LOCUS       sequence-485113-        6870 bp    DNA     circular SYN 02-SEP-2025
DEFINITION  Expresses a localizer of Opto-p53 in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6870)
  AUTHORS   Tsuruoka T, Goto Y, Aoki K
  TITLE     Opto-p53: A Light-Controllable Activation of p53 Signaling Pathway.
  JOURNAL   Cell Struct Funct. 2025 Jun 12. doi: 10.1247/csf.25017.
  PUBMED    40500178
REFERENCE   2  (bases 1 to 6870)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Struct
            Funct. 2025 Jun 12. doi: 10.1247/csf.25017."
COMMENT     Sequence Label: pCAGGS-CIBN-p53CT(98-393)-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..6870
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     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             1744..2253
                     /codon_start=1
                     /product="N-terminal portion of the CIB1 transcription 
                     factor from Arabidopsis thaliana"
                     /label=CIBN
                     /note="binds to CRY2 (cryptochrome 2) that has absorbed 
                     blue light (Kennedy et al., 2010)"
                     /translation="MNGAIGGDLLLNFPDMSVLERQRAHLKYLNPTFDSPLAGFFADSS
                     MITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKAAKFDTETKDCN
                     EAAKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKE
                     LEKTDYI"
     CDS             3166..3873
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      3421..3430
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3892..3911)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3957..4012
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4011..4030)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4373..4389)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4373..4389)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4386..4408)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4397..4413
                     /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(4421..4427,4428..4445,4446..4451))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4466..4487
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4546..4741
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4592..4727
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4747..4881
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4784..4803)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4838..4857
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4949..4966)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5120..5708)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5200..5219)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5879..6670,6671..6739))
                     /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     6502..6521
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6740..6844)
                     /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 gggggggggg gcgcgccrgg sggggggggg gggggggggg
      541 gggggggggg ggsgrggsgg rgrggkgcgg 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 gcaggkgggg gtgccgggcg
     1261 gggcggggcc gccycgggcc ggggrgggct cgggggrggg 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 caaagaattc
     1741 accatgaatg gagctatagg aggtgacctt ttgctcaatt ttcctgacat gtcggtccta
     1801 gagcgccaaa gggctcacct caagtacctc aatcccacct ttgattctcc tctcgccggc
     1861 ttctttgccg attcttcaat gattaccggc ggcgagatgg acagctatct ttcgactgcc
     1921 ggtttgaatc ttccgatgat gtacggtgag acgacggtgg aaggtgattc aagactctca
     1981 atttcgccgg aaacgacgct tgggactgga aatttcaagg cagcgaagtt tgatacagag
     2041 actaaggatt gtaatgaggc ggcgaagaag atgacgatga acagagatga cctagtagaa
     2101 gaaggagaag aagagaagtc gaaaataaca gagcaaaaca atgggagcac aaaaagcatc
     2161 aagaagatga aacacaaagc caagaaagaa gagaacaatt tctctaatga ttcatctaaa
     2221 gtgacgaagg aattggagaa aacggattat attcatctcg aggccatgcc ttcccagaaa
     2281 acctaccagg gcagctacgg tttccgtctg ggcttcttgc attctgggac agccaagtct
     2341 gtgacttgca cgtactcccc tgccctcaac aagatgtttt gccaactggc caagacctgc
     2401 cctgtgcagc tgtgggttga ttccacaccc ccgcccggca cccgcgtccg cgccatggcc
     2461 atctacaagc agtcacagca catgacggag gttgtgaggc gctgccccca ccatgagcgc
     2521 tgctcagata gcgatggtct ggcccctcct cagcatctta tccgagtgga aggaaatttg
     2581 cgtgtggagt atttggatga cagaaacact tttcgacata gtgtggtggt gccctatgag
     2641 ccgcctgagg ttggctctga ctgtaccacc atccactaca actacatgtg taacagttcc
     2701 tgcatgggcg gcatgaaccg gaggcccatc ctcaccatca tcacactgga agactccagt
     2761 ggtaatctac tgggacggaa cagctttgag gtgcgtgttt gtgcctgtcc tgggagagac
     2821 cggcgcacag aggaagagaa tctccgcaag aaaggggagc ctcaccacga gctgccccca
     2881 gggagcacta agcgagcact gcccaacaac accagctcct ctccccagcc aaagaagaaa
     2941 ccactggatg gagaatattt cacccttcag atccgtgggc gtgagcgctt cgagatgttc
     3001 cgagagctga atgaggcctt ggaactcaag gatgcccagg ctgggaagga gccagggggg
     3061 agcagggctc actccagcca cctgaagtcc aaaaagggtc agtctacctc ccgccataaa
     3121 aaactcatgt tcaagacaga agggcctgac tcagacggcg gccgcatggt gagcaagggc
     3181 gaggaggata acatggcctc tctcccagcg acacatgagt tacacatctt tggctccatc
     3241 aacggtgtgg actttgacat ggtgggtcag ggcaccggca atccaaatga tggttatgag
     3301 gagttaaacc tgaagtccac caagggtgac ctccagttct ccccctggat tctggtccct
     3361 catatcgggt atggcttcca tcagtacctg ccctaccctg acgggatgtc gcctttccag
     3421 gccgccatgg tagatggctc cggctaccaa gtccatcgca caatgcagtt tgaagatggt
     3481 gcctccctta ctgttaacta ccgctacacc tacgagggaa gccacatcaa aggagaggcc
     3541 caggtgaagg ggactggttt ccctgctgac ggtcctgtga tgaccaactc gctgaccgct
     3601 gcggactggt gcaggtcgaa gaagacttac cccaacgaca aaaccatcat cagtaccttt
     3661 aagtggagtt acaccactgg aaatggcaag cgctaccgga gcactgcgcg gaccacctac
     3721 acctttgcca agccaatggc ggctaactat ctgaagaacc agccgatgta cgtgttccgt
     3781 aagacggagc tcaagcactc caagaccgag ctcaacttca aggagtggca aaaggccttt
     3841 accgatgtga tgggcatgga cgagctgtac aagtctagat aagtcgacgg atctttttcc
     3901 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata
     3961 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     4021 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc
     4081 aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta
     4141 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta
     4201 gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt
     4261 acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc
     4321 tcttctctta tggagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc
     4381 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4441 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4501 cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc
     4561 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4621 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4681 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4741 aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4801 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4861 gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4921 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4981 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5041 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5101 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5161 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5221 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5281 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5341 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5401 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5461 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5521 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5581 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5641 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5701 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5761 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5821 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5881 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5941 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6001 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6061 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6121 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6181 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6241 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6301 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6361 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6421 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6481 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6541 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6601 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6661 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6721 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6781 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6841 cgcgcacatt tccccgaaaa gtgccacctg
//
