LOCUS       sequence-198852-        6389 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses fusion of CRY2PHR with mCherry and PEX3 (1-42).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6389)
  AUTHORS   Duan L, Che D, Zhang K, Ong Q, Guo S, Cui B
  TITLE     Optogenetic control of molecular motors and organelle distributions 
            in cells.
  JOURNAL   Chem Biol. 2015 May 21;22(5):671-82. doi: 
            10.1016/j.chembiol.2015.04.014. Epub 2015 May 9.
  PUBMED    25963241
REFERENCE   2  (bases 1 to 6389)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.chembiol.2015.04.014"; journalName: "Chem Biol"; date: 
            "2015-05-21- 21"; volume: "22"; issue: "5"; pages: "671-82"
COMMENT     Sequence Label: PEX-mCherry-CRY2
FEATURES             Location/Qualifiers
     source          1..6389
                     /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             802..819
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             823..855
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(859..867,868..882)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      916..925
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             922..1629
                     /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(1067..1085)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1335..1354
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1666..3159
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     polyA_signal    3177..3298
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3214..3233)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3268..3287
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3305..3760)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3442..3463)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3654..3673
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3787..3891
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3893..4250
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3893..4084
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3916..3936)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4101..4236
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4285..5079
                     /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             4285..5079
                     /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(4339..4358)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4949..4968
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5267..5286)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5311..5358
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5687..6275
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6176..6195
                     /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 ccggactcag atctcgagct caagcttcga attctgcagt cgacggccac catgctgagg
      661 tctgtatgga attttctgaa acgccacaaa aagaaatgca tcttcctggg cacggtcctt
      721 ggaggagtat atattctggg gaaatatgga cagaagaaaa tcagagaaat acaggaaagg
      781 gaaccggtca tgcggggttc tcatcatcat catcatcatg gtatggctag catgactggt
      841 ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggatccggt accgcgggcc
      901 cgggatccac cggtcgccac catggtgagc aagggcgagg aggataacat ggccatcatc
      961 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
     1021 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
     1081 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
     1141 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
     1201 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     1261 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
     1321 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag
     1381 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
     1441 gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag
     1501 ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac
     1561 accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag
     1621 ctgtacaagg gcagtagcgg aagcagtggt tcttcaggat catccatgaa gatggacaaa
     1681 aagactatag tttggtttag aagagaccta aggattgagg ataatcctgc attagcagca
     1741 gctgctcacg aaggatctgt ttttcctgtc ttcatttggt gtcctgaaga agaaggacag
     1801 ttttatcctg gaagagcttc aagatggtgg atgaaacaat cacttgctca cttatctcaa
     1861 tccttgaagg ctcttggatc tgacctcact ttaatcaaaa cccacaacac gatttcagcg
     1921 atcttggatt gtatccgcgt taccggtgct acaaaagtcg tctttaacca cctctatgat
     1981 cctgtttcgt tagttcggga ccataccgta aaggagaagc tggtggaacg tgggatctct
     2041 gtgcaaagct acaatggaga tctattgtat gaaccgtggg agatatactg cgaaaagggc
     2101 aaacctttta cgagtttcaa ttcttactgg aagaaatgct tagatatgtc gattgaatcc
     2161 gttatgcttc ctcctccttg gcggttgatg ccaataactg cagcggctga agcgatttgg
     2221 gcgtgttcga ttgaagaact agggctggag aatgaggccg agaaaccgag caatgcgttg
     2281 ttaactagag cttggtctcc aggatggagc aatgctgata agttactaaa tgagttcatc
     2341 gagaagcagt tgatagatta tgcaaagaac agcaagaaag ttgttgggaa ttctacttca
     2401 ctactttctc cgtatctcca tttcggggaa ataagcgtca gacacgtttt ccagtgtgcc
     2461 cggatgaaac aaattatatg ggcaagagat aagaacagtg aaggagaaga aagtgcagat
     2521 ctttttctta ggggaatcgg tttaagagag tattctcggt atatatgttt caacttcccg
     2581 tttactcacg agcaatcgtt gttgagtcat cttcggtttt tcccttggga tgctgatgtt
     2641 gataagttca aggcctggag acaaggcagg accggttatc cgttggtgga tgccggaatg
     2701 agagagcttt gggctaccgg atggatgcat aacagaataa gagtgattgt ttcaagcttt
     2761 gctgtgaagt ttcttctcct tccatggaaa tggggaatga agtatttctg ggatacactt
     2821 ttggatgctg atttggaatg tgacatcctt ggctggcagt atatctctgg gagtatcccc
     2881 gatggccacg agcttgatcg cttggacaat cccgcgttac aaggcgccaa atatgaccca
     2941 gaaggtgagt acataaggca atggcttccc gagcttgcga gattgccaac tgaatggatc
     3001 catcatccat gggacgctcc tttaaccgta ctcaaagctt ctggtgtgga actcggaaca
     3061 aactatgcga aacccattgt agacatcgac acagctcgtg agctactagc taaagctatt
     3121 tcaagaaccc gtgaagcaca gatcatgatc ggagcagcat aacaattgtt gttgttaact
     3181 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3241 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     3301 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     3361 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     3421 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     3481 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     3541 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     3601 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     3661 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     3721 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     3781 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3841 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     3901 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3961 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4021 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4081 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4141 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     4201 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     4261 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4321 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4381 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4441 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     4501 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4561 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4621 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4681 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4741 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4801 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4861 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4921 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4981 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5041 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5101 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5161 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5221 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     5281 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     5341 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     5401 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     5461 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     5521 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5581 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5641 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5701 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5761 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5821 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5881 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5941 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6001 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6061 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6121 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6181 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6241 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6301 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6361 ctgtggataa ccgtattacc gccatgcat
//
