LOCUS       sequence-90202-e        5687 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 5687)
  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 5687)
  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-mRFP1-MP
FEATURES             Location/Qualifiers
     source          1..5687
                     /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             1131..1805
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1131..1805
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1261..1279)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1529..1548
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1723..1746
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2475..2596
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2512..2531)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2566..2585
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2603..3058)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2740..2761)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2952..2971
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3085..3189
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3191..3548
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3191..3382
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3214..3234)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3399..3534
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3583..4377
                     /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             3583..4377
                     /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(3637..3656)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4247..4266
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4565..4584)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4609..4656
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4985..5573
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5474..5493
                     /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 atggcctcct
     1141 ccgaggacgt catcaaggag ttcatgcgct tcaaggtgcg catggagggc tccgtgaacg
     1201 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     1261 ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccctc
     1321 agttccagta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     1381 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     1441 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     1501 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     1561 aggcctccac cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagatga
     1621 ggctgaagct gaaggacggc ggccactacg acgccgaggt caagaccacc tacatggcca
     1681 agaagcccgt gcagctgccc ggcgcctaca agaccgacat caagctggac atcacctccc
     1741 acaacgagga ctacaccatc gtggaacagt acgagcgcgc cgagggccgc cactccaccg
     1801 gcgccttgta caagtccgga ctcagatctc gagctcaagc ttcgaattct ggagggaaga
     1861 gtggaggaaa caagaagaac gatggtgtga aggaatcttc tgagagcacc aacaccacca
     1921 ttgaggacga agacaccaaa gtgcgcaaac aggaaattat caaagtgaca gagcagctga
     1981 tcgaagccat aagcaatgga gactttgagt cctacacgaa gatgtgcgac cctggaatga
     2041 cagcctttga accagaggcc ctggggaacc tggtggaggg cctggacttt catcgattct
     2101 attttgaaaa cctgtggtcc cggaacagca agcccgtgca caccaccatc ctgaaccctc
     2161 acatccacct gatgggtgac gagtcagcct gcatcgccta tatccgcatc actcagtacc
     2221 tggatgcagg cggcataccc cgcacggccc agtcagagga gacccgcgtc tggcaccgca
     2281 gggacggcaa atggcagatc gtccacttcc acagatctgg ggcgccctcc gtcctgcccc
     2341 attgaggatc caccggatct agataactga tcataatcag ccataccaca tttgtagagg
     2401 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     2461 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2521 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2581 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     2641 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     2701 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     2761 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     2821 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     2881 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     2941 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     3001 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     3061 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3121 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3181 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     3241 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3301 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3361 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3421 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3481 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3541 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     3601 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3661 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3721 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     3781 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     3841 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     3901 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     3961 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4021 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4081 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     4141 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4201 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4261 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4321 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4381 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4441 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4501 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     4561 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     4621 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     4681 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     4741 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     4801 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     4861 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4921 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4981 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5041 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5101 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     5161 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5221 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5281 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5341 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5401 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5461 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5521 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5581 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5641 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
