LOCUS       sequence-198851-        7357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses fusion of CRY2PHR with mCherry and LAMP1 (1-415).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7357)
  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 7357)
  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: 
            "10.1016/j.chembiol.2015.04.014"; journalName: "Chem Biol"; date: 
            "2015-05-21- 21"; volume: "22"; issue: "5"; pages: "671-82"
COMMENT     Sequence Label: LAMP-mCherry-CRY2
FEATURES             Location/Qualifiers
     source          1..7357
                     /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      618..627
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1884..1893
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1890..2597
                     /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(2035..2053)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2303..2322
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2634..4127
                     /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    4145..4266
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4182..4201)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4236..4255
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4273..4728)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4410..4431)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4622..4641
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4755..4859
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4861..5218
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4861..5052
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4884..4904)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5069..5204
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5253..6047
                     /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             5253..6047
                     /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(5307..5326)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5917..5936
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6235..6254)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6279..6326
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6655..7243
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7144..7163
                     /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 atctcgagcc accatggcgc cccgcagcgc ccggcgaccc ctgctgctgc
      661 tactgctgtt gctgctgctc ggcctcatgc attgtgcgtc agcagcaatg tttatggtga
      721 aaaatggcaa cgggaccgcg tgcataatgg ccaacttctc tgctgccttc tcagtgaact
      781 acgacaccaa gagtggccct aagaacatga cctttgacct gccatcagat gccacagtgg
      841 tgctcaaccg cagctcctgt ggaaaagaga acacttctga ccccagtctc gtgattgctt
      901 ttggaagagg acatacactc actctcaatt tcacgagaaa tgcaacacgt tacagcgtcc
      961 agctcatgag ttttgtttat aacttgtcag acacacacct tttccccaat gcgagctcca
     1021 aagaaatcaa gactgtggaa tctataactg acatcagggc agatatagat aaaaaataca
     1081 gatgtgttag tggcacccag gtccacatga acaacgtgac cgtaacgctc catgatgcca
     1141 ccatccaggc gtacctttcc aacagcagct tcagccgggg agagacacgc tgtgaacaag
     1201 acaggccttc cccaaccaca gcgccccctg cgccacccag cccctcgccc tcacccgtgc
     1261 ccaagagccc ctctgtggac aagtacaacg tgaacggcac caacgggacc tgcctgctgg
     1321 ccagcatggg gctgcagctg aacctcacct atgagaggaa ggacaacacg acggtgacaa
     1381 ggcttctcaa catcaacccc aacaagacct cggccagcgg gagctgcggc gcccacctgg
     1441 tgactctgga gctgcacagc gagggcacca ccgtcctgct cttccagttc gggatgaatg
     1501 caagttctag ccggtttttc ctacaaggaa tccagttgaa tacaattctt cctgacgcca
     1561 gagaccctgc ctttaaagct gccaacggct ccctgcgagc gctgcaggcc acagtcggca
     1621 attcctacaa gtgcaacgcg gaggagcacg tccgtgtcac gaaggcgttt tcagtcaata
     1681 tattcaaagt gtgggtccag gctttcaagg tggaaggtgg ccagtttggc tctgtggagg
     1741 agtgtctgct ggacgagaac agcacgctga tccccatcgc tgtgggtggt gccctggcgg
     1801 ggctggtcct catcgtcctc atcgcctacc tcgtcggcag gaagaggagg cacgcaggct
     1861 accagactat ggatccaccg gtcgccacca tggtgagcaa gggcgaggag gataacatgg
     1921 ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg
     1981 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     2041 tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca
     2101 tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt
     2161 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga
     2221 ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg
     2281 gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct
     2341 cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga
     2401 agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc
     2461 ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg
     2521 aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca
     2581 tggacgagct gtacaagggc agtagcggaa gcagtggttc ttcaggatca tccatgaaga
     2641 tggacaaaaa gactatagtt tggtttagaa gagacctaag gattgaggat aatcctgcat
     2701 tagcagcagc tgctcacgaa ggatctgttt ttcctgtctt catttggtgt cctgaagaag
     2761 aaggacagtt ttatcctgga agagcttcaa gatggtggat gaaacaatca cttgctcact
     2821 tatctcaatc cttgaaggct cttggatctg acctcacttt aatcaaaacc cacaacacga
     2881 tttcagcgat cttggattgt atccgcgtta ccggtgctac aaaagtcgtc tttaaccacc
     2941 tctatgatcc tgtttcgtta gttcgggacc ataccgtaaa ggagaagctg gtggaacgtg
     3001 ggatctctgt gcaaagctac aatggagatc tattgtatga accgtgggag atatactgcg
     3061 aaaagggcaa accttttacg agtttcaatt cttactggaa gaaatgctta gatatgtcga
     3121 ttgaatccgt tatgcttcct cctccttggc ggttgatgcc aataactgca gcggctgaag
     3181 cgatttgggc gtgttcgatt gaagaactag ggctggagaa tgaggccgag aaaccgagca
     3241 atgcgttgtt aactagagct tggtctccag gatggagcaa tgctgataag ttactaaatg
     3301 agttcatcga gaagcagttg atagattatg caaagaacag caagaaagtt gttgggaatt
     3361 ctacttcact actttctccg tatctccatt tcggggaaat aagcgtcaga cacgttttcc
     3421 agtgtgcccg gatgaaacaa attatatggg caagagataa gaacagtgaa ggagaagaaa
     3481 gtgcagatct ttttcttagg ggaatcggtt taagagagta ttctcggtat atatgtttca
     3541 acttcccgtt tactcacgag caatcgttgt tgagtcatct tcggtttttc ccttgggatg
     3601 ctgatgttga taagttcaag gcctggagac aaggcaggac cggttatccg ttggtggatg
     3661 ccggaatgag agagctttgg gctaccggat ggatgcataa cagaataaga gtgattgttt
     3721 caagctttgc tgtgaagttt cttctccttc catggaaatg gggaatgaag tatttctggg
     3781 atacactttt ggatgctgat ttggaatgtg acatccttgg ctggcagtat atctctggga
     3841 gtatccccga tggccacgag cttgatcgct tggacaatcc cgcgttacaa ggcgccaaat
     3901 atgacccaga aggtgagtac ataaggcaat ggcttcccga gcttgcgaga ttgccaactg
     3961 aatggatcca tcatccatgg gacgctcctt taaccgtact caaagcttct ggtgtggaac
     4021 tcggaacaaa ctatgcgaaa cccattgtag acatcgacac agctcgtgag ctactagcta
     4081 aagctatttc aagaacccgt gaagcacaga tcatgatcgg agcagcataa caattgttgt
     4141 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4201 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4261 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     4321 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     4381 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     4441 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4501 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     4561 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     4621 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     4681 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     4741 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4801 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     4861 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4921 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4981 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5041 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     5101 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     5161 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     5221 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5281 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5341 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5401 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     5461 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5521 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5581 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5641 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5701 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5761 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5821 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5881 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5941 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     6001 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     6061 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     6121 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     6181 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     6241 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     6301 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     6361 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     6421 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     6481 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     6541 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6601 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6661 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6721 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6781 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6841 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6901 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6961 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7021 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7081 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7141 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7201 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7261 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7321 ccctgattct gtggataacc gtattaccgc catgcat
//
