LOCUS       Exported                7753 bp ds-DNA     circular SYN 02-AUG-2021
DEFINITION  Cation channelrhodopsin ChroME2f fused to mRuby2 fluorophore and 
            targeted to the neuronal soma and proximal dendrites.
ACCESSION   .
VERSION     .
KEYWORDS    pCAG-ChroME2f-mRuby2-ST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7753)
  TITLE     High performance microbial opsins for spatially and temporally 
            precise perturbations of large neuronal networks
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7753)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 2, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7753
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        67..446
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          710..1726
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1650..1672
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1734..1753
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2777..3487
                     /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          3711..3843
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3826..3845
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3852..4440
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3905..3925)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4323..4334)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4467..4484)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4473..4584
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4680..4700)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4680..4698)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4719..4735)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4837..4856)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4902..4957
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4956..4975)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5318..5334)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5342..5358
                     /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(5366..5396)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5411..5432
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5491..5686
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5537..5672
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5599..5618
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5692..5826
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5729..5748)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5783..5802
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5894..5911)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6065..6653)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6145..6164)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6824..7684)
                     /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     7447..7466
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7685..36)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
       61 tgggtcgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      121 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      181 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      241 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      301 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      361 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      421 acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc
      481 ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg
      541 cagcgatggg ggcggggggg gggggggggg cgcgcgccag gcggggcggg gcggggcgag
      601 gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc
      661 gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc
      721 gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg
      781 actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa
      841 ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg
      901 gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg
      961 cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg
     1021 cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc
     1081 gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg
     1141 gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg
     1201 agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg gcgcggggct
     1261 cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg
     1321 ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg
     1381 cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc
     1441 tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg
     1501 gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc
     1561 gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg
     1621 ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc
     1681 tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt
     1741 gctggttatt gtgctgtctc atcattttgg caaagaattc ggtaccatgg aaacagccgc
     1801 cacaatgacc cacgccttta tctcagccgt gcctagcgcc gaagccacaa ttagaggcct
     1861 gctgagcgcc gcagcagtgg tgacaccagc agcagacgct cacggagaaa cctctaacgc
     1921 cacaacagcc ggagccgatc acggttgctt cccccacatc aaccacggaa ccgagctgca
     1981 gcacaagatc gcagtgggac tccagtggtt caccgtgatc gtggctatcg tgcagctcat
     2041 cttctacggt tggcacagct tcaaggccac aaccggctgg gaggaggtct acgtctgcgt
     2101 gatcgagctc gtcaagtgct tcatcgagct gttccacgat gtcgacagcc cagccacagt
     2161 gtaccagacc aacggaggag ccgtggcttg gctgcggtac agcgagtggc tcctgacttg
     2221 ccccgtgatc ctgatccacc tgagcaacct gaccggactg cacgaagagt acagcaagcg
     2281 gaccatgacc atcctggtga ccgacatcgg caacatcgtg tgggggatca cagccgcctt
     2341 tacaaagggc cccctgaaga tcctgttctt catgatcggc ctgttctacg gcgtgacttg
     2401 cttcttccag atcgccaagg tgtatatcga gagctaccac accctgccca aaggcgtctg
     2461 ccggaagatt tgcaagatca tggcctacgt cttcttctgc tcttggctga tgttccccgt
     2521 gatgttcatc gccggacacg agggactggg cctgatcaca ccttacacca gcggaatcgg
     2581 ccacctgatc ctggatctga tcgccaagaa cacttggggc ttcctgggcc accacctgag
     2641 agtgaagatc cacgagcaca tcctgatcca cggcgacatc cggaagacaa ccaccatcaa
     2701 cgtggccggc gagaacatgg agatcgagac cttcgtcgac gaggaggagg agggaggagt
     2761 ggcggccgca ccggtaatgg tgtctaaggg cgaagagctg atcaaggaaa atatgcgtat
     2821 gaaggtggtc atggaaggtt cggtcaacgg ccaccaattc aaatgcacag gtgaaggaga
     2881 aggcaatccg tacatgggaa ctcaaaccat gaggatcaaa gtcatcgagg gaggacccct
     2941 gccatttgcc tttgacattc ttgccacgtc gttcatgtat ggcagccgta cttttatcaa
     3001 gtacccgaaa ggcattcctg atttctttaa acagtccttt cctgagggtt ttacttggga
     3061 aagagttacg agatacgaag atggtggagt cgtcaccgtc atgcaggaca ccagccttga
     3121 ggatggctgt ctcgtttacc acgtccaagt cagaggggta aactttccct ccaatggtcc
     3181 cgtgatgcag aagaagacca agggttggga gcctaataca gagatgatgt atccagcaga
     3241 tggtggtctg aggggataca ctcatatggc actgaaagtt gatggtggtg gccatctgtc
     3301 ttgctctttc gtaacaactt acaggtcaaa aaagaccgtc gggaacatca agatgcccgg
     3361 tatccatgcc gttgatcacc gcctggaaag gttagaggaa agtgacaatg aaatgttcgt
     3421 agtacaacgc gaacacgcag ttgccaagtt cgccgggctt ggtggtggga tggacgagct
     3481 gtacaaggcc ctagagcagt cccagcccat cctcaacacc aaggagatgg ccccgcagag
     3541 caagcctcca gaggagctgg agatgagcag catgcccagc cccgtggccc ctctgcccgc
     3601 acgcacggag ggcgtcatcg acatgcggag catgtccagc attgacagct tcatcagctg
     3661 tgccacggac ttccctgaag ccaccagatt ctaaggactc gaggccgcag gtaagtatca
     3721 aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac
     3781 tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca
     3841 caggtgtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     3901 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     3961 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     4021 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     4081 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     4141 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     4201 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     4261 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     4321 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     4381 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     4441 ctggaattcg agctcggtac gatcagcctc gactgtgcct tctagttgcc agccatctgt
     4501 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     4561 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     4621 tggggtgggg caggacagca agggggagga ttgggaagac aatagcccag cttttgttcc
     4681 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     4741 aattgttatc cgctaattca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg
     4801 tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta
     4861 tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt
     4921 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     4981 aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat
     5041 gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc
     5101 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
     5161 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
     5221 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag
     5281 atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5341 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5401 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5461 agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc
     5521 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5581 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     5641 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt
     5701 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5761 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5821 tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5881 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5941 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6001 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6061 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6121 ggtggcgaag cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6181 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6241 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6301 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6361 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6421 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6481 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6541 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6601 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6661 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6721 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6781 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6841 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6901 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6961 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7021 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7081 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7141 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7201 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7261 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7321 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7381 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7441 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7501 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7561 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7621 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7681 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7741 gatacatatt tga
//
