LOCUS       Exported                7693 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Anion channelrhodopsin GtACR1 fused to ER export signal, mRuby2 and 
            targeted to the neuronal soma and proximal dendrites.
ACCESSION   .
VERSION     .
KEYWORDS    pCAG-eGtACR1-mRuby2-ST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7693)
  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 7693)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7693
                     /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             2637..3347
                     /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          3589..3721
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3704..3723
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3730..4318
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3783..3803)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4201..4212)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4345..4362)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4351..4462
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4558..4578)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4558..4576)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4597..4613)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4715..4734)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4780..4835
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4834..4853)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5196..5212)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5220..5236
                     /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(5244..5274)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5289..5310
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5369..5564
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5415..5550
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5477..5496
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5570..5704
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5607..5626)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5661..5680
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5772..5789)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5943..6531)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6023..6042)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6702..7562)
                     /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     7325..7344
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7563..7667)
                     /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 gcgggggggg gggggggggg
      541 gggggggggg rggcggagag 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 tcggtaccct
     1741 ccatgagcag catcacctgt gatcccgcca tctacggcga atggagcagg gagaaccagt
     1801 tctgcgtgga gaagagcctg atcaccctgg acggcatcaa gtacgtccag ctggtgatgg
     1861 ccgtcgtgag cgcctgtcag gtgttcttca tggtgaccag agcccccaag gtgccctggg
     1921 aagccatcta cctgcccacc accgagatga tcacctattc cctggccttc accggaaacg
     1981 gctacatcag agtggccaat ggcaagtacc tgccctgggc cagaatggcc agctggctgt
     2041 gcacctgccc tatcatgctg ggcctggtga gcaatatggc cctcgtgaag tacaagagca
     2101 tccccctcaa ccctatgatg atcgccgcct ccagcatctg caccgtgttc ggcatcaccg
     2161 cctccgtggt gctagacccg ctgcacgtgt ggctgtactg tttcatcagc agcatcttct
     2221 tcatcttcga gatggtggtg gccttcgcca ttttcgccat taccatccac gatttccaga
     2281 ccatcggctc ccccatgtcc ctgaaggtgg tggagaggct gaagctgatg aggatcgtgt
     2341 tctacgtgag ctggatggcc taccctatcc tgtggagctt ctccagcacc ggcgcctgca
     2401 tcatgagcga gaacaccagc agcgtgctgt acctgctggg cgacgctctg tgcaagaaca
     2461 cctacggcat cctgctgtgg gctacaacct ggggcctgct gaacggcaag tgggacaggg
     2521 attacgtgaa gggcaggaac gtggacggca ccctgatgcc tgagtacgag caggacctgg
     2581 agaagggcaa caccgagagg tacgaggacg ccagagccgg cgagaccgca ccggtaatgg
     2641 tgtctaaggg cgaagagctg atcaaggaaa atatgcgtat gaaggtggtc atggaaggtt
     2701 cggtcaacgg ccaccaattc aaatgcacag gtgaaggaga aggcaatccg tacatgggaa
     2761 ctcaaaccat gaggatcaaa gtcatcgagg gaggacccct gccatttgcc tttgacattc
     2821 ttgccacgtc gttcatgtat ggcagccgta cttttatcaa gtacccgaaa ggcattcctg
     2881 atttctttaa acagtccttt cctgagggtt ttacttggga aagagttacg agatacgaag
     2941 atggtggagt cgtcaccgtc atgcaggaca ccagccttga ggatggctgt ctcgtttacc
     3001 acgtccaagt cagaggggta aactttccct ccaatggtcc cgtgatgcag aagaagacca
     3061 agggttggga gcctaataca gagatgatgt atccagcaga tggtggtctg aggggataca
     3121 ctcatatggc actgaaagtt gatggtggtg gccatctgtc ttgctctttc gtaacaactt
     3181 acaggtcaaa aaagaccgtc gggaacatca agatgcccgg tatccatgcc gttgatcacc
     3241 gcctggaaag gttagaggaa agtgacaatg aaatgttcgt agtacaacgc gaacacgcag
     3301 ttgccaagtt cgccgggctt ggtggtggga tggacgagct gtacaaggcc ctagagcagt
     3361 cccagcccat cctcaacacc aaggagatgg ccccgcagag caagcctcca gaggagctgg
     3421 agatgagcag catgcccagc cccgtggccc ctctgcccgc acgcacggag ggcgtcatcg
     3481 acatgcggag catgtccagc attgacagct tcatcagctg tgccacggac ttccctgaag
     3541 ccaccagatt cttctgctac gagaacgagg tgtaactcga ggccgcaggt aagtatcaag
     3601 gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc
     3661 ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca
     3721 ggtgtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     3781 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     3841 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     3901 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     3961 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     4021 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     4081 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     4141 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     4201 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     4261 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     4321 ggaattcgag ctcggtacga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     4381 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     4441 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     4501 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcccagct tttgttccct
     4561 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4621 ttgttatccg ctaattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg
     4681 tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg
     4741 gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca
     4801 ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa
     4861 atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc
     4921 tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg
     4981 ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt
     5041 gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca
     5101 gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat
     5161 ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5221 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5281 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5341 tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc atagtcccgc
     5401 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5461 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     5521 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta acttgtttat
     5581 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5641 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5701 gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5761 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5821 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5881 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5941 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6001 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6061 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6121 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6181 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6241 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6301 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6361 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6421 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6481 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6541 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6601 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6661 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6721 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6781 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6841 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6901 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6961 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7021 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7081 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7141 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7201 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7261 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7321 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7381 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7441 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7501 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7561 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7621 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7681 aaagtgccac ctg
//
