LOCUS       sequence-299819-        7437 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminal fragment of Cre fused to CRY2 for blue 
            light-induced activation of DNA recombination when used with 
            CIBN-CreC. Also expresses mCherry..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7437)
  AUTHORS   Kennedy MJ, Hughes RM, Peteya LA, Schwartz JW, Ehlers MD, Tucker CL
  TITLE     Rapid blue-light-mediated induction of protein interactions in 
            living cells.
  JOURNAL   Nat Methods. 2010 Dec;7(12):973-5. doi: 10.1038/nmeth.1524
  PUBMED    21037589
REFERENCE   2  (bases 1 to 7437)
  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"; 
            journalName: "Nat Methods"; date: "2010-12"; volume: "7"; issue: 
            "12"; pages: "973-5"
COMMENT     Sequence Label: pmCherry-CRY2-CreN
FEATURES             Location/Qualifiers
     source          1..7437
                     /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"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1320
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(758..776)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1026..1045
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1922..1938)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1923..1939)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             1950..3785
                     /codon_start=1
                     /product="Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAAPDEIVADSFEALGANTIKE
                     PGLCPSVSSNDQQVPSAVRYNGSKRVKPEEEEERDMKKSRGFDERELFSTAESSSSSSV
                     FFVSQSCSLASEGKNLEGIQDSSDQITTSLGKNGCK"
     polyA_signal    4225..4346
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4262..4281)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4316..4335
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4353..4808)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4490..4511)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4702..4721
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4835..4939
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4941..5298
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4941..5132
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4964..4984)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5149..5284
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5333..6127
                     /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             5333..6127
                     /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(5387..5406)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5997..6016
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6315..6334)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6359..6406
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6735..7323
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7224..7243
                     /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 gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
      661 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      721 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
      781 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
      841 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
      901 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      961 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     1021 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     1081 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     1141 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1201 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     1261 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1321 tgtggcggct agtactccgg tattgcggta cccttgtacg cctgttttat actcccttcc
     1381 cgtaacttag acgcacaaaa ccaagttcaa tagaaggggg tacaaaccag taccaccacg
     1441 aacaagcact tctgtttccc cggtgatgtc gtatagactg cttgcgtggt tgaaagcgac
     1501 ggatccgtta tccgcttatg tacttcgaga agcccagtac cacctcggaa tcttcgatgc
     1561 gttgcgctca gcactcaacc ccagagtgta gcttaggctg atgagtctgg acatccctca
     1621 ccggtgacgg tggtccaggc tgcgttggcg gcctacctat ggctaacgcc atgggacgct
     1681 agttgtgaac aaggtgtgaa gagcctattg agctacataa gaatcctccg gcccctgaat
     1741 gcggctaatc ccaacctcgg agcaggtggt cacaaaccag tgattggcct gtcgtaacgc
     1801 gcaagtccgt ggcggaaccg actactttgg gtgtccgtgt ttccttttat tttattgtgg
     1861 ctgcttatgg tgacaatcac agattgttat cataaagcga attggatagg atcaagctta
     1921 tcgataccgt cgacctcgag ctcgccacca tgaagatgga caaaaagact atagtttggt
     1981 ttagaagaga cctaaggatt gaggataatc ctgcattagc agcagctgct cacgaaggat
     2041 ctgtttttcc tgtcttcatt tggtgtcctg aagaagaagg acagttttat cctggaagag
     2101 cttcaagatg gtggatgaaa caatcacttg ctcacttatc tcaatccttg aaggctcttg
     2161 gatctgacct cactttaatc aaaacccaca acacgatttc agcgatcttg gattgtatcc
     2221 gcgttaccgg tgctacaaaa gtcgtcttta accacctcta tgatcctgtt tcgttagttc
     2281 gggaccatac cgtaaaggag aagctggtgg aacgtgggat ctctgtgcaa agctacaatg
     2341 gagatctatt gtatgaaccg tgggagatat actgcgaaaa gggcaaacct tttacgagtt
     2401 tcaattctta ctggaagaaa tgcttagata tgtcgattga atccgttatg cttcctcctc
     2461 cttggcggtt gatgccaata actgcagcgg ctgaagcgat ttgggcgtgt tcgattgaag
     2521 aactagggct ggagaatgag gccgagaaac cgagcaatgc gttgttaact agagcttggt
     2581 ctccaggatg gagcaatgct gataagttac taaatgagtt catcgagaag cagttgatag
     2641 attatgcaaa gaacagcaag aaagttgttg ggaattctac ttcactactt tctccgtatc
     2701 tccatttcgg ggaaataagc gtcagacacg ttttccagtg tgcccggatg aaacaaatta
     2761 tatgggcaag agataagaac agtgaaggag aagaaagtgc agatcttttt cttaggggaa
     2821 tcggtttaag agagtattct cggtatatat gtttcaactt cccgtttact cacgagcaat
     2881 cgttgttgag tcatcttcgg tttttccctt gggatgctga tgttgataag ttcaaggcct
     2941 ggagacaagg caggaccggt tatccgttgg tggatgccgg aatgagagag ctttgggcta
     3001 ccggatggat gcataacaga ataagagtga ttgtttcaag ctttgctgtg aagtttcttc
     3061 tccttccatg gaaatgggga atgaagtatt tctgggatac acttttggat gctgatttgg
     3121 aatgtgacat ccttggctgg cagtatatct ctgggagtat ccccgatggc cacgagcttg
     3181 atcgcttgga caatcccgcg ttacaaggcg ccaaatatga cccagaaggt gagtacataa
     3241 ggcaatggct tcccgagctt gcgagattgc caactgaatg gatccatcat ccatgggacg
     3301 ctcctttaac cgtactcaaa gcttctggtg tggaactcgg aacaaactat gcgaaaccca
     3361 ttgtagacat cgacacagct cgtgagctac tagctaaagc tatttcaaga acccgtgaag
     3421 cacagatcat gatcggagca gcacctgatg agattgtagc agatagcttc gaggccttag
     3481 gggctaatac cattaaagaa cctggtcttt gcccatctgt gtcttctaat gaccaacaag
     3541 taccttcggc tgttcgttac aacgggtcaa agagagtgaa acctgaggaa gaagaagaga
     3601 gagacatgaa gaaatctagg ggattcgatg aaagggagtt gttttcgact gctgaatctt
     3661 cttcttcttc gagtgtgttt ttcgtttcgc agtcttgctc gttggcatca gaagggaaga
     3721 atctggaagg tattcaagat tcatctgatc agattactac aagtttggga aaaaatggtt
     3781 gcaaaggtgg cggtggctct ggaggtggtg ggtccggagg aggcggccgc acgagtgatg
     3841 aggttcgcaa gaacctgatg gacatgttca gggatcgcca ggcgttttct gagcatacct
     3901 ggaaaatgct tctgtccgtt tgccggtcgt gggcggcatg gtgcaagttg aataaccgga
     3961 aatggtttcc cgcagaacct gaagatgttc gcgattatct tctatatctt caggcgcgcg
     4021 gtctggcagt aaaaactatc cagcaacatt tgggccagct aaacatgctt catcgtcggt
     4081 ccgggctgta acccgggatc caccggatct agataactga tcataatcag ccataccaca
     4141 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     4201 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     4261 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4321 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     4381 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     4441 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     4501 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     4561 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     4621 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     4681 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     4741 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     4801 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4861 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4921 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     4981 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5041 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5101 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     5161 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     5221 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5281 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     5341 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     5401 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     5461 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     5521 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     5581 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     5641 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     5701 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     5761 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     5821 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     5881 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     5941 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     6001 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     6061 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     6121 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     6181 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     6241 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     6301 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     6361 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     6421 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     6481 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     6541 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     6601 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6661 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6721 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6781 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6841 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6901 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6961 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7021 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7081 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7141 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7201 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7261 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7321 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7381 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
