LOCUS       sequence-350815-        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"
     promoter        47..404
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        47..238
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(70..90)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      255..390
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             439..1233
                     /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             439..1233
                     /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(493..512)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1103..1122
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1421..1440)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1465..1512
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1841..2429
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2330..2349
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2604..2907
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2908..3111
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3062..3082
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4661..4670
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4667..5374
                     /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(4812..4830)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5080..5099
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    5375..5440
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    5564..5685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5601..5620)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5655..5674
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5692..6147)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5829..5850)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6041..6060
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(6174..6233,1..45)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
       61 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
      121 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
      181 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
      241 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
      301 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
      361 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
      421 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
      481 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
      541 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
      601 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
      661 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
      721 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
      781 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
      841 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
      901 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
      961 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     1021 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     1081 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     1141 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     1201 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     1261 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     1321 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     1381 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     1441 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     1501 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     1561 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     1621 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     1681 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     1741 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1801 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1861 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1921 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1981 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2041 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2101 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2161 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2221 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2281 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2341 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2401 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2461 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2521 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     2581 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     2641 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     2701 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     2761 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     2821 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     2881 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     2941 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     3001 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     3061 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
     3121 aaccgtcaga tccgctagcg ccaccatgaa gatggacaaa aagaccatcg tctggtttcg
     3181 gagagatttg agaatagaag ataatcccgc gctcgccgcc gcggcccacg agggttccgt
     3241 cttccccgtt ttcatttggt gtcctgaaga agaaggccag ttttatcccg gaagggcctc
     3301 taggtggtgg atgaagcaaa gtctggccca tcttagccag tcactgaaag cactgggcag
     3361 tgatcttacc ctgatcaaga cacacaatac catctctgcc atcctcgact gcatcagggt
     3421 gaccggcgca acgaaagtcg tgtttaacca cctgtacgat ccagttagtc tggtgcgcga
     3481 ccacactgtg aaggagaagc tggtggaacg ggggatcagt gtgcagagct acaacgggga
     3541 ccttctgtac gagccatggg agatctattg cgagaaaggg aaaccgttca cctccttcaa
     3601 cagttactgg aagaaatgtt tggatatgtc aatagagtcc gttatgttgc cccctccctg
     3661 gagactgatg ccgattactg ctgctgcaga ggccatctgg gcctgctcca tcgaggaact
     3721 cggtctggaa aatgaagcag aaaagccaag caatgcactt ctcactagag cctggagccc
     3781 cggctggtct aatgccgaca agctgcttaa cgagttcatc gaaaaacaac tgattgacta
     3841 cgcgaagaac tccaagaaag tggtaggtaa ctcaactagc ttgctctctc catatctcca
     3901 ttttggcgag atttctgtcc gccatgtatt tcagtgcgct cggatgaaac agattatctg
     3961 ggctcgcgat aaaaacagcg aaggcgaaga aagcgccgat ctgttcctgc gagggatcgg
     4021 acttcgggaa tactcccggt atatatgttt caactttcca ttcacacacg agcagagtct
     4081 gttgtcccac ctcaggttct tcccctggga cgccgatgtc gacaaattca aggcatggag
     4141 acagggaagg acagggtacc cactcgtgga tgctggcatg agagagctct gggctacagg
     4201 ctggatgcac aaccgcatcc gggtaatcgt gtcctcattt gctgtcaagt ttctgctcct
     4261 gccttggaaa tggggaatga agtacttttg ggataccctt ctcgacgccg acttggagtg
     4321 tgacattctg ggatggcaat atattagcgg gtcaattcct gacggccatg agttggacag
     4381 gttggacaat ccggccttgc agggagctaa gtatgatccc gaaggagagt atattcgaca
     4441 gtggctcccc gagctggccc gacttcctac ggagtggatt caccatcctt gggacgcacc
     4501 actgacagtg ctcaaggcaa gcggggtgga gctgggcacc aattacgcta agcctatagt
     4561 tgatatagat acagcacgcg agctgctggc taaagcgatc tctcgcactc gggaggcgca
     4621 gattatgatc ggtgctgccg gcagcggcag cccaccggtc gccaccatgg tgagcaaggg
     4681 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     4741 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     4801 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     4861 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     4921 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     4981 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     5041 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     5101 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     5161 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     5221 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     5281 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     5341 ccactccacc ggcggcatgg acgagctgta caagtccgga ctcagatctc gagctcaagc
     5401 ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc accggatcta gataactgat
     5461 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     5521 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     5581 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5641 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa
     5701 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     5761 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     5821 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     5881 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     5941 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     6001 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     6061 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     6121 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     6181 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg aga
//
