LOCUS       sequence-90201-e        5720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For use in light-induced protein inactivation through the 
            interaction with CRY2 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5720)
  AUTHORS   Lee S, Park H, Kyung T, Kim NY, Kim S, Kim J, Heo WD
  TITLE     Reversible protein inactivation by optogenetic trapping in cells.
  JOURNAL   Nat Methods. 2014 Jun;11(6):633-6. doi: 10.1038/nmeth.2940. Epub 
            2014 May 4.
  PUBMED    24793453
REFERENCE   2  (bases 1 to 5720)
  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"; doi: 
            "10.1038/nmeth.2940"; journalName: "Nat Methods"; date: "2014-06"; 
            volume: "11"; issue: "6"; pages: "633-6"
COMMENT     Sequence Label: pCMV-CIB1-mCerulean-MP
FEATURES             Location/Qualifiers
     source          1..5720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             603..1112
                     /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"
     regulatory      1125..1134
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1131..1133,1134..1136,1137..1847)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1176..1197)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1437..1456)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1784..1805
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2508..2629
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2545..2564)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2599..2618
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2636..3091)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2773..2794)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2985..3004
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3118..3222
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3224..3581
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3224..3415
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3247..3267)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3432..3567
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3616..4410
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3616..4410
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3670..3689)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4280..4299
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4598..4617)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4642..4689
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5018..5606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5507..5526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcca
      601 ccatgaatgg agctatagga ggtgaccttt tgctcaattt tcctgacatg tcggtcctag
      661 agcgccaaag ggctcacctc aagtacctca atcccacctt tgattctcct ctcgccggct
      721 tctttgccga ttcttcaatg attaccggcg gcgagatgga cagctatctt tcgactgccg
      781 gtttgaatct tccgatgatg tacggtgaga cgacggtgga aggtgattca agactctcaa
      841 tttcgccgga aacgacgctt gggactggaa atttcaaggc agcgaagttt gatacagaga
      901 ctaaggattg taatgaggcg gcgaagaaga tgacgatgaa cagagatgac ctagtagaag
      961 aaggagaaga agagaagtcg aaaataacag agcaaaacaa tgggagcaca aaaagcatca
     1021 agaagatgaa acacaaagcc aagaaagaag agaacaattt ctctaatgat tcatctaaag
     1081 tgacgaagga attggagaaa acggattata ttcatgtacc ggtcgccacc atggtgagca
     1141 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1201 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     1261 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1321 ccctgacctg gggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     1381 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1441 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1501 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1561 acaacgccat cagcgacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     1621 ccaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     1681 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1741 cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1801 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc ggactcagat
     1861 ctcgagctca agcttcgaat tctggaggga agagtggagg aaacaagaag aacgatggtg
     1921 tgaaggaatc ttctgagagc accaacacca ccattgagga cgaagacacc aaagtgcgca
     1981 aacaggaaat tatcaaagtg acagagcagc tgatcgaagc cataagcaat ggagactttg
     2041 agtcctacac gaagatgtgc gaccctggaa tgacagcctt tgaaccagag gccctgggga
     2101 acctggtgga gggcctggac tttcatcgat tctattttga aaacctgtgg tcccggaaca
     2161 gcaagcccgt gcacaccacc atcctgaacc ctcacatcca cctgatgggt gacgagtcag
     2221 cctgcatcgc ctatatccgc atcactcagt acctggatgc aggcggcata ccccgcacgg
     2281 cccagtcaga ggagacccgc gtctggcacc gcagggacgg caaatggcag atcgtccact
     2341 tccacagatc tggggcgccc tccgtcctgc cccattgagg atccaccgga tctagataac
     2401 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2461 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2521 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2581 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt
     2641 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2701 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2761 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2821 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     2881 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2941 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3001 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3061 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     3121 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3181 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     3241 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3301 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3361 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3421 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3481 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3541 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     3601 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3661 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3721 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3781 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3841 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3901 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3961 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4021 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4081 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4141 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4201 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4261 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4321 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4381 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4441 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4501 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4561 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     4621 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     4681 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     4741 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     4801 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     4861 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4921 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4981 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5041 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5101 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5161 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5221 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5281 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5341 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5401 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5461 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5521 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5581 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5641 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5701 accgtattac cgccatgcat
//
