LOCUS       Exported                7764 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Cation channelrhodopsin ChETA fused to mRuby2 fluorophore and 
            targeted to the neuronal soma and proximal dendrites.
ACCESSION   .
VERSION     .
KEYWORDS    pCAG-ChETA-mRuby2-ST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7764)
  AUTHORS   Mardinly AR, Oldenburg IA, Pegard NC, Sridharan S, Lyall EH, Chesnov
            K, Brohawn SG, Waller L, Adesnik H
  TITLE     Precise multimodal optical control of neural ensemble activity.
  JOURNAL   Nat Neurosci. 2018 Apr 30. pii: 10.1038/s41593-018-0139-8. doi: 
            10.1038/s41593-018-0139-8.
  PUBMED    29713079
REFERENCE   2  (bases 1 to 7764)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7764
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1766..2692
                     /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
                     PSMLYLATGHRVQWLRYATWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             2729..3439
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     intron          3660..3792
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3775..3794
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3801..4389
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3854..3874)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4272..4283)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4416..4433)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4422..4533
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4629..4649)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4629..4647)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4668..4684)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4786..4805)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4851..4906
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4905..4924)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5267..5283)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5291..5307
                     /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(5315..5345)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5360..5381
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5440..5635
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5486..5621
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5548..5567
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5641..5775
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5678..5697)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5732..5751
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5843..5860)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6014..6602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6094..6113)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6773..7633)
                     /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     7396..7415
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7634..7738)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gggggggggg gggggggggg
      541 ggggggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcggtaccgc
     1741 gggcccggga tccaccggtg ctagcatgga ttatggaggc gccctgagtg ccgttgggcg
     1801 cgagctgcta tttgtaacga acccagtagt cgtcaatggc tctgtacttg tgcctgagga
     1861 ccagtgttac tgcgcgggct ggattgagtc gcgtggcaca aacggtgccc aaacggcgtc
     1921 gaacgtgctg caatggcttg ctgctggctt ctccatccta ctgcttatgt tttacgccta
     1981 ccaaacatgg aagtcaacct gcggctggga ggagatctat gtgtgcgcta tcgagatggt
     2041 caaggtgatt ctcgagttct tcttcgagtt taagaacccg tccatgctgt atctagccac
     2101 aggccaccgc gtccagtggt tgcgttacgc cacgtggctt ctcacctgcc cggtcattct
     2161 cattcacctg tcaaacctga cgggcttgtc caacgactac agcaggcgca ccatgggtct
     2221 gcttgtgtct gatattggca caattgtgtg gggcgccact tccgccatgg ccaccggata
     2281 cgtcaaggtc atcttcttct gcctgggtct gtgttatggt gctaacacgt tctttcacgc
     2341 tgccaaggcc tacatcgagg gttaccacac cgtgccgaag ggccggtgtc gccaggtggt
     2401 gactggcatg gcttggctct tcttcgtatc atggggtatg ttccccatcc tgttcatcct
     2461 cggccccgag ggcttcggcg tcctgagcgt gtacggctcc accgtcggcc acaccatcat
     2521 tgacctgatg tcgaagaact gctggggtct gctcggccac tacctgcgcg tgctgatcca
     2581 cgagcatatc ctcatccacg gcgacattcg caagaccacc aaattgaaca ttggtggcac
     2641 tgagattgag gtcgagacgc tggtggagga cgaggccgag gctggcgcgg tcaacaaggg
     2701 caccggcaag ccaccggtcg caccggtaat ggtgtctaag ggcgaagagc tgatcaagga
     2761 aaatatgcgt atgaaggtgg tcatggaagg ttcggtcaac ggccaccaat tcaaatgcac
     2821 aggtgaagga gaaggcaatc cgtacatggg aactcaaacc atgaggatca aagtcatcga
     2881 gggaggaccc ctgccatttg cctttgacat tcttgccacg tcgttcatgt atggcagccg
     2941 tacttttatc aagtacccga aaggcattcc tgatttcttt aaacagtcct ttcctgaggg
     3001 ttttacttgg gaaagagtta cgagatacga agatggtgga gtcgtcaccg tcatgcagga
     3061 caccagcctt gaggatggct gtctcgttta ccacgtccaa gtcagagggg taaactttcc
     3121 ctccaatggt cccgtgatgc agaagaagac caagggttgg gagcctaata cagagatgat
     3181 gtatccagca gatggtggtc tgaggggata cactcatatg gcactgaaag ttgatggtgg
     3241 tggccatctg tcttgctctt tcgtaacaac ttacaggtca aaaaagaccg tcgggaacat
     3301 caagatgccc ggtatccatg ccgttgatca ccgcctggaa aggttagagg aaagtgacaa
     3361 tgaaatgttc gtagtacaac gcgaacacgc agttgccaag ttcgccgggc ttggtggtgg
     3421 gatggacgag ctgtacaagg ccctagagca gtcccagccc atcctcaaca ccaaggagat
     3481 ggccccgcag agcaagcctc cagaggagct ggagatgagc agcatgccca gccccgtggc
     3541 ccctctgccc gcacgcacgg agggcgtcat cgacatgcgg agcatgtcca gcattgacag
     3601 cttcatcagc tgtgccacgg acttccctga agccaccaga ttctaactcg aggccgcagg
     3661 taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac
     3721 agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct
     3781 ttctctccac aggtgtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     3841 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     3901 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     3961 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     4021 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     4081 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     4141 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga
     4201 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     4261 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     4321 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     4381 cctccccgcc tggaattcga gctcggtacg atcagcctcg actgtgcctt ctagttgcca
     4441 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     4501 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     4561 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcccagc
     4621 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     4681 cctgtgtgaa attgttatcc gctaattcac tcctcaggtg caggctgcct atcagaaggt
     4741 ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc
     4801 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
     4861 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     4921 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
     4981 tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa
     5041 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
     5101 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
     5161 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
     5221 cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc
     5281 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5341 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5401 ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac
     5461 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     5521 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     5581 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     5641 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5701 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5761 tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5821 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5881 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5941 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6001 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6061 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6121 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6181 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6241 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6301 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6361 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6421 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6481 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6541 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6601 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6661 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6721 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6781 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6841 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6901 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6961 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7021 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7081 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7141 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7201 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7261 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7321 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7381 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7441 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7501 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7561 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7621 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7681 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7741 catttccccg aaaagtgcca cctg
//
