttc acgggctgcg gcagtaacac cgatgcgttc agcaagcttc agggcttcac
     4441 tgcgaaattc aggcgaatgc tgtttacggg gttttttact ggttgatact gtttttgtca
     4501 tgtgagtcac ctctgactga gagtttactc acttagccgc gtgtccacta ttgctgggta
     4561 agatcatcgg acccgtcggg ccgcgtcgga cgtgtcagtc ctgctcctcg gccacgaagt
     4621 gcacgcagtt gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga
     4681 tctcggtcat ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact
     4741 cggcgtacag ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg tccggcacca
     4801 cctggtcctg gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt
     4861 cgtcctccac gaagtcccgg gagaacccga gccggtcggt ccagaactcg accgctccgg
     4921 cgacgtcgcg cgcggtgagc accggaacgg cactggtcaa cttggccatg gtttagttcc
     4981 tcaccttgtc gtattatact atgccgatat actatgccga tgattaattg tcaacaccgc
     5041 ccttagatta gattgctatg ctttctttct aatgagcaag aagtaaaaaa agttgtaata
     5101 gaacaagaaa aatgaaactg aaacttgaga aattgaagac cgtttattaa cttaaatatc
     5161 aatgggaggt catcgaaaga gaaaaaaatc aaaaaaaaaa attttcaaga aaaagaaacg
     5221 tgataaaaat ttttattgcc tttttcgacg aagaaaaaga aacgaggcgg tctctttttt
     5281 cttttccaaa cctttagtac gggtaattaa cgacacccta gaggaagaaa gaggggaaat
     5341 ttagtatgct gtgcttgggt gttttgaagt ggtacggcga tgcgcggagt ccgagaaaat
     5401 ctggaagagt aaaaaaggag tagaaacatt ttgaagctat ggtgtgtggg ggatccgcac
     5461 aaacgaaggt ctcacttaat cttctgtact ctgaagagga gtgggaaata ccaagaaaaa
     5521 catcaaactc gaatgatttt cccaaacccc taccacaaga tattcatcag ctgcgagata
     5581 ggctgatcag gagcaagctc gtacgagaag aaacaaaatg acaaaaaaaa tcctatacta
     5641 tataggttac aaataaaaaa gtatcaaaaa tgaagcctgc atctctcagg caaatggcat
     5701 tctgacatcc tcttgattag aatctagcaa gaccggtctt ctcgtaagtg cccaacttga
     5761 actgaggaac agtcatgtct aaggctacaa actcaatgat gatgatgatg atggtcgacg
     5821 gcgctattca gatcctcttc tgagatgagt ttttgttcta gaaagctggc ggccgcttaa
     5881 aagcacttac ggtggacgat acttggtcct gattcatcgt attcctgttt agagatccac
     5941 atttgttgga atgtactaag agaggccaat atagaacc
//
