LOCUS       sequence-325700-        4029 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector for: CheRiff-eGFP channelrhodopsin (plasma membrane).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4029)
  AUTHORS   Werley CA, Boccardo S, Rigamonti A, Hansson EM, Cohen AE
  TITLE     Multiplexed Optical Sensors in Arrayed Islands of Cells for 
            multimodal recordings of cellular physiology.
  JOURNAL   Nat Commun. 2020 Aug 4;11(1):3881. doi: 10.1038/s41467-020-17607-5.
  PUBMED    32753572
REFERENCE   2  (bases 1 to 4029)
  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"; journalName: "Nat 
            Commun."; date: "2020-08-4"; volume: "11(1):3881. doi"; pages: " 
            10.1038/s41467-020-17607-5"
COMMENT     Sequence Label: pMOS010E: Entry vector for: CheRiff-eGFP
            chann...
FEATURES             Location/Qualifiers
     source          1..4029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(125..713)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(205..224)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(803..1612)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1526..1545
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1604..1623
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    1735..1834
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(join(1864..2577,2578..2580))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(1867..2577,2578..2580))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1909..1930)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2258..2277
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2517..2538
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(2650..3597)
                     /codon_start=1
                     /product="blue light-activated optogenetic actuator derived
                     from Scherffelia dubia channelrhodopsin (Hochbaum et al., 
                     2014)"
                     /label=CheRiff
                     /translation="MGGAPAPDAHSAPPGNDSAGGSEYHAPAGYQVNPPYHPVHGYEEQ
                     CSSIYIYYGALWEQETARGFQWFAVFLSALFLAFYGWHAYKASVGWEEVYVCSVELIKV
                     ILEIYFEFTSPAMLFLYGGNITPWLRYAEWLLTCPVILIHLSNITGLSEAYNKRTMALL
                     VSDLGTICMGVTAALATGWVKWLFYCIGLVYGTQTFYNAGIIYVESYYIMPAGGCKKLV
                     LAMTAVYYSSWLMFPGLFIFGPEGMHTLSVAGSTIGHTIADLLSKNIWGLLGHFLRIKI
                     HEHIIMYGDIRRPVSSQFLGRKVDVLAFVTEEDKV"
     regulatory      3594..3603
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3635..3734)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3753..3776)
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     terminator      complement(3784..3811)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3903..3989)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ccccgagatc catgctagcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
       61 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      121 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      181 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      241 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      301 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
      361 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
      421 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
      481 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
      541 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
      601 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
      661 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
      721 ctttgatctt ttctacgggg tctgacgctc agtggaacgg ggcccaatct gaataatgtt
      781 acaaccaatt aaccaattct gattagaaaa actcatcgag catcaaatga aactgcaatt
      841 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
      901 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
      961 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     1021 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     1081 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     1141 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     1201 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     1261 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttt ccggggatcg
     1321 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     1381 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     1441 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aagcgataga
     1501 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     1561 ccatgttgga atttaatcgc ggcctcgacg tttcccgttg aatatggctc ataacacccc
     1621 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt
     1681 gtgcaatgta acatcagaga ttttgagaca cgggccagag ctgcagctgg atggcaaata
     1741 atgattttat tttgactgat agtgacctgt tcgttgcaac aaattgataa gcaatgcttt
     1801 cttataatgc caactttgta caagaaagct gggtctagat atctcgagtg cggccgcgaa
     1861 ttcttacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag
     1921 caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact gggtgctcag
     1981 gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg
     2041 gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat
     2101 gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag
     2161 cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt
     2221 caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt
     2281 gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg
     2341 ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca gggtggtcac
     2401 gagggtgggc cagggcacgg gcagcttgcc ggtggtgcaa atgaacttca gggtcagctt
     2461 gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc
     2521 gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac
     2581 cacgttgatg tcgatctggt ccagggggat gtactcgccc tcgctggtga tcctgctctt
     2641 ggcggccgcc actttatctt cctctgtcac gaaggccagc acgtccacct tgcgtcccag
     2701 aaactgggag ctcactggtc tcctgatatc gccgtacata atgatatgct cgtgaatttt
     2761 gatccgcagg aagtgcccca gcagtcccca aatattcttg gacagcaggt cggcgatggt
     2821 atggccaata gtggacccag ccacgctcag ggtgtgcatg ccttcaggcc caaagatgaa
     2881 caggccggga aacatcagcc aactagaata gtacacggca gtcatggcca gcaccagttt
     2941 cttacagccg ccggcaggca tgatatagta agactccacg tagatgattc cagcgttgta
     3001 gaatgtctgg gttccataca ccaggccgat acagtaaaac agccacttca cccacccagt
     3061 ggccagagcg gctgtcactc ccatgcaaat agttcccagg tcggacacca gcagagccat
     3121 tgtccgctta ttgtatgcct cactcaggcc ggtgatgtta gacagatgaa tcaggatcac
     3181 gggacatgtc agcagccatt cggcatatct cagccatggg gtaatgttcc ctccgtacag
     3241 gaacagcata gcaggactgg tgaactcgaa atagatctcc agaatcactt tgatcagctc
     3301 cacggagcac acgtacactt cctcccatcc cacgctggcc ttataggcgt gccagccgta
     3361 gaaagccaga aacagggcag acaggaacac ggcaaaccac tggaagcccc tagctgtttc
     3421 ctgctcccac agggccccat agtagatgta gatggagctg cactgttcct catacccatg
     3481 cacggggtgg tagggtggat tcacttgata tccagctggg gcatggtact cactgcctcc
     3541 ggcagaatcg tttccaggtg gggcgctgtg agcgtctgga gcaggagctc cgcccatggt
     3601 ggcggatcca gtcgactgaa ttggttccgg cgccagcctg cttttttgta caaagttggc
     3661 attataaaaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
     3721 aaaatcatta tttggggccc gagcttaagt aactaactaa caggaagagt ttgtagaaac
     3781 gcaaaaaggc catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc
     3841 gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc
     3901 ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag
     3961 tcttccgact gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcttagta
     4021 gttagacgt
//
