LOCUS       sequence-214076-        4798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses arginylated human beta-actin fused with thymosin beta and 
            a His tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4798)
  AUTHORS   Hatano T, Alioto S, Roscioli E, Palani S, Clarke ST, Kamnev A, 
            Hernandez-Fernaud JR, Sivashanmugam L, Chapa-Y-Lazo B, Jones AME, 
            Robinson RC, Sampath K, Mishima M, McAinsh AD, Goode BL, 
            Balasubramanian MK
  TITLE     Rapid production of pure recombinant actin isoforms in Pichia 
            pastoris.
  JOURNAL   J Cell Sci. 2018 Mar 13. pii: jcs.213827. doi: 10.1242/jcs.213827.
  PUBMED    29535210
REFERENCE   2  (bases 1 to 4798)
  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"; journalName: "J Cell Sci.
            2018 Mar 13. pii: jcs.213827. doi: 10.1242/jcs.213827."
COMMENT     Sequence Label: pPICZc-Ub-R-HsACTB*-thymosinB-8His
FEATURES             Location/Qualifiers
     source          1..4798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..940
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             955..1182
                     /codon_start=1
                     /product="Saccharomyces cerevisiae ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVESSDTIDNVKSKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             2482..2502
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2506..2529
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      2621..2867
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2882..3295
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3302..3349
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3368..3742
                     /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      3808..4055
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3824..3842)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(4130..4718)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4210..4229)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag
       61 gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt
      121 tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc
      181 agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta
      241 acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta
      301 tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg
      361 agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct
      421 gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg
      481 ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt
      541 cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct
      601 ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct
      661 ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact
      721 gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat
      781 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt
      841 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga
      901 caacttgaga agatcaaaaa acaactaatt attcgaaacg aggaattcgc caccatgcag
      961 attttcgtca agactttgac cggtaaaacc ataacattgg aagttgaatc ttccgatacc
     1021 atcgacaacg ttaagtcgaa aattcaagac aaggaaggta tccctccaga tcaacaaaga
     1081 ttgatctttg ccggtaagca gctagaagac ggtagaacgc tgtctgatta caacattcag
     1141 aaggagtcca ccttacatct tgtgctaagg ctaagaggtg gtagagatga tatcgccgcg
     1201 ctcgtcgtcg acaacggctc cggcatgtgc aaggccggct tcgcgggcga cgatgccccc
     1261 cgggccgtct tcccctccat cgtggggcgc cccaggcacc agggcgtgat ggtgggcatg
     1321 ggtcagaagg attcctatgt gggcgacgag gcccagagca agagaggcat cctcaccctg
     1381 aagtacccca tcgagcacgg catcgtcacc aactgggacg acatggagaa aatctggcac
     1441 cacaccttct acaatgagct gcgtgtggct cccgaggagc accccgtgct gctgaccgag
     1501 gcccccctga accccaaggc caaccgcgag aagatgaccc agatcatgtt tgagaccttc
     1561 aacaccccag ccatgtacgt tgctatccag gctgtgctat ccctgtacgc ctctggccgt
     1621 accactggca tcgtgatgga ctccggtgac ggggtcaccc acactgtgcc catctacgag
     1681 gggtatgccc tcccccatgc catcctgcgt ctggacctgg ctggccggga cctgactgac
     1741 tacctcatga agatcctcac cgagcgcggc tacagcttca ccaccacggc cgagcgggaa
     1801 atcgtgcgtg acattaagga gaagctgtgc tacgtcgccc tggacttcga gcaagagatg
     1861 gccacggctg cttccagctc ctccctggag aagagctacg agctgcctga cggccaggtc
     1921 atcaccattg gcaatgagcg gttccgctgc cctgaggcac tcttccagcc ttccttcctg
     1981 ggcatggagt cctgtggcat ccacgaaact accttcaact ccatcatgaa gtgtgacgtg
     2041 gacatccgca aagacctgta cgccaacaca gtgctgtctg gcggcaccac catgtaccct
     2101 ggcattgccg acaggatgca gaaggagatc actgccctgg cacccagcac aatgaagatc
     2161 aagatcattg ctcctcctga gcgcaagtac tccgtgtgga tcggcggctc aatcctggcc
     2221 tcgctgtcca ccttccagca gatgtggatc agcaagcagg agtatgacga gtccggcccc
     2281 tccatcgtcc accgcaaatg cttcgctagc cgtggcggca gcggtgggag ctctggcggt
     2341 agcgcttccg ataagccaga tatggcggaa atcgaaaagt tcgataaatc taaactgaag
     2401 aaaactgaga ctcaagaaaa gaacccgctg ccgagcaagg aaaccatcga gcaggagaag
     2461 caggcgggtg aatccggtac cgaaaattta tattttcaag ggccccacca ccaccaccac
     2521 caccaccacg aagaataact taagtcaatt cgccttagac atgactgttc ctcagttcaa
     2581 gttgggcact tacgagaaga ccggtcttgc tagattctaa tcaagaggat gtcagaatgc
     2641 catttgcctg agagatgcag gcttcatttt tgatactttt ttatttgtaa cctatatagt
     2701 ataggatttt ttttgtcatt ttgtttcttc tcgtacgagc ttgctcctga tcagcctatc
     2761 tcgcagctga tgaatatctt gtggtagggg tttgggaaaa tcattcgagt ttgatgtttt
     2821 tcttggtatt tcccactcct cttcagagta cagaagatta agtgagacct tcgtttgtgc
     2881 ggatccccca cacaccatag cttcaaaatg tttctactcc ttttttactc ttccagattt
     2941 tctcggactc cgcgcatcgc cgtaccactt caaaacaccc aagcacagca tactaaattt
     3001 cccctctttc ttcctctagg gtgtcgttaa ttacccgtac taaaggtttg gaaaagaaaa
     3061 aagagaccgc ctcgtttctt tttcttcgtc gaaaaaggca ataaaaattt ttatcacgtt
     3121 tctttttctt gaaaattttt ttttttgatt tttttctctt tcgatgacct cccattgata
     3181 tttaagttaa taaacggtct tcaatttctc aagtttcagt ttcatttttc ttgttctatt
     3241 acaacttttt ttacttcttg ctcattagaa agaaagcata gcaatctaat ctaagggcgg
     3301 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     3361 ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg
     3421 agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt
     3481 cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt
     3541 gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg
     3601 gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg
     3661 cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt
     3721 cgtggccgag gagcaggact gacacgtccg acgcggcccg acgggtccga ggcctcggag
     3781 atccgtcccc cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac
     3841 gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     3901 tccctattta tttttttata gttatgttag tattaagaac gttatttata tttcaaattt
     3961 ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg
     4021 agaaggtttt gggacgctcg aaggctttaa tttgcaagct ggagaccaac atgtgagcaa
     4081 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4141 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4201 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4261 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4321 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4381 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4441 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4501 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4561 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4621 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4681 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4741 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagatc
//
