LOCUS       sequence-326985-        4239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses POLArISact, an orientation probe that specifically binds 
            to F-actin, in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4239)
  AUTHORS   Sugizaki A, Sato K, Chiba K, Saito K, Kawagishi M, Tomabechi Y, 
            Mehta SB, Ishii H, Sakai N, Shirouzu M, Tani T, Terada S
  TITLE     POLArIS, a versatile probe for molecular orientation, revealed actin
            filaments associated with microtubule asters in early embryos.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Mar 16;118(11). pii: 2019071118. doi:
            10.1073/pnas.2019071118.
  PUBMED    33674463
REFERENCE   2  (bases 1 to 4239)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2021-03-16"; volume: "118(11). pii: 
            2019071118. doi"; pages: " 10.1073/pnas.2019071118"
COMMENT     Sequence Label: POLArISact
FEATURES             Location/Qualifiers
     source          1..4239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        127..506
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        507..710
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     661..681
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     755..774
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        755..773
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      787..796
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1411..1458
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             1834..1860
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1907..1925)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1908..1925)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1945..1962)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1951..2175
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2376..2964)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2456..2475)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3135..3926,3927..3995))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3758..3777
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3996..4100)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4175..4194)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
       61 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      121 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      181 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      241 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      301 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      361 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      421 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      481 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      541 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      601 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      661 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa
      721 ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc
      781 aagcttgcca ccatgggagt cgtaccaata cttgtggagt tggacggaga tgtgaacgga
      841 cacaaatttt cagtccgcgg ggagggggaa ggggatgcta ctattggcaa gctgacgctc
      901 aaattcatct gtaccaccgg aaaactccct gtaccctggc ccacactggt gacaactctg
      961 acttacggcg tgcaatgttt tagccgatac ccagaccaca tgaagaggca cgactttttc
     1021 aaaagcgcaa tgcctgaagg atacgtacag gaaaggacca tttcttttaa agacgacggg
     1081 aagtacaaaa cccgggcagt ggtgaagttt gagggcgata ccctcgtcaa taggatcgaa
     1141 ttgaagggaa ctgacttcaa agaagatggc aacatcctgg gtcacaagct tgagtataac
     1201 tttaactccc acaacgtgta tattacagcc gacaaacaga agaatggaat taaggctaac
     1261 ttcactgtca gacacaatgt cgaagatggc tccgtgcagc tcgccgatca ctatcaacag
     1321 aatactccta tcggggacgg cccagtcctg ctgcccgaca accactacct gagtacccag
     1381 actgttctga gcaaagatcc gaacgagaag cgcgaccaca tggtgctgca tgagtatgtc
     1441 aacgctgcag gaattaccct cggcatggac gagctgtaca agggcggtgg atcctccaaa
     1501 ggagaagagc tgttcgaggc cgctgccaag gcttccctgg agatcgaaga actggctcgg
     1561 ttcgctgttg acgaacacaa caaaaaagaa aacgctcttc tagaattcgt tcgcgttgtt
     1621 aaagctaaag aacagagcag cgtgccccac tggtggtgga ccaccatgta ctacctgacc
     1681 ctggaagcta aagacggtgg taaaaagaaa ctgtacgaag ctaaagtttg ggttaaacgc
     1741 gaccccaaca tgatcttcaa gatcaacttc aaagaactgc aggagttcaa accggttggt
     1801 gacgctggcg gaagcggtgg aggcagtggc ggttacccat acgatgttcc agattacgct
     1861 agctgacaca ctgcggccgc tcgagcatgc atctagaggg ccctattcta tagtgtcacc
     1921 taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     1981 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2041 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2101 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2161 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagt ccggaactcg
     2221 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2281 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2341 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2401 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2461 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2521 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2581 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2641 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2701 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2761 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2821 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2881 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2941 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3001 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3061 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3121 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3181 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3241 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     3301 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3361 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3421 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3481 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3541 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3601 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3661 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3721 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     3781 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3841 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3901 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3961 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4021 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4081 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     4141 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     4201 taagccagta tctgctccct gcttgtgtgt tggaggtcg
//
