LOCUS       sequence-326606-        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"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      489..498
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             495..1442
                     /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"
     CDS             join(1512..1514,1515..2228)
                     /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             join(1512..1514,1515..2225)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1554..1575)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1815..1834)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2162..2183
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2258..2357)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2469..2488)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2480..3289
                     /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     complement(2547..2566)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3379..3967
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3868..3887
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggctggc gccggaacca attcagtcga
      481 ctggatccgc caccatgggc ggagctcctg ctccagacgc tcacagcgcc ccacctggaa
      541 acgattctgc cggaggcagt gagtaccatg ccccagctgg atatcaagtg aatccaccct
      601 accaccccgt gcatgggtat gaggaacagt gcagctccat ctacatctac tatggggccc
      661 tgtgggagca ggaaacagct aggggcttcc agtggtttgc cgtgttcctg tctgccctgt
      721 ttctggcttt ctacggctgg cacgcctata aggccagcgt gggatgggag gaagtgtacg
      781 tgtgctccgt ggagctgatc aaagtgattc tggagatcta tttcgagttc accagtcctg
      841 ctatgctgtt cctgtacgga gggaacatta ccccatggct gagatatgcc gaatggctgc
      901 tgacatgtcc cgtgatcctg attcatctgt ctaacatcac cggcctgagt gaggcataca
      961 ataagcggac aatggctctg ctggtgtccg acctgggaac tatttgcatg ggagtgacag
     1021 ccgctctggc cactgggtgg gtgaagtggc tgttttactg tatcggcctg gtgtatggaa
     1081 cccagacatt ctacaacgct ggaatcatct acgtggagtc ttactatatc atgcctgccg
     1141 gcggctgtaa gaaactggtg ctggccatga ctgccgtgta ctattctagt tggctgatgt
     1201 ttcccggcct gttcatcttt gggcctgaag gcatgcacac cctgagcgtg gctgggtcca
     1261 ctattggcca taccatcgcc gacctgctgt ccaagaatat ttggggactg ctggggcact
     1321 tcctgcggat caaaattcac gagcatatca ttatgtacgg cgatatcagg agaccagtga
     1381 gctcccagtt tctgggacgc aaggtggacg tgctggcctt cgtgacagag gaagataaag
     1441 tggcggccgc caagagcagg atcaccagcg agggcgagta catccccctg gaccagatcg
     1501 acatcaacgt ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1561 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1621 ccacctacgg caagctgacc ctgaagttca tttgcaccac cggcaagctg cccgtgccct
     1681 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     1741 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1801 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1861 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1921 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     1981 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     2041 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     2101 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     2161 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     2221 acaagtaaga attcgcggcc gcactcgaga tatctagacc cagctttctt gtacaaagtt
     2281 ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa
     2341 aataaaatca ttatttgcca tccagctgca gctctggccc gtgtctcaaa atctctgatg
     2401 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     2461 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     2521 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     2581 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     2641 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     2701 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     2761 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     2821 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     2881 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     2941 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     3001 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     3061 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     3121 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     3181 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     3241 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     3301 taattggttg taacattatt cagattgggc cccgttccac tgagcgtcag accccgtaga
     3361 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3421 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3481 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3541 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3601 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3661 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3721 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3781 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3841 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3901 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3961 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4021 tcacatgtt
//
