LOCUS       sequence-90203-e        6233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian codon optimized CRY2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6233)
  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 6233)
  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-CRY2-mCherry
FEATURES             Location/Qualifiers
     source          1..6233
                     /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      2118..2127
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2124..2831
                     /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(2269..2287)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2537..2556
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2832..2897
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3021..3142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3058..3077)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3112..3131
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3149..3604)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3286..3307)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3498..3517
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3631..3735
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3737..4094
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3737..3928
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3760..3780)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3945..4080
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4129..4923
                     /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             4129..4923
                     /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(4183..4202)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4793..4812
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5111..5130)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5155..5202
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5531..6119
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6020..6039
                     /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 ccatgaagat ggacaaaaag accatcgtct ggtttcggag agatttgaga atagaagata
      661 atcccgcgct cgccgccgcg gcccacgagg gttccgtctt ccccgttttc atttggtgtc
      721 ctgaagaaga aggccagttt tatcccggaa gggcctctag gtggtggatg aagcaaagtc
      781 tggcccatct tagccagtca ctgaaagcac tgggcagtga tcttaccctg atcaagacac
      841 acaataccat ctctgccatc ctcgactgca tcagggtgac cggcgcaacg aaagtcgtgt
      901 ttaaccacct gtacgatcca gttagtctgg tgcgcgacca cactgtgaag gagaagctgg
      961 tggaacgggg gatcagtgtg cagagctaca acggggacct tctgtacgag ccatgggaga
     1021 tctattgcga gaaagggaaa ccgttcacct ccttcaacag ttactggaag aaatgtttgg
     1081 atatgtcaat agagtccgtt atgttgcccc ctccctggag actgatgccg attactgctg
     1141 ctgcagaggc catctgggcc tgctccatcg aggaactcgg tctggaaaat gaagcagaaa
     1201 agccaagcaa tgcacttctc actagagcct ggagccccgg ctggtctaat gccgacaagc
     1261 tgcttaacga gttcatcgaa aaacaactga ttgactacgc gaagaactcc aagaaagtgg
     1321 taggtaactc aactagcttg ctctctccat atctccattt tggcgagatt tctgtccgcc
     1381 atgtatttca gtgcgctcgg atgaaacaga ttatctgggc tcgcgataaa aacagcgaag
     1441 gcgaagaaag cgccgatctg ttcctgcgag ggatcggact tcgggaatac tcccggtata
     1501 tatgtttcaa ctttccattc acacacgagc agagtctgtt gtcccacctc aggttcttcc
     1561 cctgggacgc cgatgtcgac aaattcaagg catggagaca gggaaggaca gggtacccac
     1621 tcgtggatgc tggcatgaga gagctctggg ctacaggctg gatgcacaac cgcatccggg
     1681 taatcgtgtc ctcatttgct gtcaagtttc tgctcctgcc ttggaaatgg ggaatgaagt
     1741 acttttggga tacccttctc gacgccgact tggagtgtga cattctggga tggcaatata
     1801 ttagcgggtc aattcctgac ggccatgagt tggacaggtt ggacaatccg gccttgcagg
     1861 gagctaagta tgatcccgaa ggagagtata ttcgacagtg gctccccgag ctggcccgac
     1921 ttcctacgga gtggattcac catccttggg acgcaccact gacagtgctc aaggcaagcg
     1981 gggtggagct gggcaccaat tacgctaagc ctatagttga tatagataca gcacgcgagc
     2041 tgctggctaa agcgatctct cgcactcggg aggcgcagat tatgatcggt gctgccggca
     2101 gcggcagccc accggtcgcc accatggtga gcaagggcga ggaggataac atggccatca
     2161 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
     2221 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     2281 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
     2341 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     2401 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     2461 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     2521 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     2581 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     2641 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     2701 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     2761 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     2821 agctgtacaa gtccggactc agatctcgag ctcaagcttc gaattctgca gtcgacggta
     2881 ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat accacatttg
     2941 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     3001 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     3061 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3121 ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc
     3181 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     3241 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     3301 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     3361 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     3421 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     3481 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     3541 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     3601 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3661 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3721 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     3781 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3841 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3901 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3961 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4021 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4081 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4141 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4201 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4261 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     4321 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4381 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4441 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4501 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4561 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4621 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     4681 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4741 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4801 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4861 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4921 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4981 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5041 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     5101 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     5161 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     5221 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     5281 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     5341 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     5401 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5461 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5521 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5581 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5641 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     5701 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5761 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5821 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5881 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5941 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6001 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6061 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6121 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6181 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
