LOCUS       Exported                7814 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Artificial anion channelrhodopsin iC++ fused to ER export signal and
            fluorophore mRuby2 and targeted to the neuronal soma and proximal 
            dendrites.
ACCESSION   .
VERSION     .
KEYWORDS    pCAG-eiC++-mRuby2-ST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7814)
  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 7814)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7814
                     /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             2758..3468
                     /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          3710..3842
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3825..3844
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3851..4439
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3904..3924)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4322..4333)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4466..4483)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4472..4583
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4679..4699)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4679..4697)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4718..4734)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4836..4855)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4901..4956
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4955..4974)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5317..5333)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     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(5365..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."
     promoter        5490..5685
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5536..5671
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5598..5617
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5691..5825
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5728..5747)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5782..5801
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5893..5910)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6064..6652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6144..6163)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6823..7683)
                     /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     7446..7465
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7684..7788)
                     /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 gccagggggg gggggggggg gggggggggg
      541 ggggggggcg gggcggagag 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 tcggtacccc
     1741 accaccatgg actatggcgg cgctttgtct gccgtcggac tcttccaaac ctcatacact
     1801 cttgagaaca atggttctgt tatttgcatc ccgaacaacg gccagtgctt ctgcttggct
     1861 tggcttaaat ccaacggaac aaatgccgag aagttggctg ccaacattct gcagtggatt
     1921 tcttttgcgc tttcagcgct ctgcctgatg ttctacggct accagacctg gaagtctact
     1981 tgcggctggg agaatattta cgtggccacg atccagatga tcaagttcat catcgagtat
     2041 ttccatagct ttgacgaacc tgcggtgatc tactcatcca acggcaacaa gaccaggtgg
     2101 cttcgttacg cgagctggct gctgacctgc cctgtcattc ttatccacct gagcaacctt
     2161 acgggtctgg cgaacgacta taacaagcgt accatgggtc tgctggtgtc agatatcggc
     2221 acgatcgtgt ggggcaccac ggccgcgctg tccaagggat acgtccgtgt cattttcttc
     2281 ctgatgggcc tgtgctacgg catctacaca ttcttcaacg cagccaaggt ctacattgag
     2341 gcgtaccaca ccgtgccgaa gggccggtgt cgccaggtgg tgactggcat ggcttggctc
     2401 ttcttcgtat catggggtat gttccccatc ctgttcatcc tcggccccga gggcttcggc
     2461 gtcctgagca ggtacggctc caacgtcggc cacaccatca ttgacctgat gtcgaagcag
     2521 tgctggggtc tgctcggcca ctacctgcgc gtgctgatcc acagccatat cctcatccac
     2581 ggcgacattc gcaagaccac caaattgaac attggtggca ctgagattga ggtcgagacg
     2641 ctggtggagg acgaggccga ggctggcgcg gtcgcggccg ccaagagcag gatcaccagc
     2701 gagggcgagt acatccccct ggaccagatc gacatcaacg tggcggcggc accggtaatg
     2761 gtgtctaagg gcgaagagct gatcaaggaa aatatgcgta tgaaggtggt catggaaggt
     2821 tcggtcaacg gccaccaatt caaatgcaca ggtgaaggag aaggcaatcc gtacatggga
     2881 actcaaacca tgaggatcaa agtcatcgag ggaggacccc tgccatttgc ctttgacatt
     2941 cttgccacgt cgttcatgta tggcagccgt acttttatca agtacccgaa aggcattcct
     3001 gatttcttta aacagtcctt tcctgagggt tttacttggg aaagagttac gagatacgaa
     3061 gatggtggag tcgtcaccgt catgcaggac accagccttg aggatggctg tctcgtttac
     3121 cacgtccaag tcagaggggt aaactttccc tccaatggtc ccgtgatgca gaagaagacc
     3181 aagggttggg agcctaatac agagatgatg tatccagcag atggtggtct gaggggatac
     3241 actcatatgg cactgaaagt tgatggtggt ggccatctgt cttgctcttt cgtaacaact
     3301 tacaggtcaa aaaagaccgt cgggaacatc aagatgcccg gtatccatgc cgttgatcac
     3361 cgcctggaaa ggttagagga aagtgacaat gaaatgttcg tagtacaacg cgaacacgca
     3421 gttgccaagt tcgccgggct tggtggtggg atggacgagc tgtacaaggc cctagagcag
     3481 tcccagccca tcctcaacac caaggagatg gccccgcaga gcaagcctcc agaggagctg
     3541 gagatgagca gcatgcccag ccccgtggcc cctctgcccg cacgcacgga gggcgtcatc
     3601 gacatgcgga gcatgtccag cattgacagc ttcatcagct gtgccacgga cttccctgaa
     3661 gccaccagat tcttctgcta cgagaacgag gtgtaactcg aggccgcagg taagtatcaa
     3721 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
     3781 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     3841 aggtgtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     3901 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     3961 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     4021 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     4081 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     4141 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     4201 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     4261 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     4321 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     4381 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     4441 tggaattcga gctcggtacg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     4501 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     4561 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     4621 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcccagc ttttgttccc
     4681 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4741 attgttatcc gctaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
     4801 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
     4861 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     4921 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     4981 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     5041 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     5101 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     5161 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     5221 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     5281 tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5341 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5401 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5461 gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg
     5521 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     5581 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     5641 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta
     5701 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5761 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5821 ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5881 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5941 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6001 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6061 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6121 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6181 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6241 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6301 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6361 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6421 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6481 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6541 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6601 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6661 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6721 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6781 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6841 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6901 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6961 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7021 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7081 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7141 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7201 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7261 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7321 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7381 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7441 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7501 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7561 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7621 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7681 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7741 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7801 aaaagtgcca cctg
//
