LOCUS       sequence-469989-        7474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express ChR2 in mitochondria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7474)
  AUTHORS   Ernst P, Xu N, Qu J, Chen H, Goldberg MS, Darley-Usmar V, Zhang JJ, 
            O'Rourke B, Liu X, Zhou L
  TITLE     Precisely Control Mitochondria with Light to Manipulate Cell Fate 
            Decision.
  JOURNAL   Biophys J. 2019 Aug 20;117(4):631-645. doi: 
            10.1016/j.bpj.2019.06.038. Epub 2019 Jul 26.
  PUBMED    31400914
REFERENCE   2  (bases 1 to 7474)
  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: 
            "10.1016/j.bpj.2019.06.038"; journalName: "Biophys J"; date: 
            "2019-08-20- 20"; volume: "117"; issue: "4"; pages: "631-645"
COMMENT     Sequence Label: pCMV-ABCB10-ChR2-eYFP
FEATURES             Location/Qualifiers
     source          1..7474
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1327..2253
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      2263..2272
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2269..2271,2272..2274,2275..2985)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2314..2335)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2575..2594)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2922..2943
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(3027..3045)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3028..3045)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3065..3082)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3071..3295
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3341..3769
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3428..3447)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3638..3659
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3778..3798)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3783..4112
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3963..4098
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4025..4044
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4179..4973
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4233..4252)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4843..4862
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5147..5268
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5184..5203)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5238..5257
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5317..5333)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5317..5333)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5330..5352)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5341..5357
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5365..5371,5372..5389,5390..5395))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5410..5431
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5548..5565)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5719..6307)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5799..5818)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6478..7269,7270..7338))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7101..7120
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7339..7443)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
      901 atgcgcgccc cttctgctag ggcgctactg ctgattccgc gtcggggccc tgccgtgcga
      961 gcgtgggccc cggccgtctc ctctcggata tggctggctt ctgaatggac cccgctcgta
     1021 cgcgcgtgga cctctctgat ccacaagccg ggttcgggcc tccgctttcc cgcgccccta
     1081 tccgggctgc ctggcggcgt ggggcagtgg gccacctcct cgggggcccg caggtgctgg
     1141 gtgctggcag gaccccgcgc cgcacatccc ctgttcgcca ggctccaggg tgcagctgcc
     1201 accggtgtgc gagaccttgg gaacgactcg cagcggcgtc ccgcggcgac cgggcgctca
     1261 gaagtatgga agctcctagg gctggtgcgc cccgagcgcg ggagactgtc agctgcagtt
     1321 gtcgacatgg actatggcgg cgctttgtct gccgtcggac gcgaactttt gttcgttact
     1381 aatcctgtgg tggtgaacgg gtccgtcctg gtccctgagg atcaatgtta ctgtgccgga
     1441 tggattgaat ctcgcggcac gaacggcgct cagaccgcgt caaatgtcct gcagtggctt
     1501 gcagcaggat tcagcatttt gctgctgatg ttctatgcct accaaacctg gaaatctaca
     1561 tgcggctggg aggagatcta tgtgtgcgcc attgaaatgg ttaaggtgat tctcgagttc
     1621 ttttttgagt ttaagaatcc ctctatgctc taccttgcca caggacaccg ggtgcagtgg
     1681 ctgcgctatg cagagtggct gctcacttgt cctgtcatcc ttatccgcct gagcaacctc
     1741 accggcctga gcaacgacta cagcaggaga accatgggac tccttgtctc agacatcggg
     1801 actatcgtgt ggggggctac cagcgccatg gcaaccggct atgttaaagt catcttcttt
     1861 tgtcttggat tgtgctatgg cgcgaacaca ttttttcacg ccgccaaagc atatatcgag
     1921 ggttatcata ctgtgccaaa gggtcggtgc cgccaggtcg tgaccggcat ggcatggctg
     1981 tttttcgtga gctggggtat gttcccaatt ctcttcattt tggggcccga aggttttggc
     2041 gtcctgagcg tctatggctc caccgtaggt cacacgatta ttgatctgat gagtaaaaat
     2101 tgttgggggt tgttgggaca ctacctgcgc gtcctgatcc acgagcacat attgattcac
     2161 ggagatatcc gcaaaaccac caaactgaac atcggcggaa cggagatcga ggtcgagact
     2221 ctcgtcgaag acgaagccga ggccggagcc gtgccagcgg ccgccaccat ggtgagcaag
     2281 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     2341 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     2401 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2461 ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     2521 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     2581 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2641 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2701 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     2761 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     2821 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
     2881 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2941 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagttctg ctacgagaac
     3001 gaggtgtaag gcgcgccggg ccctattcta tagtgtcacc taaatgctag agctcgctga
     3061 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3121 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3181 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3241 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3301 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3361 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3421 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3481 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3541 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3601 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3661 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3721 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3781 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3841 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3901 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3961 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4021 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4081 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4141 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4201 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4261 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4321 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4381 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4441 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4501 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4561 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4621 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4681 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4741 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4801 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4861 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4921 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4981 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5041 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5101 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5161 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5221 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5281 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5341 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5401 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5461 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5521 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5581 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5641 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5701 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5761 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5821 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5881 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5941 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6001 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6061 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6121 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6181 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6241 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6301 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6361 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6421 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6481 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6541 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6601 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6661 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6721 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6781 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6841 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6901 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6961 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7021 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7081 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7141 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7201 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7261 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7321 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7381 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7441 gcgcacattt ccccgaaaag tgccacctga cgtc
//
