LOCUS       sequence-217461-        5773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses enhanced sodium pump for light-driven extrusion of sodium;
            activation with green light; codon-optimized for human/mouse.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5773)
  AUTHORS   Grimm C, Silapetere A, Vogt A, Bernal Sierra YA, Hegemann P
  TITLE     Electrical properties, substrate specificity and optogenetic 
            potential of the engineered light-driven sodium pump eKR2.
  JOURNAL   Sci Rep. 2018 Jun 18;8(1):9316. doi: 10.1038/s41598-018-27690-w.
  PUBMED    29915394
REFERENCE   2  (bases 1 to 5773)
  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: "Sci Rep."; 
            date: "2018-06-18"; volume: "8(1):9316. doi"; pages: " 
            10.1038/s41598-018-27690-w"
COMMENT     Sequence Label: pEYFP-N1-eKR2
FEATURES             Location/Qualifiers
     source          1..5773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        53..356
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        357..560
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     511..531
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    601..657
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      658..667
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1705..1707,1708..2418)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1747..1768)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2008..2027)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2355..2376
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2553..2674
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2590..2609)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2644..2663
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2681..3136)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2818..2839)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3030..3049
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3163..3267
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3269..3626
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3269..3460
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3292..3312)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3477..3612
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3661..4455
                     /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             3661..4455
                     /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(3715..3734)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4325..4344
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4643..4662)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4687..4734
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5063..5651
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5552..5571
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
       61 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      121 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      181 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      241 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      301 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      361 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      421 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      481 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      541 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggactc
      601 agatctcgag ctcaagcttc gaattctgca gtcgacggta ccgcgggccc gggatccgcc
      661 accatggact atgggggagc actgtccgcc gtcgggctgt ttcagacttc ctacaccctg
      721 gagaacaacg ggagcgtcat ttgcattcct aacaatggcc agtgcttctg tctggcctgg
      781 ctgaagagca acgggaccca ggaactgggc aatgccaact ttgagaattt catcggcgcc
      841 accgagggct ttagcgagat cgcctatcag ttcacctccc acatcctgac actgggctat
      901 gccgtgatgc tggccggcct gctgtacttt atcctgacaa tcaagaacgt ggacaagaag
      961 ttccagatgt ctaatatcct gagcgccgtg gtcatggtga gcgccttcct gctgctgtat
     1021 gcccaggccc agaactggac cagctccttt acattcaatg aggaagtggg caggtacttt
     1081 ctggacccat ccggcgatct gttcaacaat ggctacagat atctgaactg gctgatcgat
     1141 gtgcccatgc tgctgtttca gatcctgttc gtggtgtccc tgaccacatc taagttctct
     1201 agcgtgcgga accagttttg gttctccggc gccatgatga tcatcaccgg ctatatcggc
     1261 cagttttacg aggtgtctaa tctgacagcc ttcctggtgt ggggcgccat ctcctctgcc
     1321 ttctttttcc acatcctgtg ggtcatgaag aaagtgatca acgagggcaa ggagggaatc
     1381 tctccagcag gccagaagat cctgagcaac atctggattc tgttcctgat cagctggacc
     1441 ctgtaccccg gcgcctatct gatgccttac ctgacaggcg tggacggctt cctgtactcc
     1501 gaggatggcg tgatggcaag gcagctggtg tataccatcg ccgacgtgag ctccaaagtg
     1561 atctacggcg tgctgctggg caatctggca atcaccctgt caaaaaacaa ggaactggtc
     1621 gaggcaaata gcgcagcacc ggtaaagagc aggatcacca gcgagggcga gtacatcccc
     1681 ctggaccaga tcgacatcaa cgtggtgagc aagggcgagg agctgttcac cggggtggtg
     1741 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1801 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1861 ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgca gtgcttcgcc
     1921 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1981 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2041 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2101 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2161 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     2221 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2281 gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac
     2341 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2401 atggacgagc tgtacaagtt ctgctacgag aacgaggtgt gatctagatc ataatcagcc
     2461 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     2521 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     2581 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2641 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     2701 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     2761 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     2821 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     2881 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     2941 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3001 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3061 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3121 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3181 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3241 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     3301 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3361 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3421 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3481 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3541 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3601 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     3661 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3721 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3781 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3841 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3901 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3961 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4021 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4081 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4141 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     4201 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4261 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4321 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4381 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4441 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4501 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4561 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4621 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     4681 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     4741 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     4801 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     4861 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     4921 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4981 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5041 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5101 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5161 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     5221 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5281 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5341 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5401 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5461 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5521 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5581 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5641 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5701 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     5761 tgcattagtt att
//
