LOCUS       sequence-350940-        6674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PP1c-miRFP703 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6674)
  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 6674)
  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-PPP1CB-miRFP703
FEATURES             Location/Qualifiers
     source          1..6674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..665
                     /label=chicken beta-actin promoter
     intron          666..1686
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1606..1628
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1694..1713
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2733..3677
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 703 nm (Shcherbakova et al., 2016)"
                     /label=miRFP703
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSLVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPIHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     primer_bind     complement(3696..3715)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3761..3816
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3815..3834)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4177..4193)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4177..4193)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4190..4212)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4201..4217
                     /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(4225..4231,4232..4249,4250..4255))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4270..4291
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4350..4545
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4396..4531
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4551..4685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4588..4607)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4642..4661
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4753..4770)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4924..5512)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5004..5023)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5683..6474,6475..6543))
                     /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     6306..6325
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6544..6648)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgatcga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
      421 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      481 tgcagcgatg ggggcggggg gggggggggs gcgcgccrgg sggggsgggg sggggsgrgg
      541 ggsggggsgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
      601 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
      661 gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc
      721 gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt
      781 ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc
      841 gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg
      901 cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg
      961 agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg
     1021 ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg
     1081 gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc
     1141 tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg
     1201 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
     1261 gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg
     1321 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
     1381 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
     1441 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
     1501 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
     1561 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     1621 gtgtgaccgg cggctctagc tagagcctct gctaaccatg ttcatgcctt cttctttttc
     1681 ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg caagaattcg
     1741 ccatggcgga cggggagctg aacgtggaca gcctcatcac ccggctgctg gaggtacgag
     1801 gatgtcgtcc aggaaagatt gtgcagatga ctgaagcaga agttcgaggc ttatgtatca
     1861 agtctcggga gatctttctc agccagccta ttcttttgga attggaagca ccgctgaaaa
     1921 tttgtggaga tattcatgga cagtatacag atttactgag attatttgaa tatggaggtt
     1981 tcccaccaga agccaactat cttttcttag gagattatgt ggacagagga aagcagtctt
     2041 tggaaaccat ttgtttgcta ttggcttata aaatcaaata tccagagaac ttctttctct
     2101 taagaggaaa ccatgagtgt gctagcatca atcgcattta tggattctat gatgaatgca
     2161 aacgaagatt taatattaaa ttgtggaaga ccttcactga ttgttttaac tgtctgccta
     2221 tagcagccat tgtggatgag aagatcttct gttgtcatgg aggattgtca ccagacctgc
     2281 aatctatgga gcagattcgg agaattatga gacctactga tgtccctgat acaggtttgc
     2341 tctgtgattt gctatggtct gatccagata aggatgtgca aggctgggga gaaaatgatc
     2401 gtggtgtttc ctttactttt ggagctgatg tagtcagtaa atttctgaat cgtcatgatt
     2461 tagatttgat ttgtcgagct catcaggtgg tggaagatgg atatgaattt tttgctaaac
     2521 gacagttggt aaccttattt tcagccccaa attactgtgg cgagtttgat aatgctggtg
     2581 gaatgatgag tgtggatgaa actttgatgt gttcatttca gatattgaaa ccatctgaaa
     2641 agaaagctaa ataccagtat ggtggactga attctggacg tcctgtcact ccacctcgaa
     2701 cagctaatcc gccgaagaaa aggggcggcc gcatggtagc aggtcatgcc tctggcagcc
     2761 ccgcattcgg gaccgcctct cattcgaatt gcgaacatga agagatccac ctcgccggct
     2821 cgatccagcc gcatggcgcg cttctggtcg tcagcgaaca tgatcatcgc gtcatccagg
     2881 ccagcgccaa cgccgcggaa tttctgaatc tcggaagcgt actcggcgtt ccgctcgccg
     2941 agatcgacgg cgatctgttg atcaagatcc tgccgcatct cgatcccacc gccgaaggca
     3001 tgccggtcgc ggtgcgctgc cggatcggca atccctctac ggagtactgc ggtctgatgc
     3061 atcggcctcc ggaaggcggg ctgatcatcg aactcgaacg tgccggcccg tcgatcgatc
     3121 tgtcaggcac gctggcgccg gcgctggagc ggatccgcac ggcgggttca ctgcgcgcgc
     3181 tgtgcgatga caccgtgctg ctgtttcagc agtgcaccgg ctacgaccgg gtgatggtgt
     3241 atcgtttcga tgagcaaggc cacggcctgg tattctccga gtgccatgtg cctgggctcg
     3301 aatcctattt cggcaaccgc tatccgtcgt cgctggtccc gcagatggcg cggcagctgt
     3361 acgtgcggca gcgcgtccgc gtgctggtcg acgtcaccta tcagccggtg ccgctggagc
     3421 cgcggctgtc gccgctgacc gggcgcgatc tcgacatgtc gggctgcttc ctgcgctcga
     3481 tgtcgccgat ccatctgcag ttcctgaagg acatgggcgt gcgcgccacc ctggcggtgt
     3541 cgctggtggt cggcggcaag ctgtggggcc tggttgtctg tcaccattat ctgccgcgct
     3601 tcatccgttt cgagctgcgg gcgatctgca aacggctcgc cgaaaggatc gcgacgcgga
     3661 tcaccgcgct tgagagctct agataagtcg acggatcttt ttccctctgc caaaaattat
     3721 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     3781 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     3841 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     3901 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     3961 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     4021 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     4081 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     4141 tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4201 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4261 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4321 gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg
     4381 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4441 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4501 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta
     4561 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4621 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     4681 ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4741 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4801 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4861 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4921 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4981 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5041 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5101 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5161 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5221 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5281 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5341 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5401 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5461 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5521 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5581 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5641 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5701 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5761 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5821 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5881 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5941 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6001 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6061 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6121 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6181 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6241 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6301 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6361 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6421 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6481 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6541 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6601 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6661 aaaagtgcca cctg
//
