LOCUS       sequence-220130-        6499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses channelrhodopsin 2 fused with EYFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6499)
  AUTHORS   Messier JE, Chen H, Cai ZL, Xue M
  TITLE     Targeting light-gated chloride channels to neuronal somatodendritic 
            domain reduces their excitatory effect in the axon.
  JOURNAL   Elife. 2018 Aug 9;7. pii: 38506. doi: 10.7554/eLife.38506.
  PUBMED    30091701
REFERENCE   2  (bases 1 to 6499)
  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: "Elife."; 
            date: "2018-08-9"; volume: "7. pii: 38506. doi"; pages: " 
            10.7554/eLife.38506"
COMMENT     Sequence Label: pCAG-hChR2(H134R)-EYFP
FEATURES             Location/Qualifiers
     source          1..6499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        8..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..665
                     /label=chicken beta-actin promoter
     intron          666..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1598..1620
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1698..1717
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1765..1774
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1771..2697
                     /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      2707..2716
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2713..2715,2716..2718,2719..3432)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2758..2779)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3019..3038)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3366..3387
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3521..3540)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3586..3641
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3640..3659)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4002..4018)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4002..4018)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4015..4037)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4026..4042
                     /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(4050..4056,4057..4074,4075..4080))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4095..4116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4175..4370
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4221..4356
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4376..4510
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4413..4432)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4467..4486
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4578..4595)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4749..5337)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4829..4848)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5508..6299,6300..6368))
                     /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     6131..6150
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6369..6473)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgtcgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gactatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatgggt cgaggtgagc cccacgttct gcttcactct
      421 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      481 tgcagcgatg ggggcggggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg
      541 ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
      601 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
      661 gggcgggagt cgctgcgttg ccttcgcccc gtgccccgct ccgcgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggctcgt ttcttttctg tggctgcgtg
      841 aaagccttaa agggctccgg gagggccctt tgtgcggggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggcccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcgtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcggcggtcg ggctgtaacc cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtgcggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctggcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcgggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg
     1501 ccttcgtgcg tcgccgcgcc gccgtcccct tctccatctc cagcctcggg gctgccgcag
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga
     1741 attctgcagt cgacggtacc gctagccacc atggactatg gcggcgcttt gtctgccgtc
     1801 ggacgcgaac ttttgttcgt tactaatcct gtggtggtga acgggtccgt cctggtccct
     1861 gaggatcaat gttactgtgc cggatggatt gaatctcgcg gcacgaacgg cgctcagacc
     1921 gcgtcaaatg tcctgcagtg gcttgcagca ggattcagca ttttgctgct gatgttctat
     1981 gcctaccaaa cctggaaatc tacatgcggc tgggaggaga tctatgtgtg cgccattgaa
     2041 atggttaagg tgattctcga gttctttttt gagtttaaga atccctctat gctctacctt
     2101 gccacaggac accgggtgca gtggctgcgc tatgcagagt ggctgctcac ttgtcctgtc
     2161 atccttatcc gcctgagcaa cctcaccggc ctgagcaacg actacagcag gagaaccatg
     2221 ggactccttg tctcagacat cgggactatc gtgtgggggg ctaccagcgc catggcaacc
     2281 ggctatgtta aagtcatctt cttttgtctt ggattgtgct atggcgcgaa cacatttttt
     2341 cacgccgcca aagcatatat cgagggttat catactgtgc caaagggtcg gtgccgccag
     2401 gtcgtgaccg gcatggcatg gctgtttttc gtgagctggg gtatgttccc aattctcttc
     2461 attttggggc ccgaaggttt tggcgtcctg agcgtctatg gctccaccgt aggtcacacg
     2521 attattgatc tgatgagtaa aaattgttgg gggttgttgg gacactacct gcgcgtcctg
     2581 atccacgagc acatattgat tcacggagat atccgcaaaa ccaccaaact gaacatcggc
     2641 ggaacggaga tcgaggtcga gactctcgtc gaagacgaag ccgaggccgg agccgtgcca
     2701 gcggccgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2761 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     2821 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2881 ccctggccca ccctcgtgac caccttcggc tacggcctgc agtgcttcgc ccgctacccc
     2941 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3001 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3061 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3121 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     3181 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     3241 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     3301 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     3361 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3421 ctgtacaagt aatagggcgc gccactcctc aggtgcaggc tgcctatcag aaggtggtgg
     3481 ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa
     3541 attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta
     3601 ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga
     3661 gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc
     3721 atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc
     3781 cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta
     3841 tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac
     3901 tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat
     3961 ggagatccct cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4021 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4081 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4141 ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca gcaaccatag
     4201 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4261 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     4321 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctaactt
     4381 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4441 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4501 tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4561 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4621 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4681 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4741 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4801 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4861 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4921 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4981 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5041 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5101 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5161 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5221 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5281 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5341 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5401 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5461 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5521 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5581 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5641 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5701 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5761 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5821 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5881 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5941 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6001 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6061 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6121 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6181 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6241 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6301 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6361 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6421 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6481 ccccgaaaag tgccacctg
//
