LOCUS       Exported                5773 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses enhanced sodium pump for light-driven extrusion of sodium;
            activation with green light; codon-optimized for human/mouse.
ACCESSION   .
VERSION     .
KEYWORDS    pEYFP-N1-eKR2
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 May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5773
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    245..301
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      302..311
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1349..2062
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1391..1412)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1652..1671)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1999..2020
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2197..2318
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2234..2253)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2288..2307
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2325..2780)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2462..2483)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2674..2693
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2807..2911
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2913..3270
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2936..2956)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3121..3256
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3183..3202
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3305..4099
                     /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(3359..3378)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3969..3988
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4287..4306)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4331..4378
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4707..5295
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5196..5215
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5470..5773
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc
      301 cgccaccatg gactatgggg gagcactgtc cgccgtcggg ctgtttcaga cttcctacac
      361 cctggagaac aacgggagcg tcatttgcat tcctaacaat ggccagtgct tctgtctggc
      421 ctggctgaag agcaacggga cccaggaact gggcaatgcc aactttgaga atttcatcgg
      481 cgccaccgag ggctttagcg agatcgccta tcagttcacc tcccacatcc tgacactggg
      541 ctatgccgtg atgctggccg gcctgctgta ctttatcctg acaatcaaga acgtggacaa
      601 gaagttccag atgtctaata tcctgagcgc cgtggtcatg gtgagcgcct tcctgctgct
      661 gtatgcccag gcccagaact ggaccagctc ctttacattc aatgaggaag tgggcaggta
      721 ctttctggac ccatccggcg atctgttcaa caatggctac agatatctga actggctgat
      781 cgatgtgccc atgctgctgt ttcagatcct gttcgtggtg tccctgacca catctaagtt
      841 ctctagcgtg cggaaccagt tttggttctc cggcgccatg atgatcatca ccggctatat
      901 cggccagttt tacgaggtgt ctaatctgac agccttcctg gtgtggggcg ccatctcctc
      961 tgccttcttt ttccacatcc tgtgggtcat gaagaaagtg atcaacgagg gcaaggaggg
     1021 aatctctcca gcaggccaga agatcctgag caacatctgg attctgttcc tgatcagctg
     1081 gaccctgtac cccggcgcct atctgatgcc ttacctgaca ggcgtggacg gcttcctgta
     1141 ctccgaggat ggcgtgatgg caaggcagct ggtgtatacc atcgccgacg tgagctccaa
     1201 agtgatctac ggcgtgctgc tgggcaatct ggcaatcacc ctgtcaaaaa acaaggaact
     1261 ggtcgaggca aatagcgcag caccggtaaa gagcaggatc accagcgagg gcgagtacat
     1321 ccccctggac cagatcgaca tcaacgtggt gagcaagggc gaggagctgt tcaccggggt
     1381 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     1441 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     1501 caagctgccc gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt
     1561 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     1621 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     1681 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     1741 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     1801 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     1861 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     1921 ccccgtgctg ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc
     1981 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     2041 cggcatggac gagctgtaca agttctgcta cgagaacgag gtgtgatcta gatcataatc
     2101 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     2161 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2221 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2281 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     2341 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     2401 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     2461 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     2521 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     2581 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     2641 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     2701 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     2761 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2821 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2881 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     2941 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3001 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3061 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3121 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3181 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3241 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     3301 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3361 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3421 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3481 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3541 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3601 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3661 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3721 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3781 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     3841 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3901 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     3961 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4021 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4081 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4141 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4201 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4261 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4321 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     4381 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     4441 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     4501 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     4561 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4621 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4681 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4741 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4801 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4861 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4921 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4981 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5041 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5101 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5161 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5221 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5281 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5341 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5401 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     5461 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     5521 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     5581 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     5641 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     5701 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     5761 cgctattacc atg
//
