LOCUS       sequence-8322-ex        7540 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ChR2-Venus for neuronal excitation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7540)
  AUTHORS   Petreanu L, Huber D, Sobczyk A, Svoboda K
  TITLE     Channelrhodopsin-2-assisted circuit mapping of long-range callosal 
            projections.
  JOURNAL   Nat Neurosci. 2007 May . 10(5):663-8.
  PUBMED    17435752
REFERENCE   2  (bases 1 to 7540)
  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 
            Neurosci. 2007 May . 10(5):663-8."
COMMENT     Sequence Label: pCAGGS-ChR2-Venus
FEATURES             Location/Qualifiers
     source          1..7540
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             1758..2684
                     /codon_start=1
                     /product="N-terminal fragment of channelrhodopsin-2 from 
                     the alga Chlamydomonas reinhardtii (Nagel et al., 2003)"
                     /label=ChR2
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      2712..2721
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2718..2720,2721..2723,2724..3437)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2763..2784)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3024..3043)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3371..3392
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          3463..3596
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3579..3598
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3605..4193
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3658..3678)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4076..4087)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4220..4237)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4226..4337
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4564..4583)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4629..4684
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4683..4702)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5043..5059)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5056..5078)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5067..5083
                     /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(5091..5097,5098..5115,5116..5121))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5136..5157
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5216..5411
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5262..5397
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5417..5551
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5454..5473)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5508..5527
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5619..5636)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5790..6378)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5870..5889)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6549..7340,7341..7409))
                     /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     7172..7191
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7410..7514)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg ggcgagggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tgttgtgctg tctcatcatt ttggcaaaga attcggtacc gcgggcccgg
     1741 gatccaccgg tgctagcatg gattatggag gcgccctgag tgccgttggg cgcgagctgc
     1801 tatttgtaac gaacccagta gtcgtcaatg gctctgtact tgtgcctgag gaccagtgtt
     1861 actgcgcggg ctggattgag tcgcgtggca caaacggtgc ccaaacggcg tcgaacgtgc
     1921 tgcaatggct tgctgctggc ttctccatcc tactgcttat gttttacgcc taccaaacat
     1981 ggaagtcaac ctgcggctgg gaggagatct atgtgtgcgc tatcgagatg gtcaaggtga
     2041 ttctcgagtt cttcttcgag tttaagaacc cgtccatgct gtatctagcc acaggccacc
     2101 gcgtccagtg gttgcgttac gccgagtggc ttctcacctg cccggtcatt ctcattcacc
     2161 tgtcaaacct gacgggcttg tccaacgact acagcaggcg caccatgggt ctgcttgtgt
     2221 ctgatattgg cacaattgtg tggggcgcca cttccgccat ggccaccgga tacgtcaagg
     2281 tcatcttctt ctgcctgggt ctgtgttatg gtgctaacac gttctttcac gctgccaagg
     2341 cctacatcga gggttaccac accgtgccga agggccggtg tcgccaggtg gtgactggca
     2401 tggcttggct cttcttcgta tcatggggta tgttccccat cctgttcatc ctcggccccg
     2461 agggcttcgg cgtcctgagc gtgtacggct ccaccgtcgg ccacaccatc attgacctga
     2521 tgtcgaagaa ctgctggggt ctgctcggcc actacctgcg cgtgctgatc cacgagcata
     2581 tcctcatcca cggcgacatt cgcaagacca ccaaattgaa cattggtggc actgagattg
     2641 aggtcgagac gctggtggag gacgaggccg aggctggcgc ggtcaacaag ggcaccggca
     2701 agccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     2761 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     2821 agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc
     2881 ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cctgcagtgc ttcgcccgct
     2941 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     3001 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     3061 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     3121 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg
     3181 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg
     3241 gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     3301 tgctgcccga caaccactac ctgagctacc agtccaagct gagcaaagac cccaacgaga
     3361 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     3421 acgagctgta caagtaaagc ggccccggac tcgaggccgc aggtaagtat caaggttaca
     3481 agacaggttt aaggagacca catagaaact gggcttgtcg agacagagaa gactcttgcg
     3541 tttctgatag gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt
     3601 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3661 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3721 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     3781 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     3841 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     3901 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     3961 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     4021 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     4081 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     4141 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat
     4201 tcgagctcgg tacgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     4261 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     4321 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     4381 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggccagcttt tgttcccttt
     4441 agggttaatt gcgcgcttgg cgctaatcat ggtcatagct gtttccactc ctcaggtgca
     4501 ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg
     4561 agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac
     4621 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     4681 ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt
     4741 ttagagtttg gcaacatatg ccatatgctg gctgccatga acaaaggtgg ctataaagag
     4801 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga
     4861 cttgaggtta gatttttttt tatattttgt ttgtgttatt tttttcttta acatccctaa
     4921 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca
     4981 tagctgtccc tcttctctta tgaagatccc tcgacctgca gcccaagctt ggcgtaatca
     5041 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5101 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5161 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc
     5221 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     5281 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5341 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5401 gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5461 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5521 actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg
     5581 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5641 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5701 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5761 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5821 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5881 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5941 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     6001 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6061 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6121 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6181 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     6241 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6301 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6361 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6421 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6481 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6541 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6601 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6661 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6721 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6781 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6841 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6901 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6961 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7021 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7081 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7141 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7201 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7261 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7321 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7381 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7441 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7501 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
