LOCUS       sequence-350934-        6960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Stargazin-mEGFP-iLID in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6960)
  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 6960)
  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-Stargazin-mEGFP-iLID
FEATURES             Location/Qualifiers
     source          1..6960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          667..1687
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1607..1629
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1695..1714
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(2765..2767,2768..2770,2771..3478)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(2810..2831)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3071..3090)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3418..3439
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3512..3949
                     /codon_start=1
                     /product="improved light-inducible dimer when used with 
                     SspB nano or SspB micro (Guntas et al., 2015)"
                     /label=iLID
                     /translation="LATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNVFHLQPMRDYKGDVQ
                     YFIGVQLDGTERLHGAAEREAVCLIKKTAFQIAEAANDENYF"
     primer_bind     complement(3982..4001)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4047..4102
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4101..4120)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4463..4479)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4463..4479)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4476..4498)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4487..4503
                     /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(4511..4517,4518..4535,4536..4541))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4556..4577
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4636..4831
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4682..4817
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4837..4971
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4874..4893)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4928..4947
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5039..5056)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5210..5798)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5290..5309)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5969..6760,6761..6829))
                     /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     6592..6611
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6830..6934)
                     /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 gggggggggn sgcgcgccrg gsggggsggg gsggggsgrg
      541 gggsggggsg gggsgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga
      601 aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg
      661 cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc
      721 cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct
      781 tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg
      841 cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt
      901 gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt
      961 gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg
     1021 gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg
     1081 ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc
     1141 ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg
     1201 gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc
     1261 ggggcggggc cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc
     1321 gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag
     1381 agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc
     1441 cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc
     1501 ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc
     1561 tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg
     1621 cgtgtgaccg gcggctctag ctagagcctc tgctaaccat gttcatgcct tcttcttttt
     1681 cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt
     1741 caccatgggg ctgtttgatc gaggtgttca aatgctttta accaccgttg gtgctttcgc
     1801 tgccttcagc ttgatgacca tcgctgtggg aaccgactat tggctgtact ccagaggggt
     1861 ttgcaagacc aaaagtgtca gtgagaatga aaccagcaag aagaacgagg aagttatgac
     1921 ccattccgga ttatggagaa cctgctgcct cgaagggaac ttcaaaggtc tgtgcaagca
     1981 aatcgaccac tttccggaag acgcggacta cgaagctgac accgcagagt atttcctccg
     2041 ggccgtgagg gcctcgagta tcttcccgat cctgagtgtg atcctgcttt tcatgggtgg
     2101 cctctgcatc gcagccagcg agttctacaa accggtgaac tgcaggaaga gctcgaagaa
     2161 ttcgaaaagc ttgacacgcc acaacatcat cctgagtgct ggcatcttct tcgtgtctgc
     2221 aggtcttagt aatatcatcg ggatcatcgt gtatatatca gccaatgctg gagacccctc
     2281 caagagtgac tccaaaaaga acagctactc ctacggctgg tccttctact tcggggccct
     2341 gtccttcatc atcgccgaga tggtcggggt gctggccgtg cacatgttta tcgaccgcca
     2401 caaacagctg cgggccacgg cccgcgccac cgactacctc caggcctccg ccatcacccg
     2461 catccccagc taccgctacc gctaccagcg ccgcagccgc tccagctcgc gctccaccga
     2521 gccctctcac tccagagacg cctcgcccgt gggcgtgaag ggcttcaaca ccctgccgtc
     2581 cacggagatc tccatgtaca ccctcagtag ggaccccctg aaggctgcca ccacgcccac
     2641 cgccacctac aactcggaca gggataacag ctttctccag gtccacaact gtatccagaa
     2701 ggacagcaag gactctctcc acgccaacac agccaaccgc cggaccacgc ccgtagaatt
     2761 cggcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2821 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     2881 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2941 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     3001 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     3061 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     3121 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     3181 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     3241 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     3301 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     3361 ccactacctg agcacccagt ccaagcttag caaagacccc aacgagaagc gcgatcacat
     3421 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacct
     3481 cgagggtgga tctggaggtt caggtggaag tctggcaacc acactggaac ggatcgagaa
     3541 aaatttcgtg attactgatc cgagactgcc tgacaaccca atcatttttg cgagcgattc
     3601 cttcctgcag ctgacagaat attctcggga agagatcctg gggcgcaatt gccgttttct
     3661 gcagggaccc gagacagacc gtgccactgt tcggaaaatc agagatgcta ttgacaacca
     3721 gactgaagtg accgttcagc tgatcaatta taccaagagc ggcaagaagt tctggaacgt
     3781 gttccacctg cagccgatgc gcgattataa gggcgacgtc cagtacttca ttggcgtgca
     3841 gctggatggc accgaacgtc ttcatggcgc cgctgagcgt gaggcggtct gcctgatcaa
     3901 aaagacagcc tttcagattg ctgaggcagc gaacgacgaa aattactttt aagcggccgc
     3961 ggttctagat aagtcgacgg atctttttcc ctctgccaaa aattatgggg acatcatgaa
     4021 gccccttgag catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg
     4081 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca
     4141 tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa
     4201 caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt
     4261 attccataga aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt
     4321 tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct
     4381 ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacctgca
     4441 gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4501 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4561 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4621 cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4681 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4741 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4801 aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat
     4861 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4921 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt
     4981 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5041 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5101 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5161 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5221 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5281 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5341 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5401 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5461 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5521 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5581 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5641 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5701 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5761 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5821 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5881 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5941 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6001 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6061 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6121 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6181 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6241 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6301 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6361 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6421 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6481 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6541 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6601 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6661 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6721 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6781 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6841 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6901 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
