LOCUS       sequence-350937-        7493 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CRY2-mCherry-MYPT (1-169 aa) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7493)
  AUTHORS   Yamamoto K, Miura H, Ishida M, Mii Y, Kinoshita N, Takada S, Ueno N,
            Sawai S, Kondo Y, Aoki K
  TITLE     Optogenetic relaxation of actomyosin contractility uncovers 
            mechanistic roles of cortical tension during cytokinesis.
  JOURNAL   Nat Commun. 2021 Dec 8;12(1):7145. doi: 10.1038/s41467-021-27458-3.
  PUBMED    34880255
REFERENCE   2  (bases 1 to 7493)
  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: "Nat 
            Commun."; date: "2021-12-8"; volume: "12(1):7145. doi"; pages: " 
            10.1038/s41467-021-27458-3"
COMMENT     Sequence Label: pCAGGS-CRY2-mCherry-MYPT169
FEATURES             Location/Qualifiers
     source          1..7493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        8..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          661..1681
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1601..1623
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1689..1708
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1739..3232
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     CDS             3239..3943
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTLGMDELY"
     primer_bind     complement(3384..3402)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3652..3671
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4472..4498
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(4514..4533)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4579..4634
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4633..4652)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4995..5011)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4995..5011)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5008..5030)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5019..5035
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5043..5049,5050..5067,5068..5073))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5088..5109
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5168..5363
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5214..5349
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5369..5503
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5406..5425)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5460..5479
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5571..5588)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5742..6330)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5822..5841)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6501..7292,7293..7361))
                     /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     7124..7143
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7362..7466)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgatcgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc
      421 cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt
      481 gcagcgatgg gggcgggggg gggggggggg gggggggggg cggggcgggg cgaggggcgg
      541 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
      721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
      901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
      961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
     1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
     1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
     1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
     1261 gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
     1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
     1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
     1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     1621 accggcggct ctagctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca
     1681 gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattcaccat
     1741 gaagatggac aaaaagacta tagtttggtt tagaagagac ctaaggattg aggataatcc
     1801 tgcattagca gcagctgctc acgaaggatc tgtttttcct gtcttcattt ggtgtcctga
     1861 agaagaagga cagttttatc ctggaagagc ttcaagatgg tggatgaaac aatcacttgc
     1921 tcacttatct caatccttga aggctcttgg atctgacctc actttaatca aaacccacaa
     1981 cacgatttca gcgatcttgg attgtatccg cgttaccggt gctacaaaag tcgtctttaa
     2041 ccacctctat gatcctgttt cgttagttcg ggaccatacc gtaaaggaga agctggtgga
     2101 acgtgggatc tctgtgcaaa gctacaatgg agatctattg tatgaaccgt gggagatata
     2161 ctgcgaaaag ggcaaacctt ttacgagttt caattcttac tggaagaaat gcttagatat
     2221 gtcgattgaa tccgttatgc ttcctcctcc ttggcggttg atgccaataa ctgcagcggc
     2281 tgaagcgatt tgggcgtgtt cgattgaaga actagggctg gagaatgagg ccgagaaacc
     2341 gagcaatgcg ttgttaacta gagcttggtc tccaggatgg agcaatgctg ataagttact
     2401 aaatgagttc atcgagaagc agttgataga ttatgcaaag aacagcaaga aagttgttgg
     2461 gaactctact tcactacttt ctccgtatct ccatttcggg gaaataagcg tcagacacgt
     2521 tttccagtgt gcccggatga aacaaattat atgggcaaga gataagaaca gtgaaggaga
     2581 agaaagtgca gatctttttc ttaggggaat cggtttaaga gagtattctc ggtatatatg
     2641 tttcaacttc ccgtttactc acgagcaatc gttgttgagt catcttcggt ttttcccttg
     2701 ggatgctgat gttgataagt tcaaggcctg gagacaaggc aggaccggtt atccgttggt
     2761 ggatgccgga atgagagagc tttgggctac cggatggatg cataacagaa taagagtgat
     2821 tgtttcaagc tttgctgtga agtttcttct ccttccatgg aaatggggaa tgaagtattt
     2881 ctgggataca cttttggatg ctgatttgga atgtgacatc cttggctggc agtatatctc
     2941 tgggagtatc cccgatggcc acgagcttga tcgcttggac aatcccgcgt tacaaggcgc
     3001 caaatatgac ccagaaggtg agtacataag gcaatggctt cccgagcttg cgagattgcc
     3061 aactgaatgg atccatcatc catgggacgc tcctttaacc gtactcaaag cttctggtgt
     3121 ggaactcgga acaaactatg cgaaacccat tgtagacatc gacacagctc gtgagctact
     3181 agctaaagct atttcaagaa cccgtgaagc acagatcatg atcggagcag cactcgagat
     3241 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     3301 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     3361 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     3421 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     3481 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     3541 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     3601 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     3661 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     3721 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     3781 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     3841 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     3901 cgccgagggc cgccactcca ctctcggcat ggacgagctg tacggcggcc gcatgaagat
     3961 ggcggacgcg aagcagaagc ggaacgagca gctgaaacgc tggatcggct ccgagacgga
     4021 cctcgagcct ccggtggtga agcgccagaa gaccaaggtg aagttcgacg atggcgccgt
     4081 cttcctggct gcttgctcca gcggcgacac ggacgaggtc ctcaagctgc tgcaccgcgg
     4141 cgccgacatc aattacgcca atgtggacgg actcactgcc ctgcaccagg cttgcattga
     4201 tgacaatgtt gatatggtga agtttctggt agaaaatgga gcaaatatta atcaacctga
     4261 taatgaaggc tggataccac tacatgcagc agcttcctgt ggatatcttg atattgcaga
     4321 gtttttgatt ggtcaaggag cacatgtagg ggctgtcaac agtgaaggag atacaccttt
     4381 agatattgcg gaggaggagg caatggaaga gctacttcaa aatgaagtta atcggcaagg
     4441 ggttgatata gaagcagctt ctagactgca gctgcctcct ctggaacgcc tgactctgga
     4501 ttaagtcgac ggatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg
     4561 agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     4621 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg
     4681 agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt
     4741 ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata
     4801 gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt
     4861 taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac
     4921 tactcccagt catagctgtc cctcttctct tatggagatc cctcgacctg cagcccaagc
     4981 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5041 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5101 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5161 cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5221 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5281 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     5341 ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata atggttacaa
     5401 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5461 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca ttaatgaatc
     5521 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5581 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5641 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5701 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5761 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5821 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5881 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5941 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6001 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6061 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6121 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6181 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6241 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6301 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6361 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6421 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6481 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6541 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6601 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6661 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6721 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6781 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6841 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6901 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6961 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7021 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7081 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7141 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7201 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7261 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7321 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7381 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7441 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgg
//
