LOCUS       sequence-326987-        4329 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flexible linker control for POLArISact expression in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4329)
  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 4329)
  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: cpGFP-Adhiron_flex
FEATURES             Location/Qualifiers
     source          1..4329
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        137..516
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        517..720
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     671..691
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     765..784
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        765..783
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      797..806
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1421..1468
                     /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"
     regulatory      1589..1598
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1934..1960
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(2007..2025)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2008..2025)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2045..2062)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2051..2275
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2476..3064)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2556..2575)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3235..4026,4027..4095))
                     /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     3858..3877
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4096..4200)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4275..4294)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
       61 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
      121 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
      181 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      241 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      301 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      361 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      421 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      481 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      541 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      601 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      661 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      721 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat
      781 agggagaccc aagcttgcca ccatgggagt cgtaccaata cttgtggagt tggacggaga
      841 tgtgaacgga cacaaatttt cagtccgcgg ggagggggaa ggggatgcta ctattggcaa
      901 gctgacgctc aaattcatct gtaccaccgg aaaactccct gtaccctggc ccacactggt
      961 gacaactctg acttacggcg tgcaatgttt tagccgatac ccagaccaca tgaagaggca
     1021 cgactttttc aaaagcgcaa tgcctgaagg atacgtacag gaaaggacca tttcttttaa
     1081 agacgacggg aagtacaaaa cccgggcagt ggtgaagttt gagggcgata ccctcgtcaa
     1141 taggatcgaa ttgaagggaa ctgacttcaa agaagatggc aacatcctgg gtcacaagct
     1201 tgagtataac tttaactccc acaacgtgta tattacagcc gacaaacaga agaatggaat
     1261 taaggctaac ttcactgtca gacacaatgt cgaagatggc tccgtgcagc tcgccgatca
     1321 ctatcaacag aatactccta tcggggacgg cccagtcctg ctgcccgaca accactacct
     1381 gagtacccag actgttctga gcaaagatcc gaacgagaag cgcgaccaca tggtgctgca
     1441 tgagtatgtc aacgctgcag gaattaccct cggcatggac gagctgtaca agggcggtgg
     1501 atcctccaaa ggagaagagc tgttcactga agcggctgca aagggaggat ccggaggcag
     1561 cgccggttca gcagccggtt ctggcggtgc caccatggct accggtgttc gcgcagttcc
     1621 gggtaacgaa aactccctgg aaatcgaaga actggctcgg ttcgctgttg acgaacacaa
     1681 caaaaaagaa aacgctcttc tagaattcgt tcgcgttgtt aaagctaaag aacagagcag
     1741 cgtgccccac tggtggtgga ccaccatgta ctacctgacc ctggaagcta aagacggtgg
     1801 taaaaagaaa ctgtacgaag ctaaagtttg ggttaaacgc gaccccaaca tgatcttcaa
     1861 gatcaacttc aaagaactgc aggagttcaa accggttggt gacgctggcg gaagcggtgg
     1921 aggcagtggc ggttacccat acgatgttcc agattacgct agctgacaca ctgcggccgc
     1981 tcgagcatgc atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga
     2041 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2101 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2161 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2221 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2281 gaggcggaaa gaaccagctg gggctctagt ccggaactcg ctgcgctcgg tcgttcggct
     2341 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2401 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2461 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2521 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2581 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2641 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2701 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2761 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2821 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2881 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2941 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3001 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3061 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3121 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3181 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3241 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3301 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3361 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3421 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3481 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3541 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3601 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3661 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3721 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3781 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3841 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3901 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3961 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4021 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4081 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4141 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4201 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     4261 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     4321 gcttgtgtg
//
